Dr. Alice Bertaina completed her MD degree at the University of Pavia in Italy, her fellowship in hematopoietic stem cell transplantation (HSCT) at the Bambino Gesù Children’s Hospital in Rome, and her PhD degree in Immunology and Biotechnology at Tor Vergata University in Rome. Until joining Stanford University in 2017, she was Head of the Stem Cell Transplant Unit in the Department of Hematology and Oncology at the Bambino Gesù Children’s Hospital in Rome (this institution currently has the largest number of children transplanted with hematopoietic progenitors/stem cells in Europe).

Dr. Bertaina is an expert in the field of allogeneic HSCT in pediatric patients affected by hematological malignancies or nonmalignant disorders. In particular, she has pioneered the novel approach of graft manipulation based on the physical elimination of alfa/beta T cells and B cells. Dr. Bertaina has excellent clinical and biological expertise, as demonstrated by her publications in the field of pediatric hematology and oncology. Moreover, she is expert in different aspects of immunological reconstitution of children given an allograft of hematopoietic stem cell, paying particular attention to innate immunity.

Clinical Focus

  • Pediatric Stem Cell Transplantation
  • Graft engineering
  • Pediatric Hematology-Oncology

Administrative Appointments

  • Co-Director, Bass Center for Childhood Cancer and Blood Diseaeses, Lucile Packard Children's Hospital, Stanford University, CA, USA (2020 - Present)
  • Section Chief, Stem Cell Transplant and Regenerative Medicine, Lucile Packard Children's Hospital, Stanford University, CA, USA (2020 - Present)
  • Medical Director of Inpatient Services, Lucile Packard Children's Hospital, Stanford University, CA, USA (2019 - Present)
  • Head of Stem Cell Transplant Unit, Department of Hematology and Oncology, Bambino Gesù Children’s Hospital Rome, Italy (2013 - 2017)
  • Physician Assistant and Researcher, Department of Hematology and Oncology, Bambino Gesù Children’s Hospital, Rome, Italy (2010 - 2013)
  • Residency in Pediatrics, Pediatric Hematology and Oncology, Fondazione IRCCS Policlinico San Matteo, Pavia, Italy (2008 - 2010)
  • Medical and Research Fellowship, Department of Pediatrics, Fondazione IRCCS Policlinico San Matteo, Pavia, Italy (2004 - 2008)

Honors & Awards

  • Lorry Lokey Faculty Scholar, Stanford University School of Medicine (2019-)
  • Press release 'alfa/beta T-cell depleted haplo-transplant in children with acute leukemia', 55th ASH Meeting, Engineering Cell Therapies Press Program, New Orleans (December 2013)
  • Abstract Travel Award 'Identification of deregulated microRNAs in JMML', 2016 International JMML Symposium, San Diego (December 2016)
  • Invited to participate in the Annual National Ceremony for Cancer Research, Palazzo del Quirinale, By Invitation from the President of the Italian Republic (October 2015)

Boards, Advisory Committees, Professional Organizations

  • Member, ISCT Stem Cell Engineering Committee (2020 - Present)
  • Member, Child Health Research Institute Stanford (CHRI) (2017 - Present)
  • Member, Italian Group for Bone Marrow Transplantation (2017 - Present)
  • Chair, Paediatric transplantation Session, EBMT 43 rd Annual Meeting, Marseille, France, March 26-29, 2017 (2017 - 2017)
  • Member, GITMO project entitled ‘3 Steps’, aiming at defining strategies of prophylaxis and treatment of veno-occlusive disease in transplant recipients (2016 - Present)
  • Member, Inborn Errors Working Group, EBMT. (2016 - Present)
  • Peer Reviewed Journal Reviewer, Blood Clinical Cancer Research Frontiers in Immunology BBMT BMT Oncoimmunology Haematologica (2016 - Present)
  • Peer Reviewed Journal Reviewer, Bone Marrow Transplantation (2015 - Present)
  • Head, HSCT Working Party Group of AIEOP (Italian Association of Pediatric Hematology and Oncology) (2015 - 2017)
  • Board Member, EBMT PDWP (European Society for Blood and Marrow Transplantation Pediatric Disease Working Party) (2014 - 2017)
  • Member, American Society of Hematology (2013 - Present)
  • Member, Comitato per il Controllo delle Infezioni Ospedalerie (CCIO) at the OPBG, Rome (2011 - 2017)
  • Member, Associazione Italiana di Ematologia ed Oncaologia Pediatrics (2010 - Present)
  • Member, European Society for Blood and Marrow Transplantation (2010 - Present)

Professional Education

  • Board Certification: Ordine dei Medici Chirurghi e degli Odontoiatri della provincial di Pavia, Internal Medicine (2006)
  • Fellowship: Ospedale Bambino Gesu' (2013) Italy
  • Residency: University of Pavia (2010) Italy
  • Medical Education: University of Pavia (2005) Italy
  • PhD, Tor Vergata University, Immunology and Biotechnology (2013)
  • Residency in Pediatrics, University of Pavia, Pavia, Italy, Thesis title: “NK-alloreactivity and outcome of pediatric patients with acute leukemia given an HLA-haploidentical stem cell transplantation” under Prof. Franco Locatelli supervision (2010)
  • Post-graduate fellowship, Bambino Gesù Children’s Hospital, Rome, Italy, Hematopoietic Stem Cell Transplantation (basic and applied research) (2010)
  • Medical Degree, University of Pavia, Pavia, Italy, Thesis title: “Human Metapneumovirus II in acute pediatric respiratory infection: epidemiology, clinical picture and diagnosis” (2005)

Current Research and Scholarly Interests

Dr. Bertaina is a highly experienced clinician and will play a key role in supporting Section Chief Dr. Rajni Agarwal and Clinical Staff in the Stem Cell Transplant Unit at Lucile Packard Children’s Hospital. She will also continue her research on immune recovery and miRNA, understanding the mechanisms underlying immune reconstitution, Graft-versus-Host Disease (GvHD), and leukemia relapse after allogeneic HSCT in pediatric patients affected by hematological malignant and non-malignant disorders.

Clinical Trials

  • Early Trial of Allogeneic Hematopoietic Stem Cell Transplantation for Patients Who Will Receive a Kidney Transplant From the Same Donor Recruiting

    This is a single center, non-randomized, non-controlled open-label phase 1b/2a trial of performing sequential αβdepleted-HSCT and KT in patients requiring KT to prevent kidney rejection post-KT, in the absence of any post-KT immunosuppression, to abrogate the need for lifelong immunosuppression, the risk of chronic rejection and, ultimately, the need for repeated transplantation.

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  • Stem Cell Transplant From Donors After Alpha Beta Cell Depletion in Children and Adults With T-allo10 Cells Addback Recruiting

    The purpose of this study is to determine the safety of a cell therapy, T-allo10, after αβdepleted-HSCT in the hopes that it will boost the adaptive immune reconstitution of the patient while sparing the risk of developing severe Graft-versus-Host Disease (GvHD). The primary objective of Phase 1a is to determine the recommended Phase 2 dose (RP2D) administered after infusion of αβdepleted-HSCT in children and young adults with hematologic malignancies. A Phase 1b extension will occur after dose escalation, enrolling at the RP2D for the T-allo10 cells determined in the Phase 1 portion to evaluate the safety and efficacy of infusion of T-allo10 after receipt of αβdepleted-HSCT. Additionally, Phase 1b aims to explore improvements in immune reconstitution. All participants on this study must be enrolled on another study: NCT04249830

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  • Stem Cell Transplant From Donors After Alpha Beta Cell Depletion in Children and Young Adults Recruiting

    The purpose of the CliniMACS® TCRαβ-Biotin System and CliniMACS® CD19 is to improve the safety and efficacy of allogeneic HLA-partially matched related or unrelated donors HSCT when no matched donors are available, to treat malignant and nonmalignant disorders for which HSCT is the recommended best available therapy. Initially this device will be used in a single-center, open-label, single-arm, phase II clinical trial to evaluate the efficacy of haploidentical PBSC grafts depleted of TCRα/β+ and CD19+ cells using the CliniMACS® TCRαβ/CD19 System in children and adults with hematological and non-hematological malignancies.

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  • KIR Favorable Mismatched Haplo Transplant and KIR Polymorphism in ALL/AML/MDS Allo-HCT Children Not Recruiting

    This is a phase II, open-label, non-randomized, prospective study of haploidentical transplantation using KIR-favorable donors for children with acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML) and myelodysplastic syndrome (MDS) undergoing allogeneic hematopoietic cell transplantation (HCT). The relationship of KIR2DL1 polymorphisms to survival in children with these diseases undergoing any approach to allogeneic HCT during the study time frame will also be determined.

    Stanford is currently not accepting patients for this trial. For more information, please contact Cancer Clinical Trials Office (CCTO), 650-498-7061.

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  • Safety Study of Gene Modified Donor T-cells Following TCRαβ+ Depleted Stem Cell Transplant Not Recruiting

    This study will evaluate pediatric patients with malignant or non-malignant blood cell disorders who are having a blood stem cell transplant depleted of T cell receptor (TCR) alfa and beta cells that comes from a partially matched family donor. The study will assess whether immune cells, called T cells, from the family donor, that are specially grown in the laboratory and given back to the patient along with the stem cell transplant can help the immune system recover faster after transplant. As a safety measure these T cells have been programmed with a self-destruct switch so that they can be destroyed if they start to react against tissues (Graft versus host disease).

    Stanford is currently not accepting patients for this trial. For more information, please contact Cancer Clinical Trials Office (CCTO), 650-498-7061.

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  • Study of GDX012 in Patients With MRD Positive AML Not Recruiting

    The purpose of this first-in-human study is to assess the safety, tolerability, antileukemic activity and maximum tolerated dose (MTD) of GDX012 in AML patients who are MRD positive by multiparametric flow cytometry. The study will consist of a dose escalation stage to evaluate various doses of GDX012 after a lymphodepletion regimen comprising fludarabine and cyclophosphamide. Following determination of the MTD of GDX012, the study will expand at the MTD. Patients will be followed up for 12 months, after receiving GDX012.

    Stanford is currently not accepting patients for this trial. For more information, please contact Cancer Clinical Trials Office (CCTO), 650-498-7061.

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  • US Phase I Study of ECT-001-CB in Patients With Sickle-Cell Disease Not Recruiting

    The application of experimental hematopoietic cell transplantation (HCT) therapy in sickle-cell disease (SCD) must strike a balance between the underlying disease severity and the possibility of a direct benefit of the treatment, particularly in pediatric populations. Clinical studies in adults with SCD have focused on interventions that prolong survival and improve the quality of life. Unlike children, adults with SCD are much more likely to have a debilitating complication. As a result, the risk/benefit ratio of HCT is very favorable in adults, particularly if an approach to HCT that defines an acceptable level of toxicity can be established. Whereas hematopoietic stem cell transplantation (HSCT) remains the only curative treatment currently available for patients with SCD, the morbidity, the frequent irreversible damage in target organs and the mortality reported in the natural course of patients with severe SCD are strong incentives to perform HSCTs in younger age groups. For those who lack a matched related donor, CB transplant is an appealing option, but despite been less problematic, CB accessibility related to cell dose of appropriately matched cord blood unit (CBU) remains a significant issue. Through a 7-day culture process of a CBU's hematopoietic stem cell HSCs with the UM171 compound, the total cell dose is increased mitigating this limitation. UM171-CB expansion (ECT-001-CB) allows a greater CB accessibility, the selection of better matched cords that might translate into favourable clinical outcomes as reported in previous trials, including a lower risk of graft-versus-host disease. After CB selection and ex-vivo expansion, ECT-001-CB transplant will follow a myeloablative reduced-toxicity conditioning regimen consisting of rATG, busulfan and fludarabine with doses of all agents optimized to the individual using model-based dosing and will be followed by standard supportive care and GVHD prophylaxis consisting of tacrolimus and MMF.

    Stanford is currently not accepting patients for this trial.

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2023-24 Courses

Stanford Advisees

All Publications

  • Model based ATG in αβhaplo-HSCT facilitates engraftment, expedites T-cell recovery, and mitigates the risk of acute GvHD. Transplantation and cellular therapy Giulia, B., Lyndsie, H., Linda, O., Gopin, S., Karen, K., Orly, K., Kinga, H., Kylan, B., Aditi, G., Devin, M., David, S., Jan, B. J., Alice, B. 2024


    In αβ T-cell/CD19 B-cell depleted hematopoietic stem cell transplantation (αβhaplo-HSCT) recipients, anti--thymocyte globulin (ATG, Thymoglobulin®) is used for preventing graft rejection and graft-versus-host disease (GvHD). However, optimal dosing has yet to be established. Here, we present the first comparative analysis of three different ATG dosing strategies and their impact on immune reconstitution and GvHD.Our study aims to evaluate the effects of three distinct dosing strategies of ATG on engraftment success, αβ+ and γδ+ T-cells immune reconstitution, and the incidence and severity of acute GvHD in recipients of αβhaplo-HSCT.This comparative analysis includes three cohorts of pediatric patients with malignant (36) or non-malignant diseases (8). Cohorts 1 and 2 were given fixed ATG doses, while cohort 3 received doses via a new nomogram, based on absolute lymphocyte count (ALC) and body weight (BW).Cohort 3 showed 0% incidence of Day-100 grade II-IV acute GvHD, as opposed to 48% and 27% in cohort 1 and 2, respectively. Further, cohort 3 (the ALC/BW-based cohort) had a significant increase in CD4+ and CD8+ naïve T-cells by Day 90 (P=0.04; P=0.03). Additionally, we found that the reconstitution and maturation of γδ+ T-cells post-HSCT was not impacted across all three cohorts. Cumulative ATG exposure in all cohorts was lower than previously reported in T-cell replete settings, with a lower pre-HSCT exposure (<40 AU*day/mL) correlating with engraftment failure (p=0.007). Conversely, a post-HSCT ATG exposure between 10-15 AU*day/mL was optimal for improving Day-100 CD4+ (p= 0.058) and CD8+ (p=0.03) immune reconstitution, without increasing relapse or non-relapse mortality risks.This study represents the first comparative analysis of ATG Thymoglobulin® exposure in αβhaplo-HSCT recipients. Our findings indicate that i) a 1-2-fold ATG to ATLG bioequivalence is more effective than previously established standards, and ii) ATG exposure post-HSCT does not adversely affect γδ+ T-cell immune reconstitution. Furthermore, a model-based ATG dosing strategy effectively reduces graft rejection and Day-100 acute GvHD while also promoting early CD4+/CD8+ immune reconstitution. These insights suggest that further optimization, including more distal administration of higher ATG doses within an ALC/BW-based strategy, will yield even greater improvements in outcomes.

    View details for DOI 10.1016/j.jtct.2024.05.015

    View details for PubMedID 38768907

  • Extracellular release of mitochondria induced by pre-hematopoietic stem cell transplant conditioning exacerbates GVHD. Blood advances Vijayan, V., Yan, H., Lohmeyer, J. K., Prentiss, K. A., Patil, R. V., Barbarito, G., Lopez, I., Elezaby, A., Peterson, K., Baker, J., Ostberg, N. P., Bertaina, A., Negrin, R. S., Mochly-Rosen, D., Weinberg, K. I., Haileselassie, B. 2024


    Despite therapeutic advancements, GVHD is a major complication of HSCT. In current models of GVHD, tissue injury induced by cytotoxic conditioning regimens, along with translocation of microbes expressing Pathogen Associated Molecular Patterns (PAMPs), result in activation of host antigen-presenting cells (APC) to stimulate alloreactive donor T lymphocytes. Recent studies have demonstrated that in many pathologic states, tissue injury results in the release of mitochondria from the cytoplasm to the extracellular space. We hypothesized that extracellular mitochondria, which are related to archaebacteria, could also trigger GVHD by stimulation of host APC. We found that clinically relevant doses of radiation or busulfan induced extracellular release of mitochondria by various cell types, including cultured intestinal epithelial cells. Conditioning-mediated mitochondrial release was associated with mitochondrial damage and impaired quality control but did not affect the viability of the cells. Extracellular mitochondria directly stimulated host APCs to express higher levels of MHC-II, co-stimulatory CD86, and pro-inflammatory cytokines, resulting in increased donor T cell activation, and proliferation in mixed lymphocyte reactions. Analyses of plasma from both experimental mice and a cohort of children undergoing HSCT demonstrated that conditioning induced extracellular mitochondrial release in vivo. In mice undergoing MHC mismatched HSCT, administration of purified syngeneic extracellular mitochondria increased host APC activation and exacerbated GVHD. Our data suggests that pre-HSCT conditioning results in extracellular release of damaged mitochondria which increase alloreactivity and exacerbate GVHD. Therefore, decreasing the extracellular release of damaged mitochondria following conditioning could serve as a novel strategy for GVHD prevention.

    View details for DOI 10.1182/bloodadvances.2023012328

    View details for PubMedID 38701354

  • Advancements in γδT cell engineering: paving the way for enhanced cancer immunotherapy. Frontiers in immunology Yuan, M., Wang, W., Hawes, I., Han, J., Yao, Z., Bertaina, A. 2024; 15: 1360237


    Comprising only 1-10% of the circulating T cell population, γδT cells play a pivotal role in cancer immunotherapy due to their unique amalgamation of innate and adaptive immune features. These cells can secrete cytokines, including interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α), and can directly eliminate tumor cells through mechanisms like Fas/FasL and antibody-dependent cell-mediated cytotoxicity (ADCC). Unlike conventional αβT cells, γδT cells can target a wide variety of cancer cells independently of major histocompatibility complex (MHC) presentation and function as antigen-presenting cells (APCs). Their ability of recognizing antigens in a non-MHC restricted manner makes them an ideal candidate for allogeneic immunotherapy. Additionally, γδT cells exhibit specific tissue tropism, and rapid responsiveness upon reaching cellular targets, indicating a high level of cellular precision and adaptability. Despite these capabilities, the therapeutic potential of γδT cells has been hindered by some limitations, including their restricted abundance, unsatisfactory expansion, limited persistence, and complex biology and plasticity. To address these issues, gene-engineering strategies like the use of chimeric antigen receptor (CAR) T therapy, T cell receptor (TCR) gene transfer, and the combination with γδT cell engagers are being explored. This review will outline the progress in various engineering strategies, discuss their implications and challenges that lie ahead, and the future directions for engineered γδT cells in both monotherapy and combination immunotherapy.

    View details for DOI 10.3389/fimmu.2024.1360237

    View details for PubMedID 38576617

    View details for PubMedCentralID PMC10991697

  • Vaccine Associated Measles Complicated by Suspected Measles Inclusion Body Encephalitis in a Pediatric Leukemia Patient and Stem Cell Transplant Recipient: A Focus on Clinical Evolution and Management. The Pediatric infectious disease journal Kushner, L. E., Kamens, J., Bertaina, A., Shyr, D., Gans, H. A. 2024


    Immunocompromised individuals are at increased risk for severe disease and complications from viral infections, highlighting the importance of vaccination. However, in extremely rare situations, vaccine associated viral infections can be associated with disseminated disease and complications in immunocompromised hosts.Herein, we present a case of a 1-year-old child diagnosed with acute myeloid leukemia less than 2 weeks after receiving live viral vaccines who developed acute vaccine-strain measles virus disease, later complicated by central nervous system involvement following hematopoietic stem cell transplantation. A brain biopsy specimen was positive for vaccine-strain measles virus detected by reverse transcriptase polymerase chain reaction.She was treated with intravenous ribavirin, inosine pranobex, intrathecal interferon-alpha and donor lymphocyte infusion following measles-mumps-rubella vaccine boost. Despite these measures, the patient suffered neurologic decline and dysautonomia, expiring after compassionate extubation. Management and ideal risk mitigation strategies are discussed within the context of existing literature for this rare complication.

    View details for DOI 10.1097/INF.0000000000004299

    View details for PubMedID 38380931

  • Strategic infection prevention after genetically modified hematopoietic stem cell therapies: recommendations from the International Society for Cell & Gene Therapy Stem Cell Engineering Committee. Cytotherapy John, T. D., Maron, G., Abraham, A., Bertaina, A., Bhoopalan, S. V., Bidgoli, A., Bonfim, C., Coleman, Z., DeZern, A., Li, J., Louis, C., Oved, J., Pavel-Dinu, M., Purtill, D., Ruggeri, A., Russell, A., Wynn, R., Boelens, J. J., Prockop, S., Sharma, A. 2024


    There is lack of guidance for immune monitoring and infection prevention after administration of ex vivo genetically modified hematopoietic stem cell therapies (GMHSCT). We reviewed current infection prevention practices as reported by providers experienced with GMHSCTs across North America and Europe, and assessed potential immunologic compromise associated with the therapeutic process of GMHSCTs described to date. Based on these assessments, and with consensus from members of the International Society for Cell & Gene Therapy (ISCT) Stem Cell Engineering Committee, we propose risk-adapted recommendations for immune monitoring, infection surveillance and prophylaxis, and revaccination after receipt of GMHSCTs. Disease-specific and GMHSCT-specific considerations should guide decision making for each therapy.

    View details for DOI 10.1016/j.jcyt.2024.02.005

    View details for PubMedID 38483362

  • Identification of unstable regulatory and autoreactive effector T cells that are expanded in patients with FOXP3 mutations. Science translational medicine Borna, Š., Lee, E., Nideffer, J., Ramachandran, A., Wang, B., Baker, J., Mavers, M., Lakshmanan, U., Narula, M., Garrett, A. K., Schulze, J., Olek, S., Marois, L., Gernez, Y., Bhatia, M., Chong, H. J., Walter, J., Kitcharoensakkul, M., Lang, A., Cooper, M. A., Bertaina, A., Roncarolo, M. G., Meffre, E., Bacchetta, R. 2023; 15 (727): eadg6822


    Studies of the monogenic autoimmune disease immunodysregulation polyendocrinopathy enteropathy X-linked syndrome (IPEX) have elucidated the essential function of the transcription factor FOXP3 and thymic-derived regulatory T cells (Tregs) in controlling peripheral tolerance. However, the presence and the source of autoreactive T cells in IPEX remain undetermined. Here, we investigated how FOXP3 deficiency affects the T cell receptor (TCR) repertoire and Treg stability in vivo and compared T cell abnormalities in patients with IPEX with those in patients with autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy syndrome (APECED). To study Tregs independently of their phenotype and to analyze T cell autoreactivity, we combined Treg-specific demethylation region analyses, single-cell multiomic profiling, and bulk TCR sequencing. We found that patients with IPEX, unlike patients with APECED, have expanded autoreactive T cells originating from both autoreactive effector T cells (Teffs) and Tregs. In addition, a fraction of the expanded Tregs from patients with IPEX lost their phenotypic and functional markers, including CD25 and FOXP3. Functional experiments with CRISPR-Cas9-mediated FOXP3 knockout Tregs and Tregs from patients with IPEX indicated that the patients' Tregs gain a TH2-skewed Teff-like function, which is consistent with immune dysregulation observed in these patients. Analyses of FOXP3 mutation-carrier mothers and a patient with IPEX after hematopoietic stem cell transplantation indicated that Tregs expressing nonmutated FOXP3 prevent the accumulation of autoreactive Teffs and unstable Tregs. These findings could be directly used for diagnostic and prognostic purposes and for monitoring the effects of immunomodulatory treatments.

    View details for DOI 10.1126/scitranslmed.adg6822

    View details for PubMedID 38117899

  • Stem cell transplantation for ALL: you've always got a donor, why not always use it? Hematology. American Society of Hematology. Education Program Shyr, D., Davis, K. L., Bertaina, A. 2023; 2023 (1): 84-90


    Hematopoietic stem cell transplantation (HSCT) represents a consolidated therapeutic strategy for high-risk pediatric acute lymphoblastic leukemia (ALL), offering the potential for curative treatment. This manuscript delves into the debate around the more universal application of HSCT for pediatric ALL in the modern era, considering the ubiquitous availability of suitable donors. In fact, despite significant advancements in chemotherapy, targeted therapy, and immunotherapy, a subset of pediatric patients with ALL with high-risk features or relapse continue to encounter poor prognostic outcomes. For this subgroup of patients, HSCT often remains the only potentially curative measure, leveraging the graft-versus- leukemia effect for long-term disease control. Nevertheless, the procedure's complexity and associated risks have traditionally curtailed its widespread use. However, the scenario is shifting with improvements in HLA matching, availability of alternative donor sources, less toxic conditioning regimens, and improved supportive care protocols. Concurrently, emerging therapies like CD19+ CAR T cells present new considerations for definitive therapy selection in relapsed/ refractory ALL. This article reviews critical current evidence and debates the potential of HSCT as a more universal treatment for ALL, reevaluating traditional treatment stratification in light of the constant availability of stem cell donors.

    View details for DOI 10.1182/hematology.2023000423

    View details for PubMedID 38066901

  • Engineering the best transplant outcome for high-risk acute myeloid leukemia: the donor, the graft and beyond. Cytotherapy Belbachir, S., Abraham, A., Sharma, A., Prockop, S., DeZern, A. E., Bonfim, C., Bidgoli, A., Li, J., Ruggeri, A., Bertaina, A., Boelens, J. J., Purtill, D. 2023


    Allogeneic hemopoietic cell transplantation remains the goal of therapy for high-risk acute myeloid leukemia (AML). However, treatment failure in the form of leukemia relapse or severe graft-versus-host disease remains a critical area of unmet need. Recently, significant progress has been made in the cell therapy-based interventions both before and after transplant. In this review, the Stem Cell Engineering Committee of the International Society for Cell and Gene Therapy summarizes the literature regarding the identification of high risk in AML, treatment approaches before transplant, optimal transplant platforms and measures that may be taken after transplant to ideally prevent, or, if need be, treat AML relapse. Although some strategies remain in the early phases of clinical investigation, they are built on progress in pre-clinical research and cellular engineering techniques that are already improving outcomes for children and adults with high-risk malignancies.

    View details for DOI 10.1016/j.jcyt.2023.11.004

    View details for PubMedID 38054912

  • Longitudinal Clinical Data Improves Survival Prediction after Hematopoietic Cell Transplantation Using Machine Learning. Blood advances Zhou, Y., Smith, J., Keerthi, D., Li, C., Sun, Y., Mothi, S. S., Shyr, D., Spitzer, B., Harris, A. C., Chatterjee, A., Chatterjee, S., Shouval, R., Naik, S., Bertaina, A., Boelens, J. J., Triplett, B. M., Tang, L., Sharma, A. 2023


    Serial prognostic evaluation of patients after allogeneic hematopoietic cell transplantation (alloHCT) might help identify patients at high risk of developing potentially lethal organ dysfunction. Current prediction algorithms are based on models that do not incorporate changes to the patients' clinical condition that occur after alloHCT in the model development, which limits their predictive ability. We developed and validated a robust risk-prediction algorithm to predict short-term and long-term survival after alloHCT in pediatric patients that includes baseline biological variables, as well as changes in the patients' clinical status after alloHCT. The model was developed using clinical data from children and young adults treated at a single academic quaternary-care referral center. The model was created using a randomly split training dataset (70% of the cohort), internally validated (remaining 30% of the cohort from the same center), and then externally validated on patient data from another tertiary-care referral center. Repeated clinical measurements performed from 30 days before alloHCT to 30 days afterwards were extracted from the electronic medical record and incorporated into the model to predict survival at 100 days, 1-year, and 2-years after alloHCT. Of the 738 patients who underwent their first alloHCT at our institution between 2000 and 2020, 517 (70%) were randomly included in the training dataset and 221 (30%) constituted the validation dataset. When compared with models constructed from baseline variables alone, the naïve-Bayes machine learning models incorporating longitudinal data were significantly better at predicting whether patients would be alive or deceased at the given timepoints. This proof-of-concept study demonstrates that unlike traditional prognostic tools that use fixed variables for risk assessment, incorporating dynamic variability using clinical and laboratory data improves the prediction of mortality in patients undergoing alloHCT.

    View details for DOI 10.1182/bloodadvances.2023011752

    View details for PubMedID 37991991

  • Radiation and Busulfan-Free Hematopoietic Stem Cell Transplantation Using Briquilimab (JSP191) Anti-CD117 Antibody-Conditioning, Transient Immunosuppression and TCR α β + T-Cell/CD19+B-Cell Depleted Haploidentical Grafts in Patients with Fanconi Anemia Agarwal, R., Bertaina, A., Soco, C., Saini, G., Kunte, N., Hiroshima, L., Chan, Y., Willner, H., Krampf, M. L., Nofal, R., Barbarito, G., Sen, S., Felber, M., Van Hentenryck, M., Walck, E., Scheck, A., Thongthip, S., Logan, A. C., Dougall, K., Bouge, A., Boelens, J., Long-Boyle, J. R., Weissman, I. L., Shizuru, J., Pang, W. W., Weinberg, K. I., Parkman, R., Roncarolo, M., Porteus, M., Czechowicz, A. AMER SOC HEMATOLOGY. 2023
  • Targeting of CD25+Activated T Cells in Gvhd-Associated Marrow Suppression and Cytopenias John, T. D., Valine, L., Barbarito, G., Kwon, H., Arai, S., Martin-Kool, B., Kumari, R., Shizuru, J., Oak, J. S., Parkman, R., Bertaina, A., Weinberg, K. I. AMER SOC HEMATOLOGY. 2023
  • Extracellular Release of Damaged Mitochondria Induced By Cytotoxic Conditioning Exacerbates Graft-Versus-Host Disease Vijayan, V., Yan, H., Lohmeyer, J., Prentiss, K., Patil, R., Barbarito, G., Lopez, I., Elezaby, A., Peterson, K., Baker, J., Ostberg, N., Bertaina, A., Negrin, R. S., Mochly-Rosen, D., Weinberg, K. I., Haileselassie, B. AMER SOC HEMATOLOGY. 2023
  • Towards Automated Engineering of Donor-Derived T Lymphocytes into CRISPR/Cas9-Mediated CAR T Cells in a Closed-System Oseghale, A., Bertaina, A., Porteus, M. AMER SOC HEMATOLOGY. 2023
  • T-ALLO10 INFUSION AFTER A.DEPLETED-HSCT IN CHILDREN AND YOUNG ADULTS WITH HEMATOLOGIC MALIGNANCIES: IMPROVED IMMUNE RECONSTITUTION IN THE ABSENCE OF SEVERE GVHD Bertaina, A., Bacchetta, R., Shyr, D., Saini, G., Lee, J., Kristovich, K., Agarwal-Hashmi, R., Klein, O., Melsop, K., Tate, K., Barbarito, G., Oppizzi, L., Chen, P., Cepika, A., Roncarolo, M. SPRINGERNATURE. 2023: 232-234
  • Epigenetic Profiling of PTPN11 Mutant JMML Hematopoietic Stem and Progenitor Cells Reveals an Aberrant Histone Landscape. Cancers Sinha, R., Dvorak, M., Ganesan, A., Kalesinskas, L., Niemeyer, C. M., Flotho, C., Sakamoto, K. M., Lacayo, N., Patil, R. V., Perriman, R., Cepika, A. M., Liu, Y. L., Kuo, A., Utz, P. J., Khatri, P., Bertaina, A. 2023; 15 (21)


    Juvenile myelomonocytic leukemia (JMML) is a deadly pediatric leukemia driven by RAS pathway mutations, of which >35% are gain-of-function in PTPN11. Although DNA hypermethylation portends severe clinical phenotypes, the landscape of histone modifications and chromatin profiles in JMML patient cells have not been explored. Using global mass cytometry, Epigenetic Time of Flight (EpiTOF), we analyzed hematopoietic stem and progenitor cells (HSPCs) from five JMML patients with PTPN11 mutations. These data revealed statistically significant changes in histone methylation, phosphorylation, and acetylation marks that were unique to JMML HSPCs when compared with healthy controls. Consistent with these data, assay for transposase-accessible chromatin with sequencing (ATAC-seq) analysis revealed significant alterations in chromatin profiles at loci encoding post-translational modification enzymes, strongly suggesting their mis-regulated expression. Collectively, this study reveals histone modification pathways as an additional epigenetic abnormality in JMML patient HSPCs, thereby uncovering a new family of potential druggable targets for the treatment of JMML.

    View details for DOI 10.3390/cancers15215204

    View details for PubMedID 37958378

  • Effect of Testicular Boost in Children With Leukemia Receiving Total Body Irradiation and Stem Cell Transplant: A Single-Institution Experience. Advances in radiation oncology Blomain, E. S., Jiang, A., Donaldson, S. S., Agarwal, R., Bertaina, A., Shyr, D., Eisenberg, M. L., Hoppe, R. T., Hiniker, S. M., Oh, J. 2023; 8 (1): 101071


    Children with leukemia who receive fractionated total body irradiation (fTBI) with 12 to 13.2 Gy as part of conditioning for hematopoietic stem cell transplant are frequently treated with an additional 4 Gy testicular boost to reduce the risk of testicular relapse. While institutional practices vary, limited data exists regarding whether the 4-Gy testicular boost reduces the risk of relapse and whether it causes toxicity beyond that imparted by TBI. This study compared the survival and endocrine outcomes among the patients who were treated with and without a testicular boost as part of fTBI from 1990 to 2019 at our center.We retrospectively reviewed charts of male children with leukemia treated with fTBI as part of a conditioning regimen for stem cell transplant from 1990 to 2019. Reported outcomes included progression-free survival, testicular relapse rate, and overall survival. Gonadal dysfunction and fertility were assessed by comparing the rate of abnormally low testosterone or high luteinizing hormone or follicular stimulating hormone, number of offspring, fertility service use, and abnormal sperm count in the subsequent follow-up period between the testicular boost and nonboost subset.Ninety-three male patients (63 acute lymphoblastic leukemia, 30 acute myeloid leukemia) with a median age of 9 years (range, 1-22) and follow-up of 3.3 years were included. In addition to 12- to 13.2-Gy fTBI, 51 male patients (54%) received a testicular boost to 4 Gy. There was 1 testicular relapse in the boost subset and none in the nonboost subset. Five-year progression-free survival for the boost and nonboost subset was 74% and 66%, respectively (P = .31). On multivariable analysis, boost was not associated with improved relapse-free survival or overall survival. More patients in the boost subset (35 of 51, 69%) had abnormal serum gonadal blood work compared with the nonboost subset (18 of 42, 43%) (P = .03).Omission of testicular boost may be associated with comparable oncologic but improved gonadal endocrine outcomes and should be further studied.

    View details for DOI 10.1016/j.adro.2022.101071

    View details for PubMedID 36483061

    View details for PubMedCentralID PMC9723295

  • Immunosuppression and recurrence free parental donor kidney transplant in patient with previous graft loss secondary to recurrent FSGS using prospective a/b T cell, CD19 depleted haploidentical parental stem cell transplant Grimm, P., Wong, C., Goldstein-Fuchs, D., Chaudhuri, A., James, G., Boyd, D., Good, J., Woo, M., Gallo, A., Bertaina, A. SPRINGER. 2023: 2332
  • Precision Delivery of Steroids as a Rescue Therapy for Gastrointestinal Graft-versus-Host Disease in Pediatric Stem Cell Transplant Recipients. Journal of clinical medicine Levitte, S., Ganguly, A., Frolik, S., Guevara-Tique, A. A., Patel, S., Tadas, A., Klein, O., Shyr, D., Agarwal-Hashmi, R., Beach, L., Callard, E., Weinacht, K., Bertaina, A., Thakor, A. S. 2023; 12 (13)


    Graft versus host disease (GVHD) is one of the most serious complications following stem cell transplant in children and is a major cause of morbidity and mortality. Corticosteroids remain the mainstay of treatment, and although a majority of children respond to systemic steroids, those refractory to or dependent upon corticosteroids suffer from complications secondary to long-term steroid administration. This problem has prompted consideration of steroid-sparing treatment strategies, although the time to clinical remission can be variable. Intraarterial corticosteroid delivery has been used in adults as a rescue therapy in steroid-resistant patients, but its use in children has been limited. We investigated the feasibility of intraarterial steroid administration into the bowel and/or liver in a cohort of six pediatric patients with acute GVHD. All patients successfully underwent treatment with no serious adverse effects. Five of five (100%) patients with gastrointestinal bleeding due to GVHD had rapid symptom improvement by 48 h, which was durable up to three weeks. Three of four (75%) patients with hepatic GVHD had improved cholestasis following intraarterial steroid administration. Our experience with this small cohort preliminarily demonstrated the feasibility and safety of intraarterial steroid administration in children with acute GVHD. This approach warrants consideration as a rescue therapy in steroid-refractory cases and as a "bridge" therapy for children with severe acute GVHD who are transitioning to steroid-sparing regimens.

    View details for DOI 10.3390/jcm12134229

    View details for PubMedID 37445274

  • CLONAL ANALYSIS OF.d IMMUNE RECONSTRUCTION POST αβHAPLO-HSCT Barbarito, G., Louis, D. M., Patil, R., Sinha, R., Oppizzi, L., Yao, Z., Weinberg, K., Parkman, R., Bertaina, A. ELSEVIER SCI LTD. 2023: S213-S214
  • T-ALLO10 INFUSION AFTER AßDEPLETED-HSCT IN CHILDREN AND YOUNG ADULTS WITH HEMATOLOGIC MALIGNANCIES: IMPROVED IMMUNE RECONSTITUTION IN THE ABSENCE OF SEVERE GVHD Bertaina, A., Bacchetta, R., Shyr, D. C., Saini, G., Lee, J., Kristovich, K., Agarwal, R., Klein, O., Melsop, K., Tate, K., Barbarito, G., Oppizzi, L., Chen, P., Cepika, A., Roncarolo, M. ELSEVIER SCI LTD. 2023: S123-S124
  • Viral-specific T cells for Cytomegalovirus retinitis following hematopoietic stem cell transplantation: A success story. Pediatric blood & cancer Montiel-Esparza, R., Michalak, S. M., Le, A. H., Or, C., Nguyen, Q. D., Khoury, R., Grimley, M. S., Bertaina, A., Klinger, E., Shah, A. J., Wood, E. H. 2023: e30429


    Cytomegalovirus retinitis (CMVR) following hematopoietic stem cell transplantation (HCT) for a primary immunodeficiency is a rare but highly morbid condition with potential irreversible consequences despite optimal antiviral pharmacotherapy. Viral-specific T cells (VSTs) pose a promising and safe approach eradicating intractable viral disease. We describe the case of a 21-month-old male with Wiskott-Aldrich syndrome (WAS) and CMVR post HCT with sustained long-term virologic and clinical response after CMV-specific T-cell therapy. This case highlights the need to consider VST as an adjunct upfront strategy in refractory CMVR and for routine ophthalmologic screening and surveillance in high-risk patients post HCT.

    View details for DOI 10.1002/pbc.30429

    View details for PubMedID 37243390

  • International society for cell & gene therapy stem cell engineering committee: Cellular therapies for the treatment of graft-versus-host-disease after hematopoietic stem cell transplant. Cytotherapy Garcia-Rosa, M., Abraham, A., Bertaina, A., Bhoopalan, S. V., Bonfim, C., Cohen, S., DeZern, A., Louis, C., Oved, J., Pavel-Dinu, M., Purtill, D., Ruggeri, A., Russell, A., Sharma, A., Wynn, R., Boelens, J. J., Prockop, S. 2023


    Allogeneic hematopoietic stem cell transplant is a curative approach for many malignant and non-malignant hematologic conditions. Despite advances in its prevention and treatment, the morbidity and mortality related to graft-versus-host disease (GVHD) remains. The mechanisms by which currently used pharmacologic agents impair the activation and proliferation of potentially alloreactive T cells reveal pathways essential for the detrimental activities of these cell populations. Importantly, these same pathways can be important in mediating the graft-versus-leukemia effect in recipients transplanted for malignant disease. This knowledge informs potential roles for cellular therapies such as mesenchymal stromal cells and regulatory T cells in preventing or treating GVHD. This article reviews the current state of adoptive cellular therapies focused on GVHD treatment.We conducted a search for scientific literature in PubMed® and ongoing clinical trials in with the keywords "Graft-versus-Host Disease (GVHD)," "Cellular Therapies," "Regulatory T cells (Tregs)," "Mesenchymal Stromal (Stem) Cells (MSCs)," "Natural Killer (NK) Cells," "Myeloid-derived suppressor cells (MDSCs)," and "Regulatory B-Cells (B-regs)." All the published and available clinical studies were included.Although most of the existing clinical data focus on cellular therapies for GVHD prevention, there are observational and interventional clinical studies that explore the potential for cellular therapies to be safe modalities for GVHD treatment while maintaining the graft-versus-leukemia effect in the context of malignant diseases. However, there are multiple challenges that limit the broader use of these approaches in the clinical scenario.There are many ongoing clinical trials to date with the promise to expand our actual knowledge on the role of cellular therapies for GVHD treatment in an attempt to improve GVHD-related outcomes in the near future.

    View details for DOI 10.1016/j.jcyt.2023.02.007

    View details for PubMedID 36941149

  • Transplant for non-malignant disorders: an International Society for Cell & Gene Therapy Stem Cell Engineering Committee report on the role of alternative donors, stem cell sources and graft engineering. Cytotherapy Klein, O. R., Bonfim, C., Abraham, A., Ruggeri, A., Purtill, D., Cohen, S., Wynn, R., Russell, A., Sharma, A., Ciccocioppo, R., Prockop, S., Boelens, J. J., Bertaina, A. 2023


    Hematopoietic stem cell transplantation (HSCT) is curative for many non-malignant disorders. As HSCT and supportive care technologies improve, this life-saving treatment may be offered to more and more patients. With the development of new preparative regimens, expanded alternative donor availability, and graft manipulation techniques, there are many options when choosing the best regimen for patients. Herein the authors review transplant considerations, transplant goals, conditioning regimens, donor choice, and graft manipulation strategies for patients with non-malignant disorders undergoing HSCT.

    View details for DOI 10.1016/j.jcyt.2022.12.005

    View details for PubMedID 36710227

  • Effect of Testicular Boost in Children With Leukemia Receiving Total Body Irradiation and Stem Cell Transplant: A Single-Institution Experience ADVANCES IN RADIATION ONCOLOGY Blomain, E. S., Jiang, A., Donaldson, S. S., Agarwal, R., Bertaina, A., Shyr, D., Eisenberg, M. L., Hoppe, R. T., Hiniker, S. M., Oh, J. 2023; 8 (1)
  • EXTRACELLULAR MITOCHONDRIA EXACERBATE GRAFT-VERSUS-HOST DISEASE MORTALITY IN A MOUSE MODEL Vijayan, V., Yan, H., Lohmeyer, J., Peterson, K., Harden, J., Patil, R., Barbarito, G., Bertaina, A., Negrin, R., Mochly-Rosen, D., Weinberg, K., Haileselassie, B. LIPPINCOTT WILLIAMS & WILKINS. 2023: 640
  • Precision Delivery of Steroids as a Rescue Therapy for Gastrointestinal Graft-versus-Host Disease in Pediatric Stem Cell Transplant Recipients Journal of Clinical Medicine Levitte, s., Ganguly, A., Frolik, S., Guevara-Tique, A., et al 2023; 12 (4229)

    View details for DOI 10.3390/jcm12134229

  • Believe in haploidentical HSCT: less is better. Blood Bertaina, A. 2022; 140 (24): 2523-2524

    View details for DOI 10.1182/blood.2022017535

    View details for PubMedID 36520474

  • Novel Humanized Loss-of-Function NF1 Mouse Model of Juvenile Myelomonocytic Leukemia Sinha, R., Patil, R., Romano, R., Lee, E., Yao, Z., Liu, Y., Porteus, M. H., Bertaina, A. AMER SOC HEMATOLOGY. 2022: 9765-9766
  • Impact of acute GVHD on survival and disease relapse in paediatric patients undergoing allogeneic stem cell transplantation for acute lymphoblastic leukaemia: an EBMT registry-based analysis Lucchini, G., Bertaina, A., Galimard, J. E., Kalwak, K., Dalle, J., Locatelli, F., Taskinen, M., Mirci-Danicar, O., Sedlacek, P., Balduzzi, A., Ghemlas, I., Styczynski, J., Kitra-Roussou, V., Tbakhi, A., Fagioli, F., Bertrand, Y., Aljurf, M., Ifversen, M., Dalissier, A., Bader, P., Corbacioglu, S. SPRINGERNATURE. 2022: 87-88
  • Volumetric Modulated Arc Therapy Total Body Irradiation (VMAT-TBI) in Pediatric and Adolescent/Young Adult Patients Undergoing Stem Cell Transplantation: Early Outcomes and Toxicities Hui, C., Marquez, C., Simiele, E. A., Blomain, E., Oh, J., Bertaina, A., Klein, O., Shyr, D., Jiang, A., Hoppe, R. T., Kovalchuk, N., Hiniker, S. M. ELSEVIER SCIENCE INC. 2022: S123
  • Matching beyond the HLA name: a new classification of HLA-allele mismatches based on peptide binding profile Bertaina, A., Osoegawa, K., Cunanan, K. M., Spellman, S., Fernandez-Vina, M. SPRINGERNATURE. 2022: 94
  • Role of acute GVHD in paediatric patients undergoing allogeneic stem cell transplantation for acute myeloid leukaemia: an imperfect balance Lucchini, G., Bertaina, A., Galimard, J. E., Benakli, M., Kalwak, K., Dalle, J. H., Danicar, O., Ayas, M., Locatelli, F., Skorobogatova, E., Bertrand, Y., Styczynski, J., Tbakhi, A., Sedlacek, P., Balduzzi, A., Ifversen, M., Versluijs, B., Jubert, C., Dalissier, A., Bader, P., Corbacioglu, S. SPRINGERNATURE. 2022: 88
  • Sequential Stem Cell-Kidney Transplantation in Schimke Immuno-osseous Dysplasia Reply NEW ENGLAND JOURNAL OF MEDICINE Bertaina, A., Grimm, P. C., Weinberg, K. 2022; 387 (9): 860
  • Sequential Stem Cell-Kidney Transplantation in Schimke Immuno-osseous Dysplasia. Reply. The New England journal of medicine Bertaina, A., Grimm, P. C., Weinberg, K. 2022; 387 (9): 860

    View details for DOI 10.1056/NEJMc2209527

    View details for PubMedID 36053526

  • Sequential Stem Cell-Kidney Transplantation in Schimke Immuno-osseous Dysplasia. The New England journal of medicine Bertaina, A., Grimm, P. C., Weinberg, K., Parkman, R., Kristovich, K. M., Barbarito, G., Lippner, E., Dhamdhere, G., Ramachandran, V., Spatz, J. M., Fathallah-Shaykh, S., Atkinson, T. P., Al-Uzri, A., Aubert, G., van der Elst, K., Green, S. G., Agarwal, R., Slepicka, P. F., Shah, A. J., Roncarolo, M. G., Gallo, A., Concepcion, W., Lewis, D. B. 2022; 386 (24): 2295-2302


    Lifelong immunosuppression is required for allograft survival after kidney transplantation but may not ultimately prevent allograft loss resulting from chronic rejection. We developed an approach that attempts to abrogate immune rejection and the need for post-transplantation immunosuppression in three patients with Schimke immuno-osseous dysplasia who had both T-cell immunodeficiency and renal failure. Each patient received sequential transplants of alphabeta T-cell-depleted and CD19 B-cell-depleted haploidentical hematopoietic stem cells and a kidney from the same donor. Full donor hematopoietic chimerism and functional ex vivo T-cell tolerance was achieved, and the patients continued to have normal renal function without immunosuppression at 22 to 34 months after kidney transplantation. (Funded by the Kruzn for a Kure Foundation.).

    View details for DOI 10.1056/NEJMoa2117028

    View details for PubMedID 35704481

  • Viral infection in hematopoietic stem cell transplantation: an International Society for Cell & Gene Therapy Stem Cell Engineering Committee review on the role of cellular therapy in prevention and treatment. Cytotherapy Stanojevic, M., Bertaina, A., Bonfim, C., Ciccocioppo, R., Cohen, S., Purtill, D., Ruggieri, A., Russell, A., Sharma, A., Wynn, R., Boelens, J. J., Prockop, S., Abraham, A., ISCT working committee on stem cell engineering 2022


    Despite recent advances in the field of HSCT, viral infections remain a frequent causeof morbidity and mortality among HSCT recipients. Adoptive transfer of viral specific T cells has been successfully used both as prophylaxis and treatment of viral infections in immunocompromised HSCT recipients. Increasingly, precise risk stratification of HSCT recipients with infectious complications should incorporate not only pretransplant clinical criteria, but milestones of immune reconstitution as well. These factors can better identify those at highest risk of morbidity and mortality and identify a population of HSCT recipients in whom adoptive therapy with viral specific T cells should be considered for either prophylaxis or second line treatment early after inadequate response to first line antiviral therapy. Broadening these approaches to improve outcomes for transplant recipients in countries with limited resources is a major challenge. While the principles of risk stratification can be applied, early detection of viral reactivation as well as treatment is challenging in regions where commercial PCR assays and antiviral agents are not readily available.

    View details for DOI 10.1016/j.jcyt.2022.05.010

    View details for PubMedID 35705447

  • Role of Cytokine Secretion Signatures of Donor-derived T Cells and Recipient Serum Cytokine Profiles as Predictive Biomarkers of Acute Graft-Versus-Host Disease in alpha beta T-cell/CD19 B-cell Depleted Hematopoietic Stem Cell Transplant Pediatric Recipients Montiel-Esparza, R., Barbarito, G., Patil, R., Shyr, D., Saini, G., Parkman, R., Liu, Y., Bertaina, A. WILEY. 2022
  • Matching beyond the HLA name: A new classification of HLA-allele mismatches based on peptide binding profile Osoegawa, K., Cunanan, K., Spellman, S., Bertaina, A., Fernandez-Vina, M. A. WILEY. 2022: 436
  • INVESTIGATION APPROACH TO IMPROVE PURITY IN DEPLETED CELLULAR THERAPY PRODUCTS Pham, T., Sukle, N., McLeod, M. G., Beltran, J., Kim, Y., Habib, M., Patel, S., Amiri, Z., Kim, J., Bertaina, A., McIntyre, C. ELSEVIER SCI LTD. 2022: S192
  • BENCH TO BEDSIDE: ENGINEERED AUTOLOGOUS CD4(LVFOXP3) TREG-LIKE CELL PRODUCT FOR PHASE 1 STUDY IN PATIENTS WITH IPEX SYNDROME Nathan, A., Sato, Y., Tate, K., Wani, P., Terry, C., Fazeli, F., Bouge, A., Goodwin, E., Wright, J., Bertaina, A., Klein, O., Agarwal, R., Roncarolo, M., Bacchetta, R. ELSEVIER SCI LTD. 2022: S144
  • Volumetric modulated arc therapy total body irradiation in pediatric and adolescent/young adult patients undergoing stem cell transplantation: Early outcomes and toxicities. Pediatric blood & cancer Marquez, C., Hui, C., Simiele, E., Blomain, E., Oh, J., Bertaina, A., Klein, O., Shyr, D., Jiang, A., Hoppe, R. T., Kovalchuk, N., Hiniker, S. M. 2022: e29689


    INTRODUCTION: Total body irradiation (TBI) is an important component of many conditioning regimens for hematopoietic stem cell transplantation (HSCT), most commonly used in pediatric and adolescent/young adult (AYA) patients. We aimed to evaluate outcomes and toxicities among pediatric and AYA patients treated with TBI utilizing volumetric modulated arc therapy total body irradiation (VMAT-TBI).METHODS: We reviewed pediatric and AYA patients treated with VMAT-TBI at our institution from 2019 to 2021. Data on patient and disease characteristics, treatment details, outcomes and toxicities were collected. Overall survival (OS) and relapse-free survival (RFS) were analyzed using the Kaplan-Meier method.RESULTS: Among 38 patients, 16 (42.1%) were treated with myeloablative regimens and 22 (57.9%) with nonmyeloablative regimens. Median age was 7.2 years (range: 1-27) and median follow-up was 8.7 months (range: 1-21). Lungs Dmean was 7.3 ± 0.3Gy for myeloablative regimens (range: 6.8-7.8). Kidneys were spared to average mean dose of 71.4 ± 4.8% of prescription dose. Gonadal sparing was achieved for patients treated for nonmalignant diseases to Dmean of 0.7 ± 0.1Gy. No patient experienced primary graft failure; one (2.6%) experienced secondary graft failure. The most common grade 1-2 acute toxicities were nausea (68.4%) and fatigue (55.3%). Mucositis was the most common grade 3-4 acute toxicity, affecting 39.5% of patients. There were no cases of pneumonitis or nephrotoxicity attributable to TBI.CONCLUSION: VMAT-TBI offers increased ability to spare organs at risk in pediatric and AYA patients undergoing HSCT, with a favorable acute/subacute toxicity profile and excellent disease control.

    View details for DOI 10.1002/pbc.29689

    View details for PubMedID 35373904

  • Loss of Treg identity in IPEX patients is associated with increase autoreactivity in Teff cells Borna, S., Lee, E., Lakshmanan, U., Narula, M., Schulze, J., Ramachandran, A., Bertaina, A., Meffre, E., Maria, R., Bacchetta, R. SPRINGER/PLENUM PUBLISHERS. 2022: S78-S79
  • An ISCT Stem Cell Engineering Committee Position Statement on Immune Reconstitution: the importance of predictable and modifiable milestones of immune reconstitution to transplant outcomes. Cytotherapy Bertaina, A., Abraham, A., Bonfim, C., Cohen, S., Purtill, D., Ruggeri, A., Weiss, D., Wynn, R., Boelens, J. J., Prockop, S., ISCT Committee on Stem Cell Engineering 2022; 24 (4): 385-392


    Allogeneic stem cell transplantation is a potentially curative therapy for some malignant and non-malignant disease. There have been substantial advances since the approaches first introduced in the 1970s, and the development of approaches to transplant with HLA incompatible or alternative donors has improved access to transplant for those without a fully matched donor. However, success is still limited by morbidity and mortality from toxicity and imperfect disease control. Here we review our emerging understanding of how reconstitution of effective immunity after allogeneic transplant can protect from these events and improve outcomes. We provide perspective on milestones of immune reconstitution that are easily measured and modifiable.

    View details for DOI 10.1016/j.jcyt.2021.09.011

    View details for PubMedID 35331394

  • Sequential hematopoietic stem cell transplantation (HSCT) followed by kidney transplant-3 children who received ABT-cell/CD19 B-cell depleted HSCT followed by kidney transplant from same parental donor are rejection free and immunosuppression free with donor specific tolerance > 1year post kidney transplant Grimm, P., Lewis, D. B., Al-Uzri, A., Fathallah-Shaykh, S., Shah, A., Agarwal, R., Weinberg, K. I., Gallo, A., Concepcion, W., Bertaina, A. WILEY. 2022
  • Curative therapy for hemoglobinopathies: an International Society for Cell & Gene Therapy Stem Cell Engineering Committee review comparing outcomes, accessibility and cost of ex vivo stem cell gene therapy versus allogeneic hematopoietic stem cell transplantation. Cytotherapy Leonard, A., Bertaina, A., Bonfim, C., Cohen, S., Prockop, S., Purtill, D., Russell, A., Boelens, J. J., Wynn, R., Ruggeri, A., Abraham, A. 2021


    Thalassemia and sickle cell disease (SCD) are the most common monogenic diseases in the world and represent a growing global health burden. Management is limited by a paucity of disease-modifying therapies; however, allogeneic hematopoietic stem cell transplantation (HSCT) and autologous HSCT after genetic modification offer patients a curative option. Allogeneic HSCT is limited by donor selection, morbidity and mortality from transplant conditioning, graft-versus-host disease and graft rejection, whereas significant concerns regarding long-term safety, efficacy and cost limit the broad applicability of gene therapy. Here the authors review current outcomes in allogeneic and autologous HSCT for transfusion-dependent thalassemia and SCD and provide our perspective on issues surrounding accessibility and costs as barriers to offering curative therapy to patients with hereditary hemoglobinopathies.

    View details for DOI 10.1016/j.jcyt.2021.09.003

    View details for PubMedID 34879990

  • Aberrant Histone Landscape in Juvenile Myelomonocytic Leukemia Sinha, R., Dvorak, M., Niemeyer, C. M., Sakamoto, K. M., Patil, R., Jutz, P., Liu, Y., Kuo, A., Bertaina, A. AMER SOC HEMATOLOGY. 2021
  • Functional Immune Tolerance Induced By Sequential Hematopoietic Stem Cell-Solid Organ Transplantation Bertaina, A., Barbarito, G., Ramachandran, V. V., Kristovich, K., Lippner, E., Fathallah-Shaykh, S., Al-Uzri, A., Shah, A. J., Aubert, G., Slepicka, P., Oppizzi, L., Agarwal, R., Roncarolo, M., Gallo, A., Concepcion, W., Weinberg, K. I., Parkman, R., Lewis, D. B., Grimm, P. C. AMER SOC HEMATOLOGY. 2021: 1818-+
  • The Determinative Role of Hematopoietic Stem and Progenitor Cell Graft Composition on the Engraftment Kinetics after HSCT Barbarito, G., Dejene, B., Saini, G., Patil, R., Shyr, D. C., Parkman, R., Bertaina, A., Weinberg, K. I. AMER SOC HEMATOLOGY. 2021: 1794-+
  • Combinatorial Cytokine Secretion Signature of Donor-Derived T Cells Infused with the Graft: A New Potential Biomarker of Acute Graft-Versus-Host Disease t-Cell/CD19 B-Cell Depleted Hematopoietic Stem Cell Transplant Recipients Montiel-Esparza, R., Barbarito, G., Peck, S., Bazzano, M., Patil, R., Shyr, D. C., Saini, G., Parkman, R., Liu, Y., Bertaina, A. AMER SOC HEMATOLOGY. 2021
  • Impact of Testicular Boost on Oncologic Outcomes and Late Effects in Pediatric Patients With Leukemia Receiving Fractionated Total Body Irradiation (TBI): A Single-Institution Experience. International journal of radiation oncology, biology, physics Blomain, E., Jiang, A., Donaldson, S. S., Agarwal, R., Bertaina, A., Shyr, D., Shin, D. H., Hoppe, R. T., Hiniker, S. M. 2021; 111 (3S): e177


    PURPOSE/OBJECTIVE(S): Late toxicity of treatment remains a major consideration among survivors of childhood leukemia. Pediatric patients with leukemia who receive fractionated total body irradiation (fTBI) as conditioning prior to stem cell transplantation (SCT) are frequently treated with a 4 Gy testicular boost in addition to fTBI to 12-13.2 Gy to reduce the risk of testicular relapse in this "sanctuary site." However, total body irradiation can be associated with risk of impaired hormonal function; it is unclear if the 4 Gy testicular boost is needed to reduce risk of testicular relapse, and whether it contributes to the toxicity beyond that imparted by fTBI. This study investigated patients both with and without a boost for risk of recurrence, hormonal function, and fertility.MATERIALS/METHODS: We retrospectively reviewed records of boys treated for leukemia with fTBI as part of conditioning for SCT at our institution from 1989-present. We included patients with both AML and ALL in our analysis. We excluded patients who died in the peri-transplant period. Cumulative incidence was tested using Gray's Test for Equality of Cumulative Incidence Functions. Endocrine outcomes were tested using chi square test.RESULTS: We identified 91 boys (age range 9 months - 22 years), 62 with ALL and 29 with AML. Median follow up was 43.4 months. In addition to fTBI to 12-13.2 Gy, 52 patients received a midplane testicular dose of 4 Gy (49 with ALL, 3 with AML), while 39 patients did not receive the testicular boost (13 with ALL, 26 with AML). There was no difference in time to first relapse or cumulative incidence of relapse between the boost and no-boost groups. There were 2 testicular relapses, one ALL patient who received boost and one AML patient who did not receive boost. The risk of abnormal luteinizing hormone (LH) was 65% in the boost group vs 26% in the non-boost group, P = 0.0063. Serum follicle-stimulating hormone, total testosterone, free testosterone, abnormal sperm studies did not differ between groups, nor did the percentage of patients who required endocrine or fertility treatments. We further investigated the subset of patients with ALL and similarly found no difference in risk of relapse between patients who had received a testicular boost and those who had not boost.CONCLUSION: Omission of boost was associated with comparable risk of recurrence and improved endocrine outcomes, specifically LH levels. Our data suggest that omission of testicular boost in the fTBI program is oncologically safe and results in improved hormonal function.

    View details for DOI 10.1016/j.ijrobp.2021.07.667

    View details for PubMedID 34700848

  • Impact of Testicular Boost on Oncologic Outcomes and Late Effects in Pediatric Patients With Leukemia Receiving Fractionated Total Body Irradiation (TBI): A Single-Institution Experience Blomain, E., Jiang, A., Donaldson, S. S., Agarwal, R., Bertaina, A., Shyr, D., Shin, D. H., Hoppe, R. T., Hiniker, S. M. ELSEVIER SCIENCE INC. 2021: E177
  • Alloantigen-specific type 1 regulatory T cells suppress through CTLA-4 and PD-1 pathways and persist long-term in patients. Science translational medicine Chen, P. P., Cepika, A., Agarwal-Hashmi, R., Saini, G., Uyeda, M. J., Louis, D. M., Cieniewicz, B., Narula, M., Amaya Hernandez, L. C., Harre, N., Xu, L., Thomas, B. C., Ji, X., Shiraz, P., Tate, K. M., Margittai, D., Bhatia, N., Meyer, E., Bertaina, A., Davis, M. M., Bacchetta, R., Roncarolo, M. G. 2021; 13 (617): eabf5264


    [Figure: see text].

    View details for DOI 10.1126/scitranslmed.abf5264

    View details for PubMedID 34705520

  • Impact of treosulfan exposure on early and long-term clinical outcome in pediatric allogeneic HSCT recipients: a prospective multicenter study. Transplantation and cellular therapy van der Stoep, M. Y., Bertaina, A., Moes, D. J., Algeri, M., Bredius, R. G., Smiers, F. J., Berghuis, D., Buddingh, E. P., Mohseny, A. B., Guchelaar, H., Locatelli, F., Zwaveling, J., Lankester, A. C. 2021


    BACKGROUND: Treosulfan-based conditioning has gained popularity in pediatric allogeneic hematopoietic stem cell transplantation (HSCT) because of its presumed favorable efficacy and toxicity profile. Treosulfan is used in standardized dosing regimens based on body surface area. The relationship between systemic treosulfan exposure, early and long term clinical outcome in pediatric patients undergoing allogeneic hematopoietic stem cell transplantation (HSCT) for non-malignant diseases is as yet unresolved.OBJECTIVE: In this study we assessed the association between treosulfan exposure and early, and in particular, long term clinical outcomes.STUDY DESIGN: We conducted a multicenter, prospective observational study and included 110 pediatric patients with non-malignant diseases transplanted between 2011 and 2019 in Leiden, The Netherlands and Rome, Italy. Blood samples were collected and treosulfan area under the curve (AUC0-) was estimated as a measure of exposure. Cox proportional hazard survival analyses were performed to assess the relationship between treosulfan exposure, OS and EFS. The predictive value of systemic treosulfan exposure for the occurrence of toxicity within 28 days is evaluated using a multivariable logistic regression analysis.RESULTS: In the overall cohort, overall survival (OS) and event-free survival (EFS) at 2 years were 89.0% and 75.3%, respectively, with an excellent OS of 97% in children under the age of 2 years. The occurrence of grade II-IV aGvHD, the level of 1-year whole blood chimerism, and 2-year OS and EFS were not correlated with treosulfan exposure. The occurrence of skin toxicity (odds ratio (OR) 3.97, 95% confidence interval (CI) 1.26-13.68, p=0.02) and all grade mucositis (OR 4.43, 95%CI 1.43-15.50, p=0.02), but not ≥ grade 2 mucositis (OR 1.51, 95%CI 0.52-4.58, p=0.46) was related to high treosulfan exposure (>1750 mg*h/L).CONCLUSION: Our study demonstrates that standardized treosulfan-based conditioning results in a favorable OS and EFS in infants and children with non-malignant diseases, independent of interindividual variation in treosulfan exposure. These outcomes can be achieved without the need for therapeutic drug monitoring (TDM), thereby emphasizing the advantage of treosulfan use in this category of patients. Although higher treosulfan exposure increases the risk of skin toxicity, there is no absolute necessity for therapeutic drug monitoring if proper preventive skin measures are taken. More research is needed to assess whether de-escalation of treosulfan doses is possible in order to minimize early and long term toxicity without compromising efficacy.

    View details for DOI 10.1016/j.jtct.2021.09.018

    View details for PubMedID 34607071

  • Outcomes of pediatric patients with therapy-related myeloid neoplasms. Bone marrow transplantation Sharma, A., Huang, S., Li, Y., Brooke, R. J., Ahmed, I., Allewelt, H. B., Amrolia, P., Bertaina, A., Bhatt, N. S., Bierings, M. B., Bies, J., Brisset, C., Brondon, J. E., Dahlberg, A., Dalle, J., Eissa, H., Fahd, M., Gassas, A., Gloude, N. J., Goebel, W. S., Goeckerman, E. S., Harris, K., Ho, R., Hudspeth, M. P., Huo, J. S., Jacobsohn, D., Kasow, K. A., Katsanis, E., Kaviany, S., Keating, A. K., Kernan, N. A., Ktena, Y. P., Lauhan, C. R., Lopez-Hernandez, G., Martin, P. L., Myers, K. C., Naik, S., Olaya-Vargas, A., Onishi, T., Radhi, M., Ramachandran, S., Ramos, K., Rangarajan, H. G., Roehrs, P. A., Sampson, M. E., Shaw, P. J., Skiles, J. L., Somers, K., Symons, H. J., de Tersant, M., Uber, A. N., Versluys, B., Cheng, C., Triplett, B. M. 2021


    Long-term outcomes after allogeneic hematopoietic cell transplantation (HCT) for therapy-related myeloid neoplasms (tMNs) are dismal. There are few multicenter studies defining prognostic factors in pediatric patients with tMNs. We have accumulated the largest cohort of pediatric patients who have undergone HCT for a tMN to perform a multivariate analysis defining factors predictive of long-term survival. Sixty-eight percent of the 401 patients underwent HCT using a myeloablative conditioning (MAC) regimen, but there were no statistically significant differences in the overall survival (OS), event-free survival (EFS), or cumulative incidence of relapse and non-relapse mortality based on the conditioning intensity. Among the recipients of MAC regimens, 38.4% of deaths were from treatment-related causes, especially acute graft versus host disease (GVHD) and end-organ failure, as compared to only 20.9% of deaths in the reduced-intensity conditioning (RIC) cohort. Exposure to total body irradiation (TBI) during conditioning and experiencing grade III/IV acute GVHD was associated with worse OS. In addition, a diagnosis of therapy-related myelodysplastic syndrome and having a structurally complex karyotype at tMN diagnosis were associated with worse EFS. Reduced-toxicity (but not reduced-intensity) regimens might help to decrease relapse while limiting mortality associated with TBI-based HCT conditioning in pediatric patients with tMNs.

    View details for DOI 10.1038/s41409-021-01448-x

    View details for PubMedID 34480120

  • Correction: Myeloablative conditioning for allo-HSCT in pediatric ALL: FTBI or chemotherapy?-A multicenter EBMT-PDWP study. Bone marrow transplantation Willasch, A. M., Peters, C., Sedlacek, P., Dalle, J., Kitra-Roussou, V., Yesilipek, A., Wachowiak, J., Lankester, A., Prete, A., Hamidieh, A. A., Ifversen, M., Buechner, J., Krivan, G., Hamladji, R., Diaz-de-Heredia, C., Skorobogatova, E., Michel, G., Locatelli, F., Bertaina, A., Veys, P., Dupont, S., Or, R., Gungor, T., Aleinikova, O., Sufliarska, S., Sundin, M., Rascon, J., Kaare, A., Nemet, D., Fagioli, F., Klingebiel, T. E., Styczynski, J., Bierings, M., Nagy, K., Abecasis, M., Afanasyev, B., Ansari, M., Vettenranta, K., Alseraihy, A., Chybicka, A., Robinson, S., Bertrand, Y., Kupesiz, A., Ghavamzadeh, A., Campos, A., Pichler, H., Dalissier, A., Labopin, M., Corbacioglu, S., Balduzzi, A., Galimard, J., Bader, P., EBMT Paediatric Diseases Working Party 2021

    View details for DOI 10.1038/s41409-021-01424-5

    View details for PubMedID 34413474

  • Standardizing Definitions of Hematopoietic Recovery, Graft Rejection, Graft Failure, Poor Graft Function, and Donor Chimerism in Allogeneic Hematopoietic Cell Transplantation: A Report on Behalf of the American Society for Transplantation and Cellular Therapy. Transplantation and cellular therapy Kharfan-Dabaja, M. A., Kumar, A., Ayala, E., Aljurf, M., Nishihori, T., Marsh, R., Burroughs, L. M., Majhail, N., Al-Homsi, A. S., Al-Kadhimi, Z. S., Bar, M., Bertaina, A., Boelens, J. J., Champlin, R., Chaudhury, S., DeFilipp, Z., Dholaria, B., El-Jawahri, A., Fanning, S., Fraint, E., Gergis, U., Giralt, S., Hamilton, B. K., Hashmi, S. K., Horn, B., Inamoto, Y., Jacobsohn, D. A., Jain, T., Johnston, L., Kanate, A. S., Kansagra, A., Kassim, A., Kean, L. S., Kitko, C. L., Knight-Perry, J., Kurtzberg, J., Liu, H., MacMillan, M. L., Mahmoudjafari, Z., Mielcarek, M., Mohty, M., Nagler, A., Nemecek, E., Olson, T. S., Oran, B., Perales, M., Prockop, S. E., Pulsipher, M. A., Pusic, I., Riches, M. L., Rodriguez, C., Romee, R., Rondon, G., Saad, A., Shah, N., Shaw, P. J., Shenoy, S., Sierra, J., Talano, J., Verneris, M. R., Veys, P., Wagner, J. E., Savani, B. N., Hamadani, M., Carpenter, P. A. 2021; 27 (8): 642-649


    Allogeneic hematopoietic cell transplantation (allo-HCT) is potentially curative for certain hematologic malignancies and nonmalignant diseases. The field of allo-HCT has witnessed significant advances, including broadening indications for transplantation, availability of alternative donor sources, less toxic preparative regimens, new cell manipulation techniques, and novel GVHD prevention methods, all of which have expanded the applicability of the procedure. These advances have led to clinical practice conundrums when applying traditional definitions of hematopoietic recovery, graft rejection, graft failure, poor graft function, and donor chimerism, because these may vary based on donor type, cell source, cell dose, primary disease, graft-versus-host disease (GVHD) prophylaxis, and conditioning intensity, among other variables. To address these contemporary challenges, we surveyed a panel of allo-HCT experts in an attempt to standardize these definitions. We analyzed survey responses from adult and pediatric transplantation physicians separately. Consensus was achieved for definitions of neutrophil and platelet recovery, graft rejection, graft failure, poor graft function, and donor chimerism, but not for delayed engraftment. Here we highlight the complexities associated with the management of mixed donor chimerism in malignant and nonmalignant hematologic diseases, which remains an area for future research. We recognize that there are multiple other specific, and at times complex, clinical scenarios for which clinical management must be individualized.

    View details for DOI 10.1016/j.jtct.2021.04.007

    View details for PubMedID 34304802

  • SEQUENTIAL HEMATOPOIETIC STEM CELL AND KIDNEY TRANSPLANTATION IN SCHIMKE IMMUNO-OSSEOUS DYSPLASIA: TOWARDS A MODEL FOR ESTABLISHING FUNCTIONAL IMMUNE TOLERANCE FOR SOLID ORGAN TRANSPLANTATION Bertaina, A., Grimm, P., Kristovich, K., Barbarito, G., Lippner, E., Fathallah-Shaykh, S., Al-Uzri, A., van der Elst, K., Agarwal, R., Selpicka, P., Shah, A., Weinberg, K., Parkman, R., Roncarolo, M., Gallo, A., Conception, W., Lewis, D. ELSEVIER SCI LTD. 2021: S136
  • Adoptively Transferred, In Vitro-Generated Alloantigen-Specific Type 1 Regulatory T (Tr1) Cells Persist Long-Term In Vivo Cepika, A., Chen, P. P., Agarwal, R., Saini, G., Louis, D. M., Amaya-Hernandez, L. C., Xu, L., Shiraz, P., Tate, K. M., Margittai, D., Bhatia, N., Meyer, E., Bertaina, A., Davis, M. M., Bacchetta, R., Roncarolo, M. CELL PRESS. 2021: 73
  • Preclinical Safety and Efficacy Validation of CD4(LVFOXP3) Cells as an Innovative Cell-Based Gene Therapy Approach for IPEX Syndrome Sato, Y., Nathan, A., Wright, J., Tate, K., Wani, P., Fazeli, F., Timnak, A., Bhatia, N., Agarwal-Hashmi, R., Bertaina, A., Roncarolo, M., Bacchetta, R. CELL PRESS. 2021: 340
  • Toxoplasmosis in Pediatric Hematopoietic Stem Cell Transplantation Patients. Transplantation and cellular therapy Schwenk, H. T., Khan, A., Kohlman, K., Bertaina, A., Cho, S., Montoya, J. G., Contopoulos-Ioannidis, D. G. 2021; 27 (4): 292–300


    Infection due to the protozoa Toxoplasma gondii can be life-threatening in hematopoietic stem cell transplantation (HSCT) recipients. Most cases of toxoplasmosis in HSCT recipients result from reactivation of latent infection in individuals who were Toxoplasma-seropositive before transplantation and did not receive appropriate prophylaxis. Pretransplantation screening with Toxoplasma IgG and IgM antibodies is suggested for all allogeneic HSCT recipients and their donors and all autologous HSCT recipients. Prevention of toxoplasmosis in T. gondii-seropositive HSCT recipients requires primary prophylaxis, preemptive screening, or both. Trimethoprim-sulfamethoxazole (TMP-SMX) is the preferred agent for Toxoplasma prophylaxis and should be continued for 6 months or until the patient is no longer receiving immunosuppression, whichever is longer, assuming that immune reconstitution has occurred. Preemptive weekly screening with whole blood Toxoplasma PCR should be considered for seropositive HSCT recipients if prophylaxis cannot be given or if prophylaxis other than TMP-SMX is used. The signs, symptoms, and radiographic findings of toxoplasmosis in HSCT recipients can be nonspecific, and the diagnosis requires a high degree of suspicion. Common presentations include fever, encephalopathy with mental status changes or seizures, and pneumonia. A Toxoplasma PCR analysis from whole blood (and other body fluids/tissues according to clinical symptoms) should be obtained in patients in whom there is a concern for toxoplasmosis. Treatment with oral pyrimethamine, sulfadiazine, and leucovorin for at least 6 weeks is the first-line therapy and should be followed by secondary prophylaxis. In this article, we review the published literature regarding the epidemiology, clinical presentation, treatment, and prevention of toxoplasmosis in HSCT recipients.

    View details for DOI 10.1016/j.jtct.2020.11.003

    View details for PubMedID 33840441

  • Response to Trimethoprim-Sulfamethoxazole in a Pediatric Hematopoietic Stem Cell Transplant Recipient With Disseminated Toxoplasmosis: A Case Report. Journal of the Pediatric Infectious Diseases Society Khan, A., Schwenk, H. T., Kohlman, K., Bertaina, A., Cho, S., Montoya, J. G., Contopoulos-Ioannidis, D. 2021


    We describe the presentation and treatment of a patient who developed ongoing fever and diagnosed with disseminated toxoplasmosis post-hematopoietic stem cell transplantation. He was initially treated with trimethoprim-sulfamethoxazole (TMP-SMX) and there was dramatic improvement in his fever curve. He successfully completed a modified course of therapy.

    View details for DOI 10.1093/jpids/piab006

    View details for PubMedID 33693793

  • Toxoplasmosis Among 38,751 Hematopoietic Stem Cell Transplant Recipients: A Systematic Review of Disease Prevalence and a Compilation of Imaging and Autopsy Findings. Transplantation Contopoulos-Ioannidis, D. G., Cho, S. M., Bertaina, A., Leung, A. N., Fischbein, N., Lanzman, B., Schwenk, H. T., Montoya, J. G. 2021


    BACKGROUND: Toxoplasmosis in hematopoietic stem cell transplant-recipients (HSCT) can be life threatening if not promptly diagnosed and treated.METHODS: We performed a systematic review (PubMed last search 03/29/2020) of toxoplasmosis among HSCT-recipients and calculated the toxoplasmosis prevalence across studies. We also created a compilation list of brain imaging, chest imaging and autopsy findings of toxoplasmosis among HSCT-recipients.RESULTS: We identified 46 eligible studies (47 datasets) with 399 toxoplasmosis cases among 38751 HSCT-recipients. There was large heterogeneity in the reported toxoplasmosis prevalence across studies, thus formal meta-analysis was not attempted. The median toxoplasmosis prevalence among 38751 HSCT-recipients was 2.14% (range 0-66.67%). Data on toxoplasmosis among at-risk R+HSCT-recipients were more limited (25 studies; 2404 R+HSCT-recipients [6.2% of all HSCT-recipients]) although the median number of R+HSCT-recipients was 56.79% across all HSCT-recipients. Median toxoplasmosis prevalence across studies among 2404 R+HSCT was 7.51% (range 0-80%) vs 0% (range 0-1.23%) among 7438 R-HSCT. There were limited data to allow meaningful analyses of toxoplasmosis prevalence according to prophylaxis-status of R+HSCT-recipients.CONCLUSION: Toxoplasmosis prevalence among HSCT-recipients is underestimated. The majority of studies report toxoplasmosis prevalence among all HSCT-recipients rather than only among the at-risk R+HSCT-recipients. In fact, the median toxoplasmosis prevalence among all R+/R- HSCT-recipients is 3.5-fold lower compared to the prevalence among only the at-risk R+HSCT-recipients and the median prevalence among R+HSCT-recipients is 7.51-fold higher than among R-HSCT-recipients. The imaging findings of toxoplasmosis among HSCT-recipients can be atypical. High-index of suspicion is needed in R+HSCT-recipients with fever, pneumonia or encephalitis.

    View details for DOI 10.1097/TP.0000000000003662

    View details for PubMedID 33654004

  • TCRαβ/CD19 depleted HSCT from an HLA-haploidentical relative to treat children with different non-malignant disorders. Blood advances Merli, P., Pagliara, D., Galaverna, F., Li Pira, G., Andreani, M., Leone, G., Amodio, D., Pinto, R. M., Bertaina, A., Bertaina, V., Mastronuzzi, A., Strocchio, L., Boccieri, E., Pende, D., Falco, M., Di Nardo, M., Del Bufalo, F., Algeri, M., Locatelli, F. 2021


    Several non-malignant disorders (NMDs), either inherited or acquired, can be cured by allogeneic hematopoietic stem cell transplantation (HSCT). Between January 2012 and April 2020, 70 consecutive children affected by primary immunodeficiencies, inherited/acquired bone marrow failure syndromes, red blood cell disorders or metabolic diseases, lacking a fully-matched donor or requiring urgent transplantation, underwent TCRαβ/CD19-depleted haploidentical HSCT from an HLA-partially matched relative as part of a prospective study (#NCT01810120). Median age at transplant was 3.5 years (range 0.3-16.1); median time from diagnosis to transplant was 10.5 months (2.7 for SCID patients). Primary engraftment was obtained in 51 patients, while 19 and 2 patients experienced either primary or secondary graft failure (GF), the overall incidence of this complication being 30.4%. Most GFs were observed in children with disease at risk for this complication (e.g., aplastic anemia, thalassemia). All but 5 patients experiencing GF were successfully retransplanted. Six patients died of infectious complications (4 had active/recent infections at time of HSCT), the cumulative incidence of transplant-related mortality (TRM) being 8.5%. Cumulative incidence of grade I-II acute GvHD was 14.4% (no patient developed grade III-IV acute GVHD). Only one patient at risk developed mild chronic GvHD. With a median follow-up of 3.5 years, the 5-year probability of overall and disease-free survival was 91.4% and 86.8%, respectively. In conclusion, TCRαβ/CD19-depleted haploidentical HSCT from an HLA-partially matched relative is confirmed to be an effective treatment for children with NMDs. Prompt donor availability, low incidence of GvHD and TRM make this strategy an attractive option in NMDs patients.

    View details for DOI 10.1182/bloodadvances.2021005628

    View details for PubMedID 34592755

  • Harnessing Mechanisms of Immune Tolerance to Improve Outcomes in Solid Organ Transplantation: A Review. Frontiers in immunology Slepicka, P. F., Yazdanifar, M., Bertaina, A. 2021; 12: 688460


    Survival after solid organ transplantation (SOT) is limited by chronic rejection as well as the need for lifelong immunosuppression and its associated toxicities. Several preclinical and clinical studies have tested methods designed to induce transplantation tolerance without lifelong immune suppression. The limited success of these strategies has led to the development of clinical protocols that combine SOT with other approaches, such as allogeneic hematopoietic stem cell transplantation (HSCT). HSCT prior to SOT facilitates engraftment of donor cells that can drive immune tolerance. Recent innovations in graft manipulation strategies and post-HSCT immune therapy provide further advances in promoting tolerance and improving clinical outcomes. In this review, we discuss conventional and unconventional immunological mechanisms underlying the development of immune tolerance in SOT recipients and how they can inform clinical advances. Specifically, we review the most recent mechanistic studies elucidating which immune regulatory cells dampen cytotoxic immune reactivity while fostering a tolerogenic environment. We further discuss how this understanding of regulatory cells can shape graft engineering and other therapeutic strategies to improve long-term outcomes for patients receiving HSCT and SOT.

    View details for DOI 10.3389/fimmu.2021.688460

    View details for PubMedID 34177941

  • Supportive Care During Pediatric Hematopoietic Stem Cell Transplantation: Prevention of Infections. A Report From Workshops on Supportive Care of the Paediatric Diseases Working Party (PDWP) of the European Society for Blood and Marrow Transplantation (EBMT). Frontiers in pediatrics Ifversen, M., Meisel, R., Sedlacek, P., Kalwak, K., Sisinni, L., Hutt, D., Lehrnbecher, T., Balduzzi, A., Diesch, T., Jarisch, A., Gungor, T., Stein, J., Yaniv, I., Bonig, H., Kuhlen, M., Ansari, M., Nava, T., Dalle, J., Diaz-de-Heredia, C., Trigoso, E., Falkenberg, U., Hartmann, M., Deiana, M., Canesi, M., Broggi, C., Bertaina, A., Gibson, B., Krivan, G., Vettenranta, K., Matic, T., Buechner, J., Lawitschka, A., Peters, C., Yesilipek, A., Yalcin, K., Lucchini, G., Bakhtiar, S., Turkiewicz, D., Niinimaki, R., Wachowiak, J., Cesaro, S., Dalissier, A., Corbacioglu, S., Willasch, A. M., Bader, P. 2021; 9: 705179


    Specific protocols define eligibility, conditioning, donor selection, graft composition and prophylaxis of graft vs. host disease for children and young adults undergoing hematopoietic stem cell transplant (HSCT). However, international protocols rarely, if ever, detail supportive care, including pharmaceutical infection prophylaxis, physical protection with face masks and cohort isolation or food restrictions. Supportive care suffers from a lack of scientific evidence and implementation of practices in the transplant centers brings extensive restrictions to the child's and family's daily life after HSCT. Therefore, the Board of the Pediatric Diseases Working Party (PDWP) of the European Society for Blood and Marrow Transplantation (EBMT) held a series of dedicated workshops since 2017 with the aim of initiating the production of a set of minimal recommendations. The present paper describes the consensus reached within the field of infection prophylaxis.

    View details for DOI 10.3389/fped.2021.705179

    View details for PubMedID 34395344

  • The Role of Donor KIR Genotyping in Pediatric Hematopoietic Stem Cell Transplantation For Non-Malignant Disorders Mariano, L., Bertaina, A., Zhang, B. M., Kristovich, K., Vina, M. SPRINGERNATURE. 2020: 640
  • Anti-T-Lymphocyte Globulin (ATLG) and Rituximab for Immunomodulation of Graft-Versus-Host Disease and Graft Failure in Patients with Non-Malignant Disorders: Results of a Multicentre, Randomized, Open-Label Study Algeri, M., Galimberti, S., Bernardo, M., Rovelli, A., Zecca, M., La Nasa, G., Marktel, S., Merli, P., Bertaina, A., Pagliara, D., Boccieri, E., Del Bufalo, F., Gaspari, S., Ruggeri, A., Capitoli, G., Valsecchi, M., Locatelli, F. SPRINGERNATURE. 2020: 85–86
  • HLA-haplotype loss after TCRalphabeta/CD19-depleted haploidentical HSCT. Bone marrow transplantation Shyr, D. C., Zhang, B. M., Saini, G., Madani, N. D., Schultz, L. M., Patel, S., Kristovich, K., Fernandez-Vina, M., Bertaina, A. 2020

    View details for DOI 10.1038/s41409-020-01081-0

    View details for PubMedID 33070150

  • Identification of dual positive CD19+/CD3+ T cells in a leukapheresis product undergoing CAR transduction: a case report. Journal for immunotherapy of cancer Schultz, L., Patel, S., Davis, K. L., Ramakrishna, S., Sahaf, B., Bhatia, N., Baggott, C., Erickson, C., Majzner, R. G., Oak, J., Bertaina, A., Mackall, C., Feldman, S. 2020; 8 (2)


    BACKGROUND: Chimeric antigen receptor (CAR) therapy and hematopoietic stem cell transplantation (HSCT) are therapeutics for relapsed acute lymphocytic leukemia (ALL) that are increasingly being used in tandem. We identified a non-physiologic CD19+/CD3+ T-cell population in the leukapheresis product of a patient undergoing CAR T-cell manufacturing who previously received a haploidentical HSCT, followed by infusion of a genetically engineered T-cell addback product. We confirm and report the origin of these CD19+/CD3+ T cells that have not previously been described in context of CAR T-cell manufacturing. We additionally interrogate the fate of these CD19-expressing cells as they undergo transduction to express CD19-specific CARs.MAIN BODY: We describe the case of a preteen male with multiply relapsed B-ALL who was treated with sequential cellular therapies. He received an alphabeta T-cell depleted haploidentical HSCT followed by addback of donor-derived T cells genetically modified with a suicide gene for iCaspase9 and truncated CD19 for cell tracking (RivoCel). He relapsed 6months following HSCT and underwent leukapheresis and CAR T-cell manufacturing. During manufacturing, we identified an aberrant T-cell population dually expressing CD19 and CD3. We hypothesized that these cells were RivoCel cells and confirmed using flow cytometry and PCR that the identified cells were in fact RivoCel cells and were eliminated with iCaspase9 activation. We additionally tracked these cells through CD19-specific CAR transduction and notably did not detect T cells dually positive for CD19 and CD19-directed CARs. The most likely rationale for this is in vitro fratricide of the CD19+ 'artificial' T-cell population by the CD19-specific CAR+ T cells in culture.CONCLUSIONS: We report the identification of CD19+/CD3+ cells in an apheresis product undergoing CAR transduction derived from a patient previously treated with a haploidentical transplant followed by RivoCel addback. We aim to bring attention to this cell phenotype that may be recognized with greater frequency as CAR therapy and engineered alphabetahaplo-HSCT are increasingly coupled. We additionally suggest consideration towards using alternative markers to CD19 as a synthetic identifier for post-transplant addback products, as CD19-expression on effector T cells may complicate subsequent treatment using CD19-directed therapy.

    View details for DOI 10.1136/jitc-2020-001073

    View details for PubMedID 32929049

  • Sustained fetal hematopoiesis causes juvenile death from leukemia: evidence from a dual-age-specific mouse model. Blood advances Vara, N., Liu, Y., Yan, Y., Lensing, S. Y., Colorado, N., Robinson, D., Zhang, J., Zhang, X., Peterson, E. A., Baltz, N. J., Zhou, D., Bertaina, A., Johann, D. J., Emanuel, P. D., Liu, Y. L. 2020; 4 (15): 3728–40


    It is not clear whether disrupted age-specific hematopoiesis contributes to the complex manifestations in leukemia patients who carry identical mutations, particularly in pediatric and adult patients with similar clinical characteristics. By studying a dual-age-specific mouse model, we demonstrate that (1) loss of Pten during the fetal-to-adult hematopoiesis switch (hematopoiesis switch) causes sustained fetal hematopoiesis, resulting in death in juvenile leukemia; (2) myeloid-biased hematopoiesis in juvenile mice is associated with the sustained fetal properties of hematopoietic stem cells (HSCs); (3) the age specificity of juvenile myelomonocytic leukemia depends on the copy number of Pten and Nf1; (4) single-allelic Pten deletion during the hematopoiesis switch causes constitutive activation of MAPK in juvenile mice with Nf1 loss of heterozygosity (LOH); and (5) Nf1 LOH causes monocytosis in juvenile mice with Pten haploinsufficiency but does not cause lethality until adulthood. Our data suggest that 1 copy of Pten is sufficient to maintain an intact negative-feedback loop of the Akt pathway and HSC function in reconstitution, despite MAPK being constitutively activated in juvenile Pten+/DeltaNf1LOH mice. However, 2 copies of Pten are required to maintain the integrity of the MAPK pathway in juvenile mice with Nf1 haploinsufficiency. Our data indicate that previous investigations of Pten function in wild-type mice may not reflect the impact of Pten loss in mice with Nf1 mutations or other genetic defects. We provide a proof of concept that disassociated age-specific hematopoiesis contributes to leukemogenesis and pediatric demise.

    View details for DOI 10.1182/bloodadvances.2020002326

    View details for PubMedID 32777070

  • A study assessing the feasibility of randomization of pediatric and young adult patients between matched unrelated donor bone marrow transplantation and immune-suppressive therapy for newly diagnosed severe aplastic anemia: A joint pilot trial of the North American Pediatric Aplastic Anemia Consortium and the Pediatric Transplantation and Cellular Therapy Consortium. Pediatric blood & cancer Pulsipher, M. A., Lehmann, L. E., Bertuch, A. A., Sasa, G., Olson, T., Nakano, T., Gilio, A., Burroughs, L. M., Lipton, J. M., Huang, J. N., Dickerson, K., Bertaina, A., Zhuang, C., Malsch, M., Fleming, M., Weller, E., Shimamura, A., Williams, D. A. 2020: e28444


    BACKGROUND: Recent data show survival after matched unrelated donor (MUD) bone marrow transplantation (BMT) is similar to matched sibling procedures for young patients with severe aplastic anemia (SAA). Donor delays, risk of transplant-related mortality (TRM), and concern about chronic graft versus host disease raise questions about whether MUD BMT or immune suppression therapy (IST) should be preferred initial therapy for young patients lacking matched sibling donors.PROCEDURE: We performed a pilot trial to assess the feasibility of randomizing patients under age 26 with newly diagnosed SAA to receive IST versus MUD BMT. Primary aims assessed the acceptability of randomization and timing of BMT. Secondary aims measured toxicities, response, and survival.RESULTS: Sixty-seven patients with possible SAA were screened at nine centers. Of 57 with confirmed SAA, 23 underwent randomization and received therapy with a median follow-up of 18 months. Of 12 randomized to BMT, 10 started BMT as initial therapy at a median of 36 days after randomization. One BMT recipient experienced secondary graft failure, requiring a second procedure. Six of 11 randomized to IST responded, whereas five with refractory disease underwent successful salvage BMT. One patient achieving complete response relapsed after discontinuation of immune suppression and died of infection after salvage BMT.CONCLUSIONS: This feasibility study showed that a high percentage of patients underwent randomization and received up-front MUD BMT. Our study lays the groundwork for a larger randomized trial that will define best initial therapy for young patients with SAA who have an available MUD.

    View details for DOI 10.1002/pbc.28444

    View details for PubMedID 32776425

  • Phenotypic and Functional Characterization of NK Cells in alpha beta T-Cell and B-Cell Depleted Haplo-HSCT to Cure Pediatric Patients with Acute Leukemia CANCERS Meazza, R., Falco, M., Loiacono, F., Canevali, P., Della Chiesa, M., Bertaina, A., Pagliara, D., Merli, P., Indio, V., Galaverna, F., Algeri, M., Moretta, F., Colomar-Carando, N., Muccio, L., Sivori, S., Pession, A., Mingari, M., Moretta, L., Moretta, A., Locatelli, F., Pende, D. 2020; 12 (8)


    NK cells can exert remarkable graft-versus-leukemia (GvL) effect in HLA-haploidentical hematopoietic stem cell transplantation (haplo-HSCT). Here, we dissected the NK-cell repertoire of 80 pediatric acute leukemia patients previously reported to have an excellent clinical outcome after αβT/B-depleted haplo-HSCT. This graft manipulation strategy allows the co-infusion of mature immune cells, mainly NK and γδT cells, and hematopoietic stem cells (HSCs). To promote NK-cell based antileukemia activity, 36/80 patients were transplanted with an NK alloreactive donor, defined according to the KIR/KIR-Ligand mismatch in the graft-versus-host direction. The analysis of the reconstituted NK-cell repertoire in these patients showed relatively high proportions of mature and functional KIR+NKG2A-CD57+ NK cells, including the alloreactive NK cell subset, one month after HSCT. Thus, the NK cells adoptively transfused with the graft persist as a mature source of effector cells while new NK cells differentiate from the donor HSCs. Notably, the alloreactive NK cell subset was endowed with the highest anti-leukemia activity and its size in the reconstituted repertoire could be influenced by human cytomegalovirus (HCMV) reactivation. While the phenotypic pattern of donor NK cells did not impact on post-transplant HCMV reactivation, in the recipients, HCMV infection/reactivation fostered a more differentiated NK-cell phenotype. In this cohort, no significant correlation between differentiated NK cells and relapse-free survival was observed.

    View details for DOI 10.3390/cancers12082187

    View details for Web of Science ID 000578885400001

    View details for PubMedID 32764469

    View details for PubMedCentralID PMC7463940

  • Engineering the Bridge between Innate and Adaptive Immunity for Cancer Immunotherapy: Focus on gammadelta T and NK Cells. Cells Morandi, F., Yazdanifar, M., Cocco, C., Bertaina, A., Airoldi, I. 2020; 9 (8)


    Most studies on genetic engineering technologies for cancer immunotherapy based on allogeneic donors have focused on adaptive immunity. However, the main limitation of such approaches is that they can lead to severe graft-versus-host disease (GvHD). An alternative approach would bolster innate immunity by relying on the natural tropism of some subsets of the innate immune system, such as gammadelta T and natural killer (NK) cells, for the tumor microenvironment and their ability to kill in a major histocompatibility complex (MHC)-independent manner. gammadelta T and NK cells have the unique ability to bridge innate and adaptive immunity while responding to a broad range of tumors. Considering these properties, gammadelta T and NK cells represent ideal sources for developing allogeneic cell therapies. Recently, significant efforts have been made to exploit the intrinsic anti-tumor capacity of these cells for treating hematologic and solid malignancies using genetic engineering approaches such as chimeric antigen receptor (CAR) and T cell receptor (TCR). Here, we review over 30 studies on these two approaches that use gammadelta T and NK cells in adoptive cell therapy (ACT) for treating cancer. Based on those studies, we propose several promising strategies to optimize the clinical translation of these approaches.

    View details for DOI 10.3390/cells9081757

    View details for PubMedID 32707982

  • Gamma delta T cells as novel immunotherapeutic tool to cure drug-resistant viral infections in transplanted pediatric patients: development and validation of procedures for a new ATMP (advanced therapy medicinal product) Zorzoli, A., Morandi, F., Barbarito, G., Bertaina, A., Podesta, M., Airoldi, I. ELSEVIER SCI LTD. 2020: S139
  • CRISPR-based gene editing enables FOXP3 gene repair in IPEX patient cells. Science advances Goodwin, M., Lee, E., Lakshmanan, U., Shipp, S., Froessl, L., Barzaghi, F., Passerini, L., Narula, M., Sheikali, A., Lee, C. M., Bao, G., Bauer, C. S., Miller, H. K., Garcia-Lloret, M., Butte, M. J., Bertaina, A., Shah, A., Pavel-Dinu, M., Hendel, A., Porteus, M., Roncarolo, M. G., Bacchetta, R. 2020; 6 (19): eaaz0571


    The prototypical genetic autoimmune disease is immune dysregulation polyendocrinopathy enteropathy X-linked (IPEX) syndrome, a severe pediatric disease with limited treatment options. IPEX syndrome is caused by mutations in the forkhead box protein 3 (FOXP3) gene, which plays a critical role in immune regulation. As a monogenic disease, IPEX is an ideal candidate for a therapeutic approach in which autologous hematopoietic stem and progenitor (HSPC) cells or T cells are gene edited ex vivo and reinfused. Here, we describe a CRISPR-based gene correction permitting regulated expression of FOXP3 protein. We demonstrate that gene editing preserves HSPC differentiation potential, and that edited regulatory and effector T cells maintain their in vitro phenotype and function. Additionally, we show that this strategy is suitable for IPEX patient cells with diverse mutations. These results demonstrate the feasibility of gene correction, which will be instrumental for the development of therapeutic approaches for other genetic autoimmune diseases.

    View details for DOI 10.1126/sciadv.aaz0571

    View details for PubMedID 32494707

    View details for PubMedCentralID PMC7202871

  • CRISPR-based gene editing enables FOXP3 gene repair in IPEX patient cells SCIENCE ADVANCES Goodwin, M., Lee, E., Lakshmanan, U., Shipp, S., Froessl, L., Barzaghi, F., Passerini, L., Narula, M., Sheikali, A., Lee, C. M., Bao, G., Bauer, C. S., Miller, H. K., Garcia-Lloret, M., Butte, M. J., Bertaina, A., Shah, A., Pavel-Dinu, M., Hendel, A., Porteus, M., Roncarolo, M. G., Bacchetta, R. 2020; 6 (19)
  • Myeloablative conditioning for allo-HSCT in pediatric ALL: FTBI or chemotherapy?-A multicenter EBMT-PDWP study. Bone marrow transplantation Willasch, A. M., Peters, C., Sedlacek, P., Dalle, J., Kitra-Roussou, V., Yesilipek, A., Wachowiak, J., Lankester, A., Prete, A., Hamidieh, A. A., Ifversen, M., Buechner, J., Krivan, G., Hamladji, R., Diaz-de-Heredia, C., Skorobogatova, E., Michel, G., Locatelli, F., Bertaina, A., Veys, P., Dupont, S., Or, R., Gungor, T., Aleinikova, O., Sufliarska, S., Sundin, M., Rascon, J., Kaare, A., Nemet, D., Fagioli, F., Klingebiel, T. E., Styczynski, J., Bierings, M., Nagy, K., Abecasis, M., Afanasyev, B., Ansari, M., Vettenranta, K., Alseraihy, A., Chybicka, A., Robinson, S., Bertrand, Y., Kupesiz, A., Ghavamzadeh, A., Campos, A., Pichler, H., Dalissier, A., Labopin, M., Corbacioglu, S., Balduzzi, A., Galimard, J., Bader, P., EBMT Paediatric Diseases Working Party 2020


    Although most children with acute lymphoblastic leukemia (ALL) receive fractionated total body irradiation (FTBI) as myeloablative conditioning (MAC) for allogeneic hematopoietic stem cell transplantation (allo-HSCT), it is an important matter of debate if chemotherapy can effectively replace FTBI. To compare outcomes after FTBI versus chemotherapy-based conditioning (CC), we performed a retrospective EBMT registry study. Children aged 2-18 years after MAC for first allo-HSCT of bone marrow (BM) or peripheral blood stem cells (PBSC) from matched-related (MRD) or unrelated donors (UD) in first (CR1) or second remission (CR2) between 2000 and 2012 were included. Propensity score weighting was used to control pretreatment imbalances of the observed variables. 3.054 patients were analyzed. CR1 (1.498): median follow-up (FU) after FTBI (1.285) and CC (213) was 6.8 and 6.1 years. Survivals were not significantly different. CR2 (1.556): median FU after FTBI (1.345) and CC (211) was 6.2 years. Outcomes after FTBI were superior as compared with CC with regard to overall survival (OS), leukemia-free survival (LFS), relapse incidence (RI), and nonrelapse mortality (NRM). However, we must emphasize the preliminary character of the results of this retrospective "real-world-practice" study. These findings will be prospectively assessed in the ALL SCTped 2012 FORUM trial.

    View details for DOI 10.1038/s41409-020-0854-0

    View details for PubMedID 32203263

  • Treatment of steroid resistant acute graft versus host disease with an anti-CD26 monoclonal antibody-Begelomab. Bone marrow transplantation Bacigalupo, A., Angelucci, E., Raiola, A. M., Varaldo, R., Di Grazia, C., Gualandi, F., Benedetti, E., Risitano, A., Musso, M., Zallio, F., Ciceri, F., Chiusolo, P., Sica, S., Rambaldi, A., Bonifazi, F., Parma, M., Martino, M., Onida, F., Iori, A. P., Selleri, C., Borghero, C., Bertaina, A., Prezioso, L., Algeri, M., Locatelli, F. 2020


    We have treated 69 patients with steroid refractory acute graft versus host disease (SR-aGvHD), with an anti-CD26 monoclonal antibody (Begelomab): 28 patients in two prospective studies (EudraCT No. 2007-005809-21; EudraCT No. 2012-001353-19), and 41 patients on a compassionate use study. The median age of patients was 42 and 44 years; the severity of GvHD was as follows: grade II in 8 patients, grade III in 33, and grade IV in 28 patients. There were no adverse events directly attributable to the antibody. Day 28 response was 75% in the prospective studies and 61% in the compassionate use patients, with complete response rates of 11 and 12%. Response for grade III GvHD was 83 and 73% in the two groups; response in grade IV GvHD was 66 and 56% in the two groups. Non relapse mortality (NRM) at 6 months was 28 and 38%. Overall there were 64, 56, 68% responses for skin, liver, and gut stage 3-4 GvHD. The overall survival at 1 year was 50% for the prospective studies and 33% for the compassionate use patients. In conclusion, Begelomab induces over 60% responses in SR-aGvHD, including patients with severe gut and liver GvHD, having failed one or more lines of treatment.

    View details for DOI 10.1038/s41409-020-0855-z

    View details for PubMedID 32203257

  • A beta T-Cell/CD19 B-Cell Depleted Haploidentical Stem Cell Transplantation: A New Platform for Curing Rare and Monogenic Disorders Bertaina, A., Bacchetta, R., Lewis, D. B., Grimm, P. C., Shah, A. J., Agarwal, R., Concepcion, W., Czechowicz, A., Bhatia, N., Lahiri, P., Weinberg, K. I., Parkman, R., Porteus, M., Roncarolo, M. ELSEVIER SCIENCE INC. 2020: S288
  • Experience with Ruxolitinib (Jakafi (R)) As a Salvage Therapy for Graft-Versus-Host Disease in Children and Young Adults Mavers, M., Klinger, E., Shyr, D. C., Shah, A. J., Bertaina, A., Soni, S. ELSEVIER SCIENCE INC. 2020: S179
  • Regulatory Type 1 T Cell Infusion in Mismatched Related or Unrelated Hematopoietic Stem Cell Transplantation (HSCT) for Hematologic Malignancies Agarwal, R., Bacchetta, R., Bertaina, A., Chen, P., Saini, G., Shiraz, P., Bhatia, N., Roncarolo, M. ELSEVIER SCIENCE INC. 2020: S272–S273
  • Early Epigenetic Immune Quantification Following Alpha/Beta T-Cell/CD19 B-Cell Depleted Haploidentical Stem Cell Transplant Correlates with CD4+T Cell Recovery at Day+100 Mayers, M., Schulze, J., Barbarito, G., Lakshmanan, U., Parkman, R., Weinberg, K. I., Chu, J., Agarwal, R., Roncarolo, M., Sachsenmaier, C., Bacchetta, R., Bertaina, A. ELSEVIER SCIENCE INC. 2020: S305
  • γδ T Cells: The Ideal Tool for Cancer Immunotherapy. Cells Yazdanifar, M. n., Barbarito, G. n., Bertaina, A. n., Airoldi, I. n. 2020; 9 (5)


    γδ T cells have recently gained considerable attention as an attractive tool for cancer adoptive immunotherapy due to their potent anti-tumor activity and unique role in immunosurveillance. The remarkable success of engineered T cells for the treatment of hematological malignancies has revolutionized the field of adoptive cell immunotherapy. Accordingly, major efforts are underway to translate this exciting technology to the treatment of solid tumors and the development of allogeneic therapies. The unique features of γδ T cells, including their major histocompatibility complex (MHC)-independent anti-cancer activity, tissue tropism, and multivalent response against a broad spectrum of the tumors, render them ideal for designing universal 'third-party' cell products, with the potential to overcome the challenges of allogeneic cell therapy. In this review, we describe the crucial role of γδ T cells in anti-tumor immunosurveillance and we summarize the different approaches used for the ex vivo and in vivo expansion of γδ T cells suitable for the development of novel strategies for cancer therapy. We further discuss the different transduction strategies aiming at redirecting or improving the function of γδ T cells, as well as, the considerations for the clinical applications.

    View details for DOI 10.3390/cells9051305

    View details for PubMedID 32456316

  • Engineered type 1 regulatory T cells designed for clinical use kill primary pediatric acute myeloid leukemia cells Haematologica Cieniewicz, B., Uyeda, M. J., Chen, P. P., Sayitoglu, E. C., Liu, J. M., Andolfi, G., Greenthal, K., Bertaina, A., Gregori, S., Bacchetta, R., Lacayo, N. J., Cepika, A., Roncarolo, M. 2020
  • Alloantigen-specific Tr1 cells designed to prevent GvHD have a distinct molecular identity and suppress through CTLA-4 and PD-1 Society for Immunotherapy of Cancer’s (SITC) 35th Anniversary Annual Meeting Cepika, A., Chen, P. P., Uyeda, M. J., Cieniewicz, B., Narula, M., Amaya, L., Louis, D. M., Xu, L., Ji, X., Bertaina, A., Agarwal-Hashmi, R., Davis, M. M., Meyer, E., Bacchetta, R., Roncarolo, M. 2020: A159–A159
  • Genome editing of donor-derived T-cells to generate allogenic chimeric antigen receptor-modified T cells: Optimizing αβ T cell-depleted haploidentical hematopoietic stem cell transplantation. Haematologica Wiebking, V. n., Lee, C. M., Mostrel, N. n., Lahiri, P. n., Bak, R. n., Bao, G. n., Roncarolo, M. G., Bertaina, A. n., Porteus, M. H. 2020


    Allogeneic hematopoietic stem cell transplantation is an effective therapy for high-risk leukemias. In children, graft manipulation based on the selective removal of αβ T cells and B cells has been shown to reduce the risk of acute and chronic graft-versus-host disease, thus allowing the use of haploidentical donors which expands the population that allogeneic hematopoietic stem cell transplantation can be used in. Leukemic relapse, however, remains a problem. T cells expressing chimeric antigen receptors can potently eliminate leukemia, including in the central nervous system. We hypothesized that by modifying donor αβ T cells to simultaneously express a CD19-specific chimeric antigen receptors and inactivating the T cell receptor by genome editing, we could create a therapy that enhances the anti-leukemic efficacy of the stem cell transplant without increasing the risk of graft-versus-host disease. Using genome editing with Cas9 ribonucleoprotein and adeno-associated virus serotype 6, we integrate a CD19-specific chimeric antigen receptor in-frame into the TRAC locus. Greater than 90% of cells lost TCR expression, while >75% expressed the CAR. The product was further purified to ultimately have less than 0.05% residual TCR+ cells. In vitro, the CAR T cells efficiently eliminated target cells and produced high cytokine levels when challenged with CD19+ leukemia cells. In vivo, the gene modified T cells eliminated leukemia without causing xenogeneic graft-versus-host disease in a xenograft model. Gene editing was highly specific with no evidence of off-target effects. These data support the concept that the addition of αβ T cell-derived, genome edited T cells expressing CD19-specific chimeric antigen receptors could enhance the anti-leukemic efficacy of αβ T cell-depleted haploidentical hematopoietic stem cell transplantation without increasing the risk of graft-versus-host disease.

    View details for DOI 10.3324/haematol.2019.233882

    View details for PubMedID 32241852

  • Making a case for using γδ T cells against SARS-CoV-2. Critical reviews in microbiology Yazdanifar, M. n., Mashkour, N. n., Bertaina, A. n. 2020: 1–14


    Intensive worldwide efforts are underway to determine both the pathogenesis of SARS-CoV-2 infection and the immune responses in COVID-19 patients in order to develop effective therapeutics and vaccines. One type of cell that may contribute to these immune responses is the γδ T lymphocyte, which plays a key role in immunosurveillance of the mucosal and epithelial barriers by rapidly responding to pathogens. Although found in low numbers in blood, γδ T cells consist the majority of tissue-resident T cells and participate in the front line of the host immune defense. Previous studies have demonstrated the critical protective role of γδ T cells in immune responses to other respiratory viruses, including SARS-CoV-1. However, no studies have profoundly investigated these cells in COVID-19 patients to date. γδ T cells can be safely expanded in vivo using existing inexpensive FDA-approved drugs such as bisphosphonate, in order to test its protective immune response to SARS-CoV-2. To support this line of research, we review insights gained from previous coronavirus research, along with recent findings, discussing the potential role of γδ T cells in controlling SARS-CoV-2. We conclude by proposing several strategies to enhance γδ T cell's antiviral function, which may be used in developing therapies for COVID-19.

    View details for DOI 10.1080/1040841X.2020.1822279

    View details for PubMedID 33023358

  • Human-engineered Treg-like cells suppress FOXP3-deficient T cells but preserve adaptive immune responses in vivo. Clinical & translational immunology Sato, Y. n., Passerini, L. n., Piening, B. D., Uyeda, M. J., Goodwin, M. n., Gregori, S. n., Snyder, M. P., Bertaina, A. n., Roncarolo, M. G., Bacchetta, R. n. 2020; 9 (11): e1214


    Genetic or acquired defects in FOXP3+ regulatory T cells (Tregs) play a key role in many immune-mediated diseases including immune dysregulation polyendocrinopathy, enteropathy, X-linked (IPEX) syndrome. Previously, we demonstrated CD4+ T cells from healthy donors and IPEX patients can be converted into functional Treg-like cells by lentiviral transfer of FOXP3 (CD4LVFOXP3). These CD4LVFOXP3 cells have potent regulatory function, suggesting their potential as an innovative therapeutic. Here, we present molecular and preclinical in vivo data supporting CD4LVFOXP3 cell clinical progression.The molecular characterisation of CD4LVFOXP3 cells included flow cytometry, qPCR, RNA-seq and TCR-seq. The in vivo suppressive function of CD4LVFOXP3 cells was assessed in xenograft-versus-host disease (xeno-GvHD) and FOXP3-deficient IPEX-like humanised mouse models. The safety of CD4LVFOXP3 cells was evaluated using peripheral blood (PB) humanised (hu)- mice testing their impact on immune response against pathogens, and immune surveillance against tumor antigens.We demonstrate that the conversion of CD4+ T cells to CD4LVFOXP3 cells leads to specific transcriptional changes as compared to CD4+ T-cell transduction in the absence of FOXP3, including upregulation of Treg-related genes. Furthermore, we observe specific preservation of a polyclonal TCR repertoire during in vitro cell production. Both allogeneic and autologous CD4LVFOXP3 cells protect from xeno-GvHD after two sequential infusions of effector T cells. CD4LVFOXP3 cells prevent hyper-proliferation of CD4+ memory T cells in the FOXP3-deficient IPEX-like hu-mice. CD4LVFOXP3 cells do not impede in vivo expansion of antigen-primed T cells or tumor clearance in the PB hu-mice.These data support the clinical readiness of CD4LVFOXP3 cells to treat IPEX syndrome and other immune-mediated diseases caused by insufficient or dysfunctional FOXP3+ Tregs.

    View details for DOI 10.1002/cti2.1214

    View details for PubMedID 33304583

    View details for PubMedCentralID PMC7688376

  • Veno-occlusive Disease in HSCT Patients: Consensus-based Recommendations for Risk Assessment, Diagnosis, and Management by the GITMO Group. Transplantation Bonifazi, F. n., Sica, S. n., Angeletti, A. n., Marktel, S. n., Prete, A. n., Iori, A. P., Olivari, D. n., Rossetti, G. n., Bertaina, A. n., Botti, S. n., Busca, A. n., Carella, A. M., Cerretti, R. n., Gargiulo, G. n., Grassi, A. n., Gualandi, F. n., Irrera, G. n., Milone, G. n., Risitano, A. M., Santarone, S. n., Vassallo, E. n., Zecca, M. n., Ciceri, F. n., Pomponio, G. n. 2020


    Variation in clinical practice affects veno-occlusive disease (VOD) management, mainly in patients who undergo allogeneic hematopoietic stem cell transplantation (HSCT). Disputes about diagnostic criteria, treatment and prophylaxis, due to the lack of high-quality data, are at the base of this variability. With the aim of limiting inconsistency in clinical care, thus improving both patient outcomes and data collection reliability, the Italian Society of Stem cell transplant (GITMO) launched a collaborative effort to formulate recommendations based on integration of available evidence and expert's consensus. A systematic method, according to US National Institute of Health (NIH) guidelines and Italian National System for Guidelines, was used. Twenty-nine recommendations were approved with a strong (20) or weak (9) level of agreement, while 26 were rejected. In particular, the Panel pointed out the need to achieve an early diagnosis, encouraging the adoption of EBMT criteria and the prompt use of ultrasonography. Moreover, our experts strongly recommended in favour of prophylactic use of ursodeoxicolic acid (UDCA). As soon as a VOD diagnosis is established, treatment with defibrotide should be started for at least 21d. A number of areas of uncertainty, particularly concerning risk stratification and use of diagnostic tools such as elastography has been identified and discussed.

    View details for DOI 10.1097/TP.0000000000003569

    View details for PubMedID 33273315

  • High-Throughput RNA Sequencing Analysis Reveals Distinct Molecular Signature in NRAS and PTPN11 JMML Patients Leoncini, P., Silvestris, A., Vitullo, P., Zecca, M., Flotho, C., Niemeyer, C. M., Gallo, A., Bertaina, A., Locatelli, F., Merli, P. AMER SOC HEMATOLOGY. 2019
  • Results of a Multicentre, Randomized, Controlled Open-Label Study on the Use of Anti-T-Lymphocyte Globulin (ATLG) and Rituximab for Immunomodulation of Graft-Versus-Host Disease (GvHD) and Graft Failure (GF) in Patients with Non-Malignant Disorders Algeri, M., Galimberti, S., Bernardo, M., Rovelli, A., Zecca, M., La Nasa, G., Marktel, S., Merli, P., Bertaina, A., Pagliara, D., Boccieri, E., Del Bufalo, F., Gaspari, S., Capitoli, G., Valsecchi, M., Locatelli, F. AMER SOC HEMATOLOGY. 2019
  • Identification of Dual Positive CD19+/CD3+T Cells in an Apheresis Product Undergoing Chimeric Antigen Receptor (CAR) Transduction Schultz, L. M., Patel, S., Ramakrishna, S., Bertaina, A., Bhatia, N., Oak, J., Majzner, R. G., Baggott, C., Davis, K. L., Mackall, C. L., Feldman, S. A. AMER SOC HEMATOLOGY. 2019
  • Population pharmacokinetics of treosulfan in paediatric patients undergoing hematopoietic stem cell transplantation BRITISH JOURNAL OF CLINICAL PHARMACOLOGY van der Stoep, M. C., Zwaveling, J., Bertaina, A., Locatelli, F., Guchelaar, H. J., Lankester, A. C., Moes, D. R. 2019; 85 (9): 2033-2044


    Treosulfan is an alkylating agent increasingly used prior to haematopoietic stem cell transplantation. The aim of this study was to develop a population pharmacokinetic (PK) model of treosulfan in paediatric haematopoietic stem cell transplantation recipients and to explore the effect of potential covariates on treosulfan PK. Also, a limited sampling model (LSM) will be developed to accurately predict treosulfan exposure suitable for a therapeutic drug monitoring setting.In this multicentre study, 91 patients, receiving a total dose of 30, 36 or 42 g/m2 treosulfan, administered over 3 consecutive days, were enrolled. A population PK model was developed and demographic factors, as well as laboratory parameters, were included as potential covariates. In addition, a LSM was developed using data from 28 patients.A 2-compartment model with first order elimination best described the data. Bodyweight with allometric scaling and maturation function were identified as significant predictors of treosulfan clearance. Treosulfan clearance reaches 90% of adult values at 4 postnatal years. A model-based dosing table is presented to target an exposure of 1650 mg*h/L (population median) for different weight and age groups. Samples taken at 1.5, 4 and 7 hours after start of infusion resulted in the best limited sampling strategy.This study provides a treosulfan population PK model in children and captures the developmental changes in clearance. A 3-point LSM allows for accurate and precise estimation of treosulfan exposure.

    View details for DOI 10.1111/bcp.13995

    View details for Web of Science ID 000482997800018

    View details for PubMedID 31144349

    View details for PubMedCentralID PMC6710524

  • Graft Engineering and Adoptive Immunotherapy: New Approaches to Promote Immune Tolerance After Hematopoietic Stem Cell Transplantation FRONTIERS IN IMMUNOLOGY Bertaina, A., Roncarolo, M. 2019; 10
  • Myeloablative conditioning for first allogeneic HSCT in pediatric all: FTBI or chemotherapy? - An update of the retrospective multicenter EBMT-PDWP study Willasch, A., Peters, C., Sedlacek, P., Dalle, J., Kitra-Roussou, V., Yesilipek, A., Wachowiak, J., Lankester, A., Prete, A., Hamidieh, A., Ifversen, M., Buechner, J., Krivan, G., Hamladji, R., Diaz de Heredia, C., Skoroboga-Tova, E., Michel, G., Locatelli, F., Bertaina, A., Veys, P., Dupont, S., Or, R., Gungor, T., Aleinikova, O., Sufliarska, S., Sundin, M., Rascon, J., Kaare, A., Nemet, D., Fagioli, F., Klingebiel, T., Styczynski, J., Bierings, M., Nagy, K., Abecasis, M., Afanasyev, B., Ansari, M., Venntenranta, K., Alseraihy, A., Chybicka, A., Robinson, S., Bertrand, Y., Kupesiz, A., Ghavamzadeh, A., Campos, A., Dalissier, A., Labopin, M., Corbacioglu, S., Galimard, J., Bader, P. NATURE PUBLISHING GROUP. 2019: 137–38
  • genome editing of graft-derived T cells for post-transplant immunotherapy in combination with TCR alpha beta(+)/CD19(+)-depleted haploidentical HSCT Wiebking, V., Lee, C. M., Mostrel, N., Bak, R. O., Bao, G., Bertaina, A., Porteus, M. H. NATURE PUBLISHING GROUP. 2019: 183–84
  • Outcomes and Treatment Strategies for Autoimmunity and Hyperinflammation in Patients with RAG Deficiency JOURNAL OF ALLERGY AND CLINICAL IMMUNOLOGY-IN PRACTICE Farmer, J. R., Foldvari, Z., Ujhazi, B., De Ravin, S., Chen, K., Bleesing, J. H., Schuetz, C., Al-Herz, W., Abraham, R. S., Joshi, A. Y., Costa-Carvalho, B. T., Buchbinder, D., Booth, C., Reiff, A., Ferguson, P. J., Aghamohammadi, A., Abolhassani, H., Puck, J. M., Adeli, M., Cancrini, C., Palma, P., Bertaina, A., Locatelli, F., Di Matteo, G., Geha, R. S., Kanariou, M. G., Lycopoulou, L., Tzanoudaki, M., Sleasman, J. W., Parikh, S., Pinero, G., Fischer, B. M., Dbaibo, G., Unal, E., Patiroglu, T., Karakukcu, M., Al-Saad, K., Dilley, M. A., Pai, S., Dutmer, C. M., Gelfand, E. W., Geier, C. B., Eibl, M. M., Wolf, H. M., Henderson, L. A., Hazen, M. M., Bonfim, C., Wolska-Kusnierz, B., Butte, M. J., Hernandez, J. D., Nicholas, S. K., Stepensky, P., Chandrakasan, S., Miano, M., Westermann-Clark, E., Goda, V., Krivan, G., Holland, S. M., Fadugba, O., Henrickson, S. E., Ozen, A., Karakoc-Aydiner, E., Baris, S., Kiykim, A., Bredius, R., Hoeger, B., Boztug, K., Pashchenko, O., Neven, B., Moshous, D., de Villartay, J., Bousfiha, A., Hill, H. R., Notarangelo, L. D., Walter, J. E. 2019; 7 (6): 1970-+
  • Solid organ transplantation after hematopoietic stem cell transplantation in childhood: A multicentric retrospective survey AMERICAN JOURNAL OF TRANSPLANTATION Faraci, M., Bertaina, A., Dalissier, A., Ifversen, M., Schulz, A., Gennery, A., Burkhardt, B., Badell Serra, I., Diaz-de-Heredia, C., Lanino, E., Lankester, A. C., Gruhn, B., Matthes-Martin, S., Kuehl, J. S., Varotto, S., Paillard, C., Guilmatre, A., Sastre, A., Abecasis, M., Garwer, B., Sedlacek, P., Boelens, J. J., Beohou, E., Bader, P., EBMT Pediat Dis Working Party 2019; 19 (6): 1798–1805

    View details for DOI 10.1111/ajt.15240

    View details for Web of Science ID 000469275600024

  • Killer Ig-Like Receptors (KIRs): Their Role in NK Cell Modulation and Developments Leading to Their Clinical Exploitation FRONTIERS IN IMMUNOLOGY Pende, D., Falco, M., Vitale, M., Cantoni, C., Vitale, C., Munari, E., Bertaina, A., Moretta, F., Del Zotto, G., Pietra, G., Mingari, M., Locatelli, F., Moretta, L. 2019; 10
  • gamma delta T CELLS: A POTENT IMMUNOTHERAPY TOOL FOR CHILDREN WITH HIGH-RISK ACUTE LEUKEMIA Barbarito, G., Patil, R., Favaro, P., Sinha, R., Zorzoli, A., Klein, C. A., Airoldi, I., Bertaina, A. ELSEVIER SCI LTD. 2019: S31
  • Outcomes and treatment strategies for autoimmunity and hyperinflammation in patients with RAG deficiency. The journal of allergy and clinical immunology. In practice Farmer, J. R., Foldvari, Z., Ujhazi, B., De Ravin, S. S., Chen, K., Bleesing, J. J., Schuetz, C., Al-Herz, W., Abraham, R. S., Joshi, A. Y., Costa-Carvalho, B. T., Buchbinder, D., Booth, C., Reiff, A., Ferguson, P. J., Aghamohammadi, A., Abolhassani, H., Puck, J. M., Adeli, M., Cancrini, C., Palma, P., Bertaina, A., Locatelli, F., Di Matteo, G., Geha, R. S., Kanariou, M. G., Lycopoulou, E., Tzanoudaki, M., Sleasman, J. W., Parikh, S., Pinero, G., Fischer, B. M., Dbaibo, G., Unal, E., Patiroglu, T., Karakukcu, M., Al-Saad, K. K., Dilley, M. A., Pai, S., Dutmer, C. M., Gelfand, E. W., Geier, C. B., Eibl, M. M., Wolf, H. M., Henderson, L. A., Hazen, M. M., Bonfim, C., Wolska-Kusnierz, B., Butte, M. J., Hernandez, J. D., Nicholas, S. K., Stepensky, P., Chandrakasan, S., Miano, M., Westermann-Clark, E., Goda, V., Krivan, G., Holland, S. M., Fadugba, O., Henrickson, S. E., Ozen, A., Karakoc-Aydiner, E., Baris, S., Kiykim, A., Bredius, R., Hoeger, B., Boztug, K., Pashchenko, O., Neven, B., Moshous, D., Villartay, J. d., Bousfiha, A. A., Hill, H. R., Notarangelo, L. D., Walter, J. E. 2019


    BACKGROUND: While autoimmunity and hyperinflammation secondary to recombinase activating gene (RAG) deficiency have been associated with delayed diagnosis and even death, our current understanding is limited primarily to small case series.OBJECTIVE: Understand the frequency, severity, and treatment responsiveness of autoimmunity and hyperinflammation in RAG deficiency.METHODS: In reviewing the literature and our own database, we identified 85 patients with RAG deficiency, reported between 2001 and 2016, and compiled the largest case series to date of 63 patients with prominent autoimmune and/or hyperinflammatory pathology.RESULTS: Diagnosis of RAG deficiency was delayed a median of 5 years from the first clinical signs of immune dysregulation. The majority of patients (55.6%) presented with more than one autoimmune or hyperinflammatory complication, with the most common etiologies being cytopenias (84.1%), granulomas (23.8%), and inflammatory skin disorders (19.0%). Infections, including live viral vaccinations, closely preceded the onset of autoimmunity in 28.6% of cases. Autoimmune cytopenias had early onset (median 1.9, 2.1, and 2.6 years for autoimmune hemolytic anemia (AIHA), immune thrombocytopenia (ITP) and autoimmune neutropenia (AN), respectively) and were refractory to intravenous immunoglobulin, steroids, and rituximab in the majority of cases (64.7%, 73.7%, and 71.4% for AIHA, ITP, and AN, respectively). Evans syndrome specifically was associated with lack of response to first-line therapy. Treatment-refractory autoimmunity/hyperinflammation prompted hematopoietic stem cell transplantation in 20 patients.CONCLUSIONS: Autoimmunity/hyperinflammation can be a presenting sign of RAG deficiency and should prompt further evaluation. Multi-lineage cytopenias are often refractory to immunosuppressive treatment and may require hematopoietic cell transplantation for definitive management.

    View details for PubMedID 30877075

  • Chimerism Analysis in Pediatric Hematopoietic Stem Cell Transplantation for Non-Malignant Disorders Mariano, L., Zhang, B., Kristovich, K., Agarwal-Hashmi, R., Roncarolo, M., Bertaina, A., Fernandez-Vina, M. ELSEVIER SCIENCE INC. 2019
  • Outcome of Children Developing Grade III-IV Acute Graft-Versus-Host Disease after Allogeneic Hematopoietic Stem Cell Transplantation Bertaina, A., Locatelli, F., Galimard, J., Lawitschka, A., Balduzzi, A., Dalle, J., Sedlacek, P., Willasch, A., Yaniv, I., Labopin, M., Peters, C., Bader, P. ELSEVIER SCIENCE INC. 2019
  • Regulatory Type 1 T Cell Infusion in Mismatched Related or Unrelated Hematopoietic Stem Cell Transplantation (HSCT) for Hematologic Malignancies Agarwal, R., Bacchetta, R., Bertaina, A., Chu, J., Chen, P., Saini, G., Bhatia, N., Roncarolo, M. ELSEVIER SCIENCE INC. 2019
  • Epigenetic Immune Cell Quantification for Diagnosis and Monitoring of Patients with Primary Immune Deficiencies and Immune Regulatory Disorders Schulze, J., Werner, J., Schildknecht, K., Lakshmanan, U., Gruetzkau, A., Chu, J., Gernez, Y., Speckmann, C., Weinacht, K. G., Bertaina, A., Baron, U., Borte, S., Olek, S., Bacchetta, R. SPRINGER/PLENUM PUBLISHERS. 2019: S30
  • Haploidentical HSCT EBMT HANDBOOK: HEMATOPOIETIC STEM CELL TRANSPLANTATION AND CELLULAR THERAPIES Ciceri, F., Bacigalupo, A., Lankester, A., Bertaina, A., Carreras, E., Dufour, C., Mohty, M., Kroger, N. 2019: 479–86
  • Solid organ transplantation after haematopoietic stem cell transplantation in childhood: a multicentric retrospective survey. American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons Faraci, M., Bertaina, A., Dalissier, A., Ifversen, M., Schulz, A., Gennery, A., Burkhardt, B., Badell Serra, I., Diaz-de-Heredia, C., Lanino, E., Lankester, A. C., Gruhn, B., Matthes-Martin, S., Kuhl, J. S., Varotto, S., Paillard, C., Guilmatre, A., Sastre, A., Abecasis, M., Garwer, B., Sedlacek, P., Boelens, J. J., Beohou, E., Bader, P., EBMT Pediatric Disease Working Party 2018


    We report data obtained from a retrospective multicenter pediatric survey on behalf of the European Society for Blood and Marrow Transplantation (EBMT). Information on Solid Organ Transplantation (SOT) performed in pediatric recipients of either autologous or allogeneic hematopoietic stem cell transplantation (HSCT) between 1984 and 2016 were collected in 20 pediatric EBMT Centers (25.6%). Overall, we evaluated data on 44 SOTs following HSCT including 20 liver (LTx), 12 lung (LuTx), 6 heart (HTx), and 6 kidney (KTx) transplantations. The indication for SOT was organ failure related to intractable Graft-versus-Host Disease in 16 children (36.3%), acute or chronic HSCT-related toxicity in 18 (40.9%) and organ dysfunction related to the underlying disease in 10 (22.8%). The median follow-up was 10.9 years (95% CI: 1.7-29.5). The overall survival (OS) rate at 1 and 5 years after SOT was 85.7% and 80.4%, respectively: it was 74% and 63.2% after LTx, 83.2% after HTx, and 100% equally after LuTx and KTx. This multicenter survey confirms that SOT represents a promising option in children with severe organ failure occurred after HSCT. Additional studies are needed to further establish the effectiveness of SOT after HSCT and to better understand the mechanism underlying this encouraging success. This article is protected by copyright. All rights reserved.

    View details for PubMedID 30586230

  • T- and B-Cell Neogenesis Recovers Efficiently in Children with Acute Leukemia Given an Alpha-Beta-Cell Depleted Haplo-HSCT Followed By Infusion of Donor T-Cells Genetically Modified with Inducible Caspase 9 Suicide Gene (BPX-501 cells) Catanoso, M., Clave, E., Douay, C., Algeri, M., Fournier, I., Bertaina, V., Araujo, I., Del Bufalo, F., Galaverna, F., Surace, C., Bertaina, A., Merli, P., Foster, A., Toubert, A., Locatelli, F. AMER SOC HEMATOLOGY. 2018
  • Unrelated donor vs HLA-haploidentical alpha/beta T-cell and B-cell depleted HSCT in children with acute leukemia. Blood Bertaina, A., Zecca, M., Buldini, B., Sacchi, N., Algeri, M., Saglio, F., Perotti, C., Gallina, A. M., Bertaina, V., Lanino, E., Prete, A., Barberi, W., Tumino, M., Favre, C., Cesaro, S., Del Bufalo, F., Ripaldi, M., Boghen, S., Casazza, G., Rabusin, M., Balduzzi, A., Fagioli, F., Pagliara, D., Locatelli, F. 2018


    Traditionally, hematopoietic stem cell transplantation (HSCT) from both HLA-matched related and unrelated donors (UD) has been used for treating acute leukemia (AL) children in need of an allograft. Recently, HLA-haploidentical HSCT after alphabeta T-cell/B-cell depletion (alphabetahaplo-HSCT) was shown to be effective in single-center studies. Here, we report the first multicenter retrospective analysis of 127 macthed UD (MUD), 118 mismatched (MMUD) and 98 alphabetahaplo-HSCT recipients, transplanted between 2010 and 2015, in 13 Italian centers. All these AL children were transplanted in morphological remission after a myeloablative conditioning regimen. Graft failure occurred in 2% each of UD-HSCT and alphabetahaplo-HSCT group. In MUD vs MMUD-HSCT recipients, the cumulative incidence (CI) of grade II-IV and grade III-IV acute GvHD was 35% vs 44% and 6% vs 18%, as compared to 16% and 0% in alphabetahaplo-HSCT recipients (P<0.001). Children treated with alphabetahaplo-HSCT also had a significantly lower incidence of overall and extensive chronic GvHD (p<0.01). Eight (6%) MUD, 32 (28%) MMUD and 9 (9%) alphabetahaplo-HSCT patients died from transplant-related complications. With a median follow-up of 3.3 years, the 5-year probability of leukemia-free survival in the 3 groups was 67%, 55% and 62% respectively. In the three groups, chronic GvHD-free/relapse-free (GRFS) probability of survival was 61%, 34% and 58%, respectively (P<0.001). When compared to patients given MMUD-HSCT, alphabetahaplo-HSCT recipients had a lower CI of NRM and a better GFRS (P<0.001). These data indicate that alphabetahaplo-HSCT is a suitable therapeutic option for children with AL in need of transplantation, especially when an allele-matched UD is not available.

    View details for DOI 10.1182/blood-2018-07-861575

    View details for PubMedID 30348653

  • Analysis of KIR3DP1 Polymorphism Provides Relevant Information on Centromeric KIR Gene Content JOURNAL OF IMMUNOLOGY Bono, M., Pende, D., Bertaina, A., Moretta, A., Della Chiesa, M., Sivori, S., Zecca, M., Locatelli, F., Moretta, L., Bottino, C., Falco, M. 2018; 201 (5): 1460-1467


    Four killer cell Ig-like receptor (KIR) genes, collectively referred to as framework genes, characterize almost all KIR haplotypes. In particular, KIR3DL3 and KIR3DL2 mark the ends of the locus, whereas KIR3DP1 and KIR2DL4 are located in the central part. A recombination hot spot, mapped between KIR3DP1 and KIR2DL4, splits the haplotypes into two regions: a centromeric (Cen) region (spanning from KIR3DL3 to KIR3DP1) and a telomeric region (from KIR2DL4 to KIR3DL2), both varying in KIR gene content. In this study, we analyzed KIR3DP1 polymorphism in a cohort of 316 healthy, unrelated individuals. To this aim, we divided KIR3DP1 alleles into two groups by the use of a sequence-specific primer- PCR approach. Our data clearly indicated that KIR3DP1 alleles present on haplotypes carrying Cen-A or Cen-B1 regions differ from those having Cen-B2 motifs. Few donors (∼3%) made exceptions, and they were all, except one, characterized by uncommon haplotypes, including either KIR deletions or KIR duplications. Consequently, as KIR2DL1 is present in Cen-A and Cen-B1 regions but absent in Cen-B2 regions, we demonstrated that KIR3DP1 polymorphism might represent a suitable marker for KIR2DL1 gene copy number analysis. Moreover, because Cen-B1 and Cen-B2 regions are characterized by different KIR3DP1 alleles, we showed that KIR3DP1 polymorphism analysis also provides information to dissect between Cen-B1/Cen-B1 and Cen-B1/Cen-B2 donors. Taken together, our data suggest that the analysis of KIR3DP1 polymorphism should be included in KIR repertoire evaluation.

    View details for DOI 10.4049/jimmunol.1800564

    View details for Web of Science ID 000443584700015

    View details for PubMedID 30068594

  • Major Histocompatibility Complex and Hematopoietic Stem Cell Transplantation: Beyond the Classical HLA Polymorphism INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES Bertaina, A., Andreani, M. 2018; 19 (2)


    Allogeneic hematopoietic stem cell transplantation (HSCT) represents a curative treatment for many patients with hematological malignant or non-malignant disorders. Evaluation of potential donors for HSCT includes a rigorous assessment of the human leukocyte antigens (HLA) match status of family members, and the identification of suitable unrelated donors. Genes encoding transplantation antigens are placed both within and outside the major histocompatibility complex (MHC). The human MHC is located on the short arm of chromosome 6 and contains a series of genes encoding two distinct types of highly polymorphic cell surface glycoproteins. Donors for HSCT are routinely selected based on the level of matching for HLA-A, -B, -C, -DRB1, and -DQB1 loci. However, disease relapse, graft-versus-host-disease, and infection remain significant risk factors of morbidity and mortality. In the same breath, in high-risk patients, graft-versus-leukemia effects inherent in HLA mismatching play a substantial immunological role to limit the recurrence of post-transplant disease. The definition of a suitable donor is ever changing, shaped not only by current typing technology, but also by the specific transplant procedure. Indeed, a more complete understanding of permissible HLA mismatches and the role of Killer Immunoglobulin-like receptors' genes increases the availability of HLA-haploidentical and unrelated donors.

    View details for PubMedID 29470425

  • Voriconazole treatment in adults and children with hematological diseases: can it be used without measurement of plasma concentration? Medical mycology Girmenia, C. n., Annino, L. n., Bertaina, A. n., Mariotti, B. n., Caselli, D. n., Fanci, R. n., Barberi, W. n., Marchesi, F. n., Carotti, A. n., Ferrari, A. n., Cerchiara, E. n., Cupelli, L. n., Arcioni, F. n., Ribersani, M. n., Proia, A. n., Cartoni, C. n., Girardi, K. n., Venditti, A. n., Cassetta, M. I., Fallani, S. n., Novelli, A. n. 2018; 56 (3): 263–78


    Indication and timing of trough plasma-voriconazole (VCZ)-concentration (t-PVC) measurement during VCZ treatment is a debated issue. Patterns of t-PVC were prospectively evaluated in pediatric (50 courses) and adult (95 courses) hematologic patients. Efficacy patterns were defined: adequate, t-PVC always ≥1 mcg/ml; borderline, at least one t-PVC measurement <1 mcg/ml but median value of the measurements ≥1 mcg/ml; inadequate, median value of the measurements <1 mcg/ml. Toxicity patterns were defined: favorable, t-PVC always ≤5 mcg/ml; borderline, one or more t-PVC measurements >5 mcg/ml but median value of the measurements ≤5 mcg/ml; unfavorable, median value of the measurements >5 mcg/ml. In children and adults the mean t-PVCs were higher during intravenous treatments. The t-PVC efficacy pattern was adequate, borderline and inadequate in 48%, 12%, and 40% of courses, respectively, in children, and in 66.3%, 16.8%, and 16.8% of courses, respectively, in adults. Adequate efficacy pattern was more frequent in children with body weight above the median (≥25 kg) (OR 4.8; P = .011) and in adults with active hematological disease receiving intravenous therapy (OR 3.93; P = .006). Favorable toxicity pattern was more frequent in children receiving VCZ daily dosage below the median (<14 mg/kg) (OR 4.18; P = .027) and in adults with body weight below the median (<68 kg) (OR 0.22; P = .004). T-PVC measurement is generally needed, however, a non t-PVC guided approach may be considered in heavier adults receiving intravenous VCZ. The risk of supratherapeutic levels does not seem an absolute indication for t-PVC monitoring.

    View details for DOI 10.1093/mmy/myx053

    View details for PubMedID 28992093

  • High-Risk Leukemia: Past, Present, and Future Role of NK Cells JOURNAL OF IMMUNOLOGY RESEARCH Mavers, M., Bertaina, A. 2018: 1586905


    Natural killer (NK) cells are a population of cytotoxic innate lymphocytes that evolved prior to their adaptive counterparts and constitute one of the first lines of defense against infected/mutated cells. Several studies have shown that in patients with acute leukemia given haploidentical hematopoietic stem cell transplantation, donor-derived NK cells play a key role in the eradication of cancer cells. The antileukemic effect is mostly related to the presence of "alloreactive" NK cells, that is, mature KIR+ NK cells that express inhibitory KIR mismatched with HLA class I (KIR-L) of the patient. A genotypic analysis detecting KIR B haplotype and the relative B content is an additional donor selection criterion. These data provided the rationale for implementing phase I/II clinical trials of adoptive infusion of either selected or ex vivo-activated NK cells, often from an HLA-mismatched donor. In this review, we provide a historical perspective on the role played by NK cells in patients with acute leukemia, focusing also on the various approaches to adoptive NK cell therapy and the unresolved issues therein. In addition, we outline new methods to enhance NK activity, including anti-KIR monoclonal antibody, bi-/trispecific antibodies linking NK cells to cytokines and/or target antigens, and CAR-engineered NK cells.

    View details for PubMedID 29850617

    View details for PubMedCentralID PMC5925205

  • Hematopoietic stem cell transplantation for isolated extramedullary relapse of acute lymphoblastic leukemia in children. Bone marrow transplantation Gabelli, M. n., Zecca, M. n., Messina, C. n., Carraro, E. n., Buldini, B. n., Rovelli, A. M., Fagioli, F. n., Bertaina, A. n., Lanino, E. n., Favre, C. n., Rabusin, M. n., Prete, A. n., Ripaldi, M. n., Barberi, W. n., Porta, F. n., Caniglia, M. n., Santarone, S. n., D'Angelo, P. n., Basso, G. n., Locatelli, F. n. 2018


    Relapse of acute lymphoblastic leukemia (ALL) may occur in extramedullary sites, mainly central nervous system (CNS) and testis. Optimal post-remissional treatment for isolated extramedullary relapse (IEMR) is still controversial. We collected data of children treated with hematopoietic stem cell transplantation (HSCT) for ALL IEMR from 1990 to 2015 in Italy. Among 281 patients, 167 had a relapse confined to CNS, 73 to testis, 14 to mediastinum, and 27 to other organs. Ninety-seven patients underwent autologous HSCT, 79 received allogeneic HSCT from a matched family donor, 75 from a matched unrelated donor, and 30 from an HLA-haploidentical donor. The 10-year overall survival was 56% and was not influenced by gender, ALL blast immune-phenotype, age, site of relapse, duration of first remission, and type of HSCT. In multivariable analysis, the only prognostic factors were disease status at HSCT and year of transplantation. Patients transplanted in third or subsequent complete remission (CR) had a risk of death 2.3 times greater than those in CR2. Children treated after 2000 had half the risk of death than those treated before that year. Our results suggest that both autologous and allogeneic HSCT may be considered for the treatment of pediatric ALL IEMR after the achievement of CR2.

    View details for DOI 10.1038/s41409-018-0259-5

    View details for PubMedID 29899572

  • Late Development of FcεRγneg Adaptive Natural Killer Cells Upon Human Cytomegalovirus Reactivation in Umbilical Cord Blood Transplantation Recipients. Frontiers in immunology Muccio, L. n., Falco, M. n., Bertaina, A. n., Locatelli, F. n., Frassoni, F. n., Sivori, S. n., Moretta, L. n., Moretta, A. n., Della Chiesa, M. n. 2018; 9: 1050


    In human natural killer (NK) cells, human cytomegalovirus (HCMV) has been shown to be a driving force capable of inducing the expansion of a highly differentiated NKG2C+CD57+ subset, persisting over time in both HCMV+ healthy subjects and umbilical cord blood transplantation (UCBT) recipients experiencing HCMV viral reactivation. In HCMV+ healthy subjects, such expanded NK-cells are characterized by epigenetic modifications that modulate their phenotypic and functional characteristics. In particular, an enhanced ADCC activity is detectable in NK cells lacking the signaling protein FcεRγ. Timing and mechanisms involved in the acquisition of HCMV-induced, adaptive-like features by NK cells are currently unknown. In this study, we investigated the de novo acquisition of several adaptive features in NK cells developing after UCBT by monitoring NK-cell differentiation for at least 2 years after transplant. In UCBT recipients experiencing HCMV reactivation, a rapid phenotypic reconfiguration occurred resulting in the expected expansion of CD56dim NKG2C+CD57+ NK cells. However, while certain HCMV-driven adaptive hallmarks, including high KIR, LILRB1, CD2 and low/negative NKG2A, Siglec-7, and CD161 expression, were acquired early after UCBT (namely by month 6), downregulation of the signaling protein FcεRγ was detected at a later time interval (i.e., by month 12). This feature characterized only a minor fraction of the HCMV-imprinted NKG2C+CD57+ CD56dim NK cell subset, while it was detectable in higher proportions of CD57+ NK cells lacking NKG2C. Interestingly, in patients developing a hyporesponsive CD56-CD16bright NK-cell subset, FcεRγ downregulation occurred in these cells earlier than in CD56dim NK cells. Our data suggest that the acquisition of a fully "adaptive" profile requires signals that may lack in UCBT recipients and/or longer time is needed to obtain a stable epigenetic reprogramming. On the other hand, we found that both HCMV-induced FcεRγneg and FcεRγ+ NK cells from these patients, display similar CD107a degranulation and IFN-γ production capabilities in response to different stimuli, thus indicating that the acquisition of specialized effector functions can be achieved before the "adaptation" to HCMV is completed. Our study provides new insights in the process leading to the generation of different adaptive NK-cell subsets and may contribute to develop new approaches for their employment as novel immunotherapeutic tools.

    View details for DOI 10.3389/fimmu.2018.01050

    View details for PubMedID 29868012

    View details for PubMedCentralID PMC5968376

  • Dentate nucleus T1 hyperintensity: is it always gadolinium all that glitters? La Radiologia medica Pasquini, L. n., Rossi Espagnet, M. C., Napolitano, A. n., Longo, D. n., Bertaina, A. n., Visconti, E. n., Tomà, P. n. 2018; 123 (6): 469–73


    In the last few years, several scientific papers and reports have demonstrated magnetic resonance (MR) signal intensity (SI) changes on pre-contrast T1-weighted images following multiple gadolinium-based contrast agents (GBCA) administrations, particularly following the exposure to linear GBCAs. Pathological animal and human post-mortem studies have confirmed the relationship between this radiological finding and the presence of gadolinium accumulation in vulnerable brain regions in patients with normal renal function. In this short communication, we report the case of a 15-year-old patient affected by b-cell acute lymphoblastic leukemia (bALL) who developed a hyperintense signal in the dentate nuclei following multiple administrations of a macrocyclic GBCA. The purpose of this report is to discuss possible differential diagnoses of this radiological finding with special focus on the differentiation between iron or manganese accumulation, post-irradiation changes and GBCA-related Gd deposition, highlighting the importance of the acquisition of accurate clinical data to improve our scientific knowledge.

    View details for DOI 10.1007/s11547-017-0846-3

    View details for PubMedID 29374857

  • NG2 antigen is involved in leukemia invasiveness and central nervous system infiltration in MLL-rearranged infant B-ALL. Leukemia Prieto, C. n., López-Millán, B. n., Roca-Ho, H. n., Stam, R. W., Romero-Moya, D. n., Rodríguez-Baena, F. J., Sanjuan-Pla, A. n., Ayllón, V. n., Ramírez, M. n., Bardini, M. n., De Lorenzo, P. n., Valsecchi, M. G., Stanulla, M. n., Iglesias, M. n., Ballerini, P. n., Carcaboso, Á. M., Mora, J. n., Locatelli, F. n., Bertaina, A. n., Padilla, L. n., Carlos Rodríguez-Manzaneque, J. n., Bueno, C. n., Menéndez, P. n. 2018; 32 (3): 633–44


    Mixed-lineage leukemia (MLL)-rearranged (MLLr) infant B-cell acute lymphoblastic leukemia (iMLLr-B-ALL) has a dismal prognosis and is associated with a pro-B/mixed phenotype, therapy refractoriness and frequent central nervous system (CNS) disease/relapse. Neuron-glial antigen 2 (NG2) is specifically expressed in MLLr leukemias and is used in leukemia immunophenotyping because of its predictive value for MLLr acute leukemias. NG2 is involved in melanoma metastasis and brain development; however, its role in MLL-mediated leukemogenesis remains elusive. Here we evaluated whether NG2 distinguishes leukemia-initiating/propagating cells (L-ICs) and/or CNS-infiltrating cells (CNS-ICs) in iMLLr-B-ALL. Clinical data from the Interfant cohort of iMLLr-B-ALL demonstrated that high NG2 expression associates with lower event-free survival, higher number of circulating blasts and more frequent CNS disease/relapse. Serial xenotransplantation of primary MLL-AF4+ leukemias indicated that NG2 is a malleable marker that does not enrich for L-IC or CNS-IC in iMLLr-B-All. However, NG2 expression was highly upregulated in blasts infiltrating extramedullar hematopoietic sites and CNS, and specific blockage of NG2 resulted in almost complete loss of engraftment. Indeed, gene expression profiling of primary blasts and primografts revealed a migratory signature of NG2+ blasts. This study provides new insights on the biology of NG2 in iMLLr-B-ALL and suggests NG2 as a potential therapeutic target to reduce the risk of CNS disease/relapse and to provide safer CNS-directed therapies for iMLLr-B-ALL.

    View details for DOI 10.1038/leu.2017.294

    View details for PubMedID 28943635

    View details for PubMedCentralID PMC5843903

  • Phosphatidylinositol 3-kinase inhibition potentiates glucocorticoid response in B-cell acute lymphoblastic leukemia. Journal of cellular physiology Evangelisti, C. n., Cappellini, A. n., Oliveira, M. n., Fragoso, R. n., Barata, J. T., Bertaina, A. n., Locatelli, F. n., Simioni, C. n., Neri, L. M., Chiarini, F. n., Lonetti, A. n., Buontempo, F. n., Orsini, E. n., Pession, A. n., Manzoli, L. n., Martelli, A. M., Evangelisti, C. n. 2018; 233 (3): 1796–1811


    Despite remarkable progress in polychemotherapy protocols, pediatric B-cell acute lymphoblastic leukemia (B-ALL) remains fatal in around 20% of cases. Hence, novel targeted therapies are needed for patients with poor prognosis. Glucocorticoids (GCs) are drugs commonly administrated for B-ALL treatment. Activation of the phosphatidylinositol 3-kinase (PI3K)/Akt/mammalian target of rapamycin signaling pathway is frequently observed in B-ALL and contributes to GC-resistance. Here, we analyzed for the first time to our knowledge, the therapeutic potential of pan and isoform-selective PI3K p110 inhibitors, alone or combined with dexamethasone (DEX), in B-ALL leukemia cell lines and patient samples. We found that a pan PI3K p110 inhibitor displayed the most powerful cytotoxic effects in B-ALL cells, by inducing cell cycle arrest and apoptosis. Both a pan PI3K p110 inhibitor and a dual γ/δ PI3K p110 inhibitor sensitized B-ALL cells to DEX by restoring nuclear translocation of the GC receptor and counteracted stroma-induced DEX-resistance. Finally, gene expression analysis documented that, on one hand the combination consisting of a pan PI3K p110 inhibitor and DEX strengthened the DEX-induced up- or down-regulation of several genes involved in apoptosis, while on the other, it rescued the effects of genes that might be involved in GC-resistance. Overall, our findings strongly suggest that PI3K p110 inhibition could be a promising strategy for treating B-ALL patients by improving GC therapeutic effects and/or overcoming GC-resistance.

    View details for DOI 10.1002/jcp.26135

    View details for PubMedID 28777460

  • Outcome of children with acute leukemia given HLA-haploidentical HSCT after aß T-cell and B-cell depletion. Blood Locatelli, F., Merli, P., Pagliara, D., Li Pira, G., Falco, M., Pende, D., Rondelli, R., Lucarelli, B., Brescia, L. P., Masetti, R., Milano, G. M., Bertaina, V., Algeri, M., Pinto, R. M., Strocchio, L., Meazza, R., Grapulin, L., Handgretinger, R., Moretta, A., Bertaina, A., Moretta, L. 2017


    Allogeneic hematopoietic stem cell transplantation (HSCT) from an HLA-haploidentical relative (haplo-HSCT) is a suitable option for children with acute leukemia (AL) either relapsed or at high-risk of treatment failure. We developed a novel method of graft manipulation based on negative depletion of αβ T and B cells and conducted a prospective trial evaluating the outcome of children with AL transplanted with this approach. Eighty AL children, transplanted between September 2011 and September 2014, were enrolled in the trial. All children were given a fully myeloablative preparative regimen. Anti-T-lymphocyte globulin from day -5 to -3 was used for preventing graft rejection and graft-versus-host disease (GVHD); no patient received any posttransplantation GVHD prophylaxis. Two children experienced primary graft failure. The cumulative incidence of skin-only, grade 1-2 acute GVHD was 30%; no patient developed extensive chronic GVHD. Four patients died, the cumulative incidence of nonrelapse mortality being 5%, whereas 19 relapsed, resulting in a 24% cumulative incidence of relapse. With a median follow-up of 46 months for surviving patients, the 5-year probability of chronic GVHD-free, relapse-free survival (GRFS) is 71%. Total body irradiation-containing preparative regimen was the only variable favorably influencing relapse incidence and GRFS. The outcomes of these 80 patients are comparable to those of 41 and 51 children given transplantation from an HLA-identical sibling or a 10/10 allelic-matched unrelated donor in the same period. These data indicate that haplo-HSCT after αβ T- and B-cell depletion represents a competitive alternative for children with AL in need of urgent allograft. This trial was registered at as #NCT01810120.

    View details for DOI 10.1182/blood-2017-04-779769

    View details for PubMedID 28588018

  • Outcome of hematopoietic cell transplantation for DNA double-strand break repair disorders. journal of allergy and clinical immunology Slack, J., Albert, M. H., Balashov, D., Belohradsky, B. H., Bertaina, A., Bleesing, J., Booth, C., Buechner, J., Buckley, R. H., Ouachée-Chardin, M., Deripapa, E., Drabko, K., Eapen, M., Feuchtinger, T., Finocchi, A., Gaspar, H. B., Ghosh, S., Gillio, A., Gonzalez-Granado, L. I., Grunebaum, E., Güngör, T., Heilmann, C., Helminen, M., Higuchi, K., Imai, K., Kalwak, K., Kanazawa, N., Karasu, G., Kucuk, Z. Y., Laberko, A., Lange, A., Mahlaoui, N., Meisel, R., Moshous, D., Muramatsu, H., Parikh, S., Pasic, S., Schmid, I., Schuetz, C., Schulz, A., Schultz, K. R., Shaw, P. J., Slatter, M. A., Sykora, K., Tamura, S., Taskinen, M., Wawer, A., Wolska-Kus Nierz, B., Cowan, M. J., Fischer, A., Gennery, A. R. 2017


    Rare DNA breakage repair disorders predispose to infection and lymphoreticular malignancies. Hematopoietic cell transplantation (HCT) is curative, but coadministered chemotherapy or radiotherapy is damaging because of systemic radiosensitivity. We collected HCT outcome data for Nijmegen breakage syndrome, DNA ligase IV deficiency, Cernunnos-XRCC4-like factor (Cernunnos-XLF) deficiency, and ataxia-telangiectasia (AT).Data from 38 centers worldwide, including indication, donor, conditioning regimen, graft-versus-host disease, and outcome, were analyzed. Conditioning was classified as myeloablative conditioning (MAC) if it contained radiotherapy or alkylators and reduced-intensity conditioning (RIC) if no alkylators and/or 150 mg/m2 fludarabine or less and 40 mg/kg cyclophosphamide or less were used.Fifty-five new, 14 updated, and 18 previously published patients were analyzed. Median age at HCT was 48 months (range, 1.5-552 months). Twenty-nine patients underwent transplantation for infection, 21 had malignancy, 13 had bone marrow failure, 13 received pre-emptive transplantation, 5 had multiple indications, and 6 had no information. Twenty-two received MAC, 59 received RIC, and 4 were infused; information was unavailable for 2 patients. Seventy-three of 77 patients with DNA ligase IV deficiency, Cernunnos-XLF deficiency, or Nijmegen breakage syndrome received conditioning. Survival was 53 (69%) of 77 and was worse for those receiving MAC than for those receiving RIC (P = .006). Most deaths occurred early after transplantation, suggesting poor tolerance of conditioning. Survival in patients with AT was 25%. Forty-one (49%) of 83 patients experienced acute GvHD, which was less frequent in those receiving RIC compared with those receiving MAC (26/56 [46%] vs 12/21 [57%], P = .45). Median follow-up was 35 months (range, 2-168 months). No secondary malignancies were reported during 15 years of follow-up. Growth and developmental delay remained after HCT; immune-mediated complications resolved.RIC HCT resolves DNA repair disorder-associated immunodeficiency. Long-term follow-up is required for secondary malignancy surveillance. Routine HCT for AT is not recommended.

    View details for DOI 10.1016/j.jaci.2017.02.036

    View details for PubMedID 28392333

    View details for PubMedCentralID PMC5632132

  • T Cell-Depleted and T Cell-Replete HLA-Haploidentical Stem Cell Transplantation for Non-malignant Disorders CURRENT HEMATOLOGIC MALIGNANCY REPORTS Bertaina, A., Pitisci, A., Sinibaldi, M., Algeri, M. 2017; 12 (1): 68-78


    Hematopoietic stem cell transplantation (HSCT) is a treatment option for children with malignant and non-malignant disorders as well as an expanding number of inherited disorders. However, only a limited portion of patients in the need of an allograft have an HLA-compatible, either related or unrelated, donor. Haploidentical HSCT is now considered a valid treatment option, especially in view of the recent insights in terms of graft manipulation. This review will offer an overview of clinical results obtained through the use of haploidentical HSCT in non-malignant diseases. We will analyze major advantages and drawbacks of both T cell depleted and unmanipulated HSCT, discussing future challenges for further improving patients' outcome.T cell depletion (TCD) aims to reduce the morbidity and mortality associated with graft-versus-host disease (GvHD). However, the delayed immune recovery and the risk of graft failure still remain potential problems. In the last years, the use of post-transplant cyclophosphamide has been shown to be an alternative effective strategy to prevent GvHD in recipients of haploidentical HSCT. Recent data suggest that both T cell depleted and T cell-replete haplo-HSCT are suitable options to treat children with several types of non-malignant disorders lacking an HLA-identical donor.

    View details for DOI 10.1007/s11899-017-0364-3

    View details for Web of Science ID 000397936700009

    View details for PubMedID 28116633

  • The combination of bortezomib with chemotherapy to treatrelapsed/refractory acute lymphoblastic leukaemia of childhood BRITISH JOURNAL OF HAEMATOLOGY Bertaina, A., Vinti, L., Strocchio, L., Gaspari, S., Caruso, R., Algeri, M., Coletti, V., Gurnari, C., Romano, M., Cefalo, M. G., Girardi, K., Trevisan, V., Bertaina, V., Merli, P., Locatelli, F. 2017; 176 (4): 629-636


    Achieving complete remission (CR) in childhood relapsed/refractory acute lymphoblastic leukaemia (ALL) is a difficult task. Bortezomib, a proteasome inhibitor, has in vitro activity against ALL blasts. A phase I-II trial, reported by the Therapeutic Advances in Childhood Leukaemia and Lymphoma (TACL) consortium, demonstrated that bortezomib with chemotherapy has acceptable toxicity and remarkable activity in patients with relapsed ALL failing 2-3 previous regimens. We evaluated bortezomib in combination with chemotherapy in 30 and 7 children with B-cell precursor (BCP) and T-cell ALL, respectively. Bortezomib (1·3 mg/m2 /dose) was administered intravenously on days 1, 4, 8, and 11. Chemotherapy agents were the same as those used in the TACL trial, consisting of dexamethasone, doxorubicin, vincristine and pegylated asparaginase. Three patients (8·1%) died due to infections. Twenty-seven patients (72·9%) achieved CR or CR with incomplete platelet recovery (CRp). Fourteen had minimal residual disease (MRD) lower than 0·1%. Twenty-two of 30 BCP-ALL patients (73·3%) and 5/7 patients (71%) with T-cell ALL achieved CR/CRp. The 2-year overall survival (OS) is 31·3%; CR/CRp patients with an MRD response had a remarkable 2-year OS of 68·4%. These data confirm that the combination of bortezomib with chemotherapy is a suitable/effective option for childhood relapsed/refractory ALL.

    View details for DOI 10.1111/bjh.14505

    View details for Web of Science ID 000395081200014

    View details for PubMedID 28116786

  • Remestemcel-L for the treatment of graft versus host disease EXPERT REVIEW OF CLINICAL IMMUNOLOGY Locatelli, F., Algeri, M., Trevisan, V., Bertaina, A. 2017; 13 (1): 43-56


    Remestemcel-L, a third-party, off-the-shelf preparation of bone-marrow derived mesenchymal stromal cells (MSCs), has been developed for experimental use in acute graft-versus-host disease (aGvHD) and other immune-mediated conditions. Several preclinical and clinical studies have indeed suggested the potential of human mesenchymal stromal cells (MSCs) as an effective treatment for steroid-refractory aGvHD. However, an unambiguous demonstration of efficacy is still lacking. Areas covered: This review critically examines the biologic rationale supporting MSCs use in aGvHD and analyzes the results of published clinical trials in this setting, with a particular focus on the potential benefits and drawbacks of Remestemcel-L. For this purpose, a systematic literature search was performed in PubMed using the following keywords: 'mesenchymal stromal cells', 'mesenchymal progenitor cells', 'multipotent stromal cells', 'mesenchymal cells', 'MSC', 'Remestemcel-L', 'Prochymal', and 'graft-versus-host disease' or 'GvHD'. Expert commentary: Remestemcel-L represents a promising alternative to second-line immunosuppressive agents for the treatment of steroid-refractory aGvHD. Despite the safety and the favorable risk/benefit profile of this cell product, which has been demonstrated in several phase I-II studies, large and prospective randomized trials are required to confirm its efficacy in aGvHD and to define the optimal schedule of administration in terms of infusion timing, cell dose and pharmacological synergism.

    View details for DOI 10.1080/1744666X.2016.1208086

    View details for Web of Science ID 000390747200007

    View details for PubMedID 27399600

  • Novel X-Linked Inhibitor of Apoptosis Mutation in Very Early-Onset Inflammatory Bowel Disease Child Successfully Treated with HLA-Haploidentical Hemapoietic Stem Cells Transplant after Removal of αβ+ T and B Cells. Frontiers in immunology Cifaldi, C. n., Chiriaco, M. n., Di Matteo, G. n., Di Cesare, S. n., Alessia, S. n., De Angelis, P. n., Rea, F. n., Angelino, G. n., Pastore, M. n., Ferradini, V. n., Pagliara, D. n., Cancrini, C. n., Rossi, P. n., Bertaina, A. n., Finocchi, A. n. 2017; 8: 1893


    Monogenic defects in genes related to primary immunodeficiencies can be responsible for inflammatory bowel disease (IBD). Mutations in the X-linked inhibitor of apoptosis (XIAP) gene have been described in several patients suffering from IBD and, in particular, with very early-onset inflammatory bowel disease (VEOIBD) features. We report a VEOIBD child with a novel XIAP gene mutation characterized by a complicated disease course, which is unresponsive to several medical treatment options. A next-generation sequencing was performed and revealed a de novo hemizygous mutation in XIAP gene: c.565T>C p.L189P. After mutation discovery, we investigated the XIAP protein expression and nucleotide-binding oligomerization domain protein 2 (NOD2) signaling by western blotting. Flow-cytometry was used to analyze intracellular protein expression in different cell subsets and T cell apoptosis. We observed reduced protein expression in lymphocytes, granulocytes, monocytes, an Epstein-Barr virus-immortalized B cell line as well as increased apoptosis, and impairment in NOD2 signaling. The child was successfully treated with HLA-haploidentical hemapoietic stem cells transplant, acquired from his mother, after ex vivo elimination of α/β T cells and CD19 B cells. One year after the transplant, we repeated the analysis to appreciate the changes in his impairments. The recovery of XIAP protein expression, function, and normalization of apoptosis were observed. Our report emphasizes the important role of genetic analysis in the diagnosis of VEOIBD, illustrates the complete immunological and gastrointestinal recovery after transplant, and shows one of the few successful transplant cases of XIAP patients.

    View details for DOI 10.3389/fimmu.2017.01893

    View details for PubMedID 29312354

    View details for PubMedCentralID PMC5743702

  • Rabbit anti-human T-lymphocyte globulin and hematopoietic transplantation. Oncotarget Locatelli, F. n., Merli, P. n., Bertaina, A. n. 2017; 8 (57): 96460–61

    View details for DOI 10.18632/oncotarget.20878

    View details for PubMedID 29228536

    View details for PubMedCentralID PMC5722488

  • High interpatient variability of treosulfan exposure is associated with early toxicity in paediatric HSCT: a prospective multicentre study. British journal of haematology van der Stoep, M. Y., Bertaina, A. n., Ten Brink, M. H., Bredius, R. G., Smiers, F. J., Wanders, D. C., Moes, D. J., Locatelli, F. n., Guchelaar, H. J., Zwaveling, J. n., Lankester, A. C. 2017; 179 (5): 772–80


    Treosulfan-based conditioning is increasingly employed in paediatric haematopoietic stem cell transplantation (HSCT). Data on treosulfan pharmacokinetics in children are scarce, and the relationship between treosulfan exposure, toxicity and clinical outcome is unresolved. In this multicentre prospective observational study, we studied treosulfan pharmacokinetics and the drug's relationship with regimen-related toxicity and early clinical outcome in 77 paediatric patients. Treosulfan dose was 30 g/m2 , administered over 3 consecutive days in infants <1 year old (n = 12) and 42 g/m2 in children ≥1 year old (n = 65). Mean day 1 treosulfan exposure was 1744 ± 795 mg*h/l (10 g/m2 ) and 1561 ± 511 mg*h/l (14 g/m2 ), with an inter-individual variability of 56 and 33% in the respective groups. High treosulfan exposure (>1650 mg*h/l) was associated with an increased risk of mucosal [Odds ratio (OR) 4·40; 95% confidence interval (CI) 1·19-16·28, P = 0·026] and skin toxicity (OR 4·51; 95% CI 1·07-18·93, P = 0·040). No correlation was found between treosulfan exposure and the early clinical outcome parameters: engraftment, acute graft-versus-host disease and donor chimerism. Our study provides the first evidence in a large cohort of paediatric patients of high variability in treosulfan pharmacokinetics and an association between treosulfan exposure and early toxicity. Ongoing studies will reveal whether treosulfan exposure is related to long-term disease-specific outcome and late treatment-related toxicity.

    View details for DOI 10.1111/bjh.14960

    View details for PubMedID 29048102

  • Incidence, Risk Factors and Outcome of Pre-engraftment Gram-Negative Bacteremia After Allogeneic and Autologous Hematopoietic Stem Cell Transplantation: An Italian Prospective Multicenter Survey. Clinical infectious diseases : an official publication of the Infectious Diseases Society of America Girmenia, C. n., Bertaina, A. n., Piciocchi, A. n., Perruccio, K. n., Algarotti, A. n., Busca, A. n., Cattaneo, C. n., Raiola, A. M., Guidi, S. n., Iori, A. P., Candoni, A. n., Irrera, G. n., Milone, G. n., Marcacci, G. n., Scimè, R. n., Musso, M. n., Cudillo, L. n., Sica, S. n., Castagna, L. n., Corradini, P. n., Marchesi, F. n., Pastore, D. n., Alessandrino, E. P., Annaloro, C. n., Ciceri, F. n., Santarone, S. n., Nassi, L. n., Farina, C. n., Viscoli, C. n., Rossolini, G. M., Bonifazi, F. n., Rambaldi, A. n. 2017; 65 (11): 1884–96


    Gram-negative bacteremia (GNB) is a major cause of illness and death after hematopoietic stem cell transplantation (HSCT), and updated epidemiological investigation is advisable.We prospectively evaluated the epidemiology of pre-engraftment GNB in 1118 allogeneic HSCTs (allo-HSCTs) and 1625 autologous HSCTs (auto-HSCTs) among 54 transplant centers during 2014 (SIGNB-GITMO-AMCLI study). Using logistic regression methods. we identified risk factors for GNB and evaluated the impact of GNB on the 4-month overall-survival after transplant.The cumulative incidence of pre-engraftment GNB was 17.3% in allo-HSCT and 9% in auto-HSCT. Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa were the most common isolates. By multivariate analysis, variables associated with GNB were a diagnosis of acute leukemia, a transplant from a HLA-mismatched donor and from cord blood, older age, and duration of severe neutropenia in allo-HSCT, and a diagnosis of lymphoma, older age, and no antibacterial prophylaxis in auto-HSCT. A pretransplant infection by a resistant pathogen was significantly associated with an increased risk of posttransplant infection by the same microorganism in allo-HSCT. Colonization by resistant gram-negative bacteria was significantly associated with an increased rate of infection by the same pathogen in both transplant procedures. GNB was independently associated with increased mortality at 4 months both in allo-HSCT (hazard ratio, 2.13; 95% confidence interval, 1.45-3.13; P <.001) and auto-HSCT (2.43; 1.22-4.84; P = .01).Pre-engraftment GNB is an independent factor associated with increased mortality rate at 4 months after auto-HSCT and allo-HSCT. Previous infectious history and colonization monitoring represent major indicators of GNB.NCT02088840.

    View details for DOI 10.1093/cid/cix690

    View details for PubMedID 29020286

  • cell-depleted grafts from an HLA-haplo-identical donor. Oncoimmunology Bertaina, A., ZORZOLI, A., Petretto, A., Barbarito, G., Inglese, E., MERLI, P., LAVARELLO, C., Brescia, L. P., De Angelis, B., Tripodi, G., Moretta, L., Locatelli, F., Airoldi, I. 2017; 6 (2)


    We demonstrated that γδ T cells of patients given HLA-haploidentical HSCT after removal of αβ+ T cells and CD19+ B cells are endowed with the capacity of killing leukemia cells after ex vivo treatment with zoledronic acid (ZOL). Thus, we tested the hypothesis that infusion of ZOL in patients receiving this type of graft may enhance γδ T-cell cytotoxic activity against leukemia cells. ZOL was infused every 28 d in 43 patients; most were treated at least twice. γδ T cells before and after ZOL treatments were studied in 33 of these 43 patients, till at least 7 mo after HSCT by high-resolution mass spectrometry, flow-cytometry, and degranulation assay. An induction of Vδ2-cell differentiation, paralleled by increased cytotoxicity of both Vδ1 and Vδ2 cells against primary leukemia blasts was associated with ZOL treatment. Cytotoxic activity was further increased in Vδ2 cells, but not in Vδ1 lymphocytes in those patients given more than one treatment. Proteomic analysis of γδ T cells purified from patients showed upregulation of proteins involved in activation processes and immune response, paralleled by downregulation of proteins involved in proliferation. Moreover, a proteomic signature was identified for each ZOL treatment. Patients given three or more ZOL infusions had a better probability of survival in comparison to those given one or two treatments (86% vs. 54%, respectively, p = 0.008). Our data indicate that ZOL infusion in pediatric recipients of αβ T- and B-cell-depleted HLA-haploidentical HSCT promotes γδ T-cell differentiation and cytotoxicity and may influence the outcome of patients.

    View details for DOI 10.1080/2162402X.2016.1216291

    View details for PubMedID 28344861

    View details for PubMedCentralID PMC5353934

  • Whole Genome MBD-seq reveals different CpG methylation patterns in Azacytidine-treated Juvenile Myelomonocytic Leukaemia (JMML) patients. British journal of haematology Leoncini, P. P., Vitullo, P. n., Di Florio, F. n., Tocco, V. n., Cefalo, M. G., Pitisci, A. n., Girardi, K. n., Niemeyer, C. n., Locatelli, F. n., Bertaina, A. n. 2017

    View details for DOI 10.1111/bjh.14876

    View details for PubMedID 28771681

  • Efficacy of two different doses of rabbit anti-T-lymphocyte globulin to prevent graft-versus-host disease in children with haematological malignancies transplanted from an unrelated donor: a multicentre, randomised, open-label, phase 3 trial. The Lancet. Oncology Locatelli, F. n., Bernardo, M. E., Bertaina, A. n., Rognoni, C. n., Comoli, P. n., Rovelli, A. n., Pession, A. n., Fagioli, F. n., Favre, C. n., Lanino, E. n., Giorgiani, G. n., Merli, P. n., Pagliara, D. n., Prete, A. n., Zecca, M. n. 2017; 18 (8): 1126–36


    Although rabbit anti-T-lymphocyte globulin (ATLG) is largely used for the prevention of immune-mediated complications in patients given allogeneic haemopoietic stem-cell transplantation (HSCT) from an unrelated donor, the optimum dose of this drug in children is still undefined. We aimed to test whether a higher dose of ATLG was superior to a lower dose for prevention of grade II-IV acute graft-versus-host disease (GVHD).We conducted a multicentre, randomised, open-label, phase 3 trial in seven Italian centres comparing two different doses of ATLG (30 mg/kg vs 15 mg/kg, given intravenously over 3 days, from day -4 to -2) in children (aged 0-18 years) with haematological malignancies transplanted from an unrelated donor, selected using high-resolution typing for HLA-class I/II loci. All patients received a myeloablative regimen and cyclosporine-A plus short-term methotrexate as post-transplantation GVHD prophylaxis. Patients were randomly assigned (1:1) to either of the two groups and were stratified by the degree of HLA-compatibility with their donor, the source of haemopoietic stem cells used (bone marrow vs peripheral blood stem cells), and the disease risk category. The randomisation was open label; all investigators were aware of the treatment allocation. The primary endpoint of the study was 100-day cumulative incidence of grade II-IV acute GVHD. Statistical analyses were done according to the per-protocol principle. Other outcomes included cumulative incidence of chronic GVHD, non-relapse mortality, disease recurrence, and probability of overall survival and event-free survival. This study was registered with, number NCT00934557.Between Jan 15, 2008, and Sept 25, 2012, 89 patients were randomly assigned to the 30 mg/kg ATLG group and 91 to the 15 mg/kg ATLG group; 84 patients in the 30 mg/kg ATLG group and 88 in the 15 mg/kg ATLG group were included in the analysis. The median follow-up for the whole study population was 3·4 years (IQR 1·7-5·1). The 100-day cumulative incidence of grade II-IV acute GVHD was 36% (95% CI 28-48) in the 15 mg/kg ATLG group and 29% (20-40) in the 30 mg/kg ATLG group (hazard ratio [HR] 0·74, 95% CI 0·44-1·25; p=0·26). The cumulative incidence of non-relapse mortality was 9% (5-18) in the 15 mg/kg ATLG group and 19% (12-30) in the 30 mg/kg ATLG group (HR 2·08, 0·89-4·96; p=0·092). Cumulative incidence of disease recurrence was 15% (12-24): 14% (8-23) in the 15 mg/kg ATLG group versus 20% (13-31) in the 30 mg/kg ATLG group (HR 1·54, 0·74-3·21; p=0·25). The 5-year overall survival probability was 70% (62-77) for the whole study population: 78% (69-87) in the 15 mg/kg ATLG group versus 62% (50-73) in the 30 mg/kg ATLG group (HR 1·80, 1·01-3·20; p=0·045). The 5-year event-free survival was 77% for children in the 15 mg/kg ATLG group versus 61% in the 30 mg/kg ATLG group (HR 1·87, 1·07-3·28; p=0·028).Children with haematological malignancies transplanted from unrelated donors selected through high-resolution HLA-typing benefit from the use of a 15 mg/kg ATLG dose in comparison with a 30 mg/kg ATLG dose. ATLG at 15 mg/kg should thus be regarded as the standard serotherapy regimen for unrelated donor allogeneic HSCT in this patient population. Future randomised studies will continue to aim to optimise patient outcome and strategies to prevent acute GVHD occurrence.Fresenius/Neovii Biotech.

    View details for DOI 10.1016/S1470-2045(17)30417-5

    View details for PubMedID 28705454

  • Preservation of Antigen-Specific Functions of aß T Cells and B Cells Removed from Hematopoietic Stem Cell Transplants Suggests Their Use As an Alternative Cell Source for Advanced Manipulation and Adoptive Immunotherapy. Frontiers in immunology Li Pira, G., Di Cecca, S., Biagini, S., Girolami, E., Cicchetti, E., Bertaina, V., Quintarelli, C., Caruana, I., Lucarelli, B., Merli, P., Pagliara, D., Brescia, L. P., Bertaina, A., Montanari, M., Locatelli, F. 2017; 8: 332-?


    Hematopoietic stem cell transplantation is standard therapy for numerous hematological diseases. The use of haploidentical donors, sharing half of the HLA alleles with the recipient, has facilitated the use of this procedure as patients can rely on availability of a haploidentical donor within their family. Since HLA disparity increases the risk of graft-versus-host disease, T-cell depletion has been used to remove alloreactive lymphocytes from the graft. Selective removal of αβ T cells, which encompass the alloreactive repertoire, combined with removal of B cells to prevent EBV-related lymphoproliferative disease, proved safe and effective in clinical studies. Depleted αβ T cells and B cells are generally discarded as by-products. Considering the possible use of donor T cells for donor lymphocyte infusions or for generation of pathogen-specific T cells as mediators of graft-versus-infection effect, we tested whether cells in the discarded fractions were functionally intact. Response to alloantigens and to viral antigens comparable to that of unmanipulated cells indicated a functional integrity of αβ T cells, in spite of the manipulation used for their depletion. Furthermore, B cells proved to be efficient antigen-presenting cells, indicating that antigen uptake, processing, and presentation were fully preserved. Therefore, we propose that separated αβ T lymphocytes could be employed for obtaining pathogen-specific T cells, applying available methods for positive selection, which eventually leads to indirect allodepletion. In addition, these functional T cells could undergo additional manipulation, such as direct allodepletion or genetic modification.

    View details for DOI 10.3389/fimmu.2017.00332

    View details for PubMedID 28386262

  • Fungal infections of the lung in children PEDIATRIC RADIOLOGY Toma, P., Bertaina, A., Castagnola, E., Colafati, G. S., D'Andrea, M. L., Finocchi, A., Lucidi, V., Mastronuzzi, A., Granata, C. 2016; 46 (13): 1856-1865


    Fungal infections of the lungs are relatively common and potentially life-threatening conditions in immunocompromised children. The role of imaging in children with lung mycosis is to delineate the extension of pulmonary involvement, to assess response to therapy, and to monitor for adverse sequelae such as bronchiectasis and cavitation. The aim of this paper is to show imaging findings in a series of patients with fungal pneumonia from two tertiary children's hospitals, to discuss differential diagnoses and to show how imaging findings can vary depending on the host immune response.

    View details for DOI 10.1007/s00247-016-3696-6

    View details for Web of Science ID 000388743000012

    View details for PubMedID 27663906

  • Cytomegalovirus in hematopoietic stem cell transplant recipients - management of infection EXPERT REVIEW OF HEMATOLOGY Locatelli, F., Bertaina, A., Bertaina, V., Merli, P. 2016; 9 (11): 1093-1105


    Cytomegalovirus (CMV) still causes significant morbidity and mortality in patients given allogeneic hematopoietic stem cell transplantation (HSCT). Despite effective pharmacotherapy, potentially life-threatening CMV disease occurs nowadays in up to 10% of HSCT recipients; moreover, routinely used anti-CMV agents have been shown to be associated with morbidity. Areas covered: This review examines different issues related to diagnosis and management of CMV infection in HSCT recipients, paying particular attention to the monitoring of CMV-specific immune recovery, approaches of adoptive cell therapy and new antiviral drugs. Expert commentary: Despite advances in diagnostic tests and treatment, there is still room for refining management of CMV in HSCT recipients. Immunological monitoring should be associated in the future to virological monitoring. The safety profile and efficacy of new anti-CMV agents should be compared with that of standard-of-care drugs. Donor-derived, pathogen-specific T cells adoptively transferred after transplantation could contribute to reduce the impact of CMV infection on patient's outcome.

    View details for DOI 10.1080/17474086.2016.1242406

    View details for Web of Science ID 000387510600010

    View details for PubMedID 27690683

  • Selective Depletion of alpha beta T Cells and B Cells for Human Leukocyte Antigen-Haploidentical Hematopoietic Stem Cell Transplantation. A Three-Year Follow-Up of Procedure Efficiency BIOLOGY OF BLOOD AND MARROW TRANSPLANTATION Li Pira, G., Malaspina, D., Girolami, E., Biagini, S., Cicchetti, E., Conflitti, G., Broglia, M., Ceccarelli, S., Lazzaro, S., Pagliara, D., Meschini, A., Bertaina, A., Montanari, M., Locatelli, F. 2016; 22 (11): 2056-2064


    HLA-haploidentical family donors represent a valuable option for children requiring allogeneic hematopoietic stem cell transplantation (HSCT). Because graft-versus-host diseases (GVHD) is a major complication of HLA-haploidentical HSCT because of alloreactive T cells in the graft, different methods have been used for ex vivo T cell depletion. Removal of donor αβ T cells, the subset responsible for GVHD, and of B cells, responsible for post-transplantation lymphoproliferative disorders, have been recently developed for HLA-haploidentical HSCT. This manipulation preserves, in addition to CD34+ progenitors, natural killer, γδ T, and monocytes/dendritic cells, contributing to anti-leukemia activity and protection against infections. We analyzed depletion efficiency and cell yield in 200 procedures performed in the last 3 years at our center. Donors underwent CD34+  hematopoietic stem cell (HSC) peripheral blood mobilization with granulocyte colony-stimulating factor (G-CSF). Poor CD34+ cell mobilizers (48 of 189, 25%) received plerixafor in addition to G-CSF. Aphereses containing a median of 52.5 × 109 nucleated cells and 494 × 106 CD34+ HSC were manipulated using the CliniMACS device. In comparison to the initial product, αβ T cell depletion produced a median 4.1-log reduction (range, 3.1 to 5.5) and B cell depletion led to a median 3.4-log reduction (range, 2.0 to 4.7). Graft products contained a median of 18.5 × 106 CD34+ HSC/kg recipient body weight, with median values of residual αβ T cells and B cells of 29 × 103/kg and 33 × 103/kg, respectively. Depletion efficiency monitored at 6-month intervals demonstrated steady performance, while improved recovery of CD34+ cells was observed after the first year (P = .0005). These data indicate that αβ T cell and B cell depletion of HSC grafts from HLA-haploidentical donors was efficient and reproducible.

    View details for DOI 10.1016/j.bbmt.2016.08.006

    View details for Web of Science ID 000386544200021

    View details for PubMedID 27519279

  • Improving nelarabine efficacy in T cell acute lymphoblastic leukemia by targeting aberrant PI3K/AKT/mTOR signaling pathway JOURNAL OF HEMATOLOGY & ONCOLOGY Lonetti, A., Cappellini, A., Bertaina, A., Locatelli, F., Pession, A., Buontempo, F., Evangelisti, C., Evangelisti, C., Orsini, E., Zambonin, L., Neri, L. M., Martelli, A. M., Chiarini, F. 2016; 9


    Although in recent years, the introduction of novel chemotherapy protocols has improved the outcome of T cell acute lymphoblastic leukemia (T-ALL) patients, refractory and/or relapsing disease remains a foremost concern. In this context, a major contribution was provided by the introduction of the nucleoside analog nelarabine, approved for salvage treatment of T-ALL patients with refractory/relapsed disease. However, nelarabine could induce a life-threatening, dose-dependent neurotoxicity. To improve nelarabine efficacy, we have analyzed its molecular targets, testing selective inhibitors of such targets in combination with nelarabine.The effectiveness of nelarabine as single agent or in combination with PI3K, Bcl2, and MEK inhibitors was evaluated on human T-ALL cell lines and primary T-ALL refractory/relapsed lymphoblasts. The efficacy of signal modulators in terms of cytotoxicity, induction of apoptosis, and changes in gene and protein expression was assessed by flow cytometry, western blotting, and quantitative real-time PCR in T-ALL settings.Treatment with nelarabine as a single agent identified two groups of T-ALL cell lines, one sensitive and one resistant to the drug. Whereas sensitive T-ALL cells showed a significant increase of apoptosis and a strong down-modulation of PI3K signaling, resistant T-ALL cells showed a hyperactivation of AKT and MEK/ERK1/2 signaling pathways, not caused by differences in the expression of nelarabine transporters or metabolic activators. We then studied the combination of nelarabine with the PI3K inhibitors (both pan and dual γ/δ inhibitors), with the Bcl2 specific inhibitor ABT199, and with the MEK inhibitor trametinib on both T-ALL cell lines and patient samples at relapse, which displayed constitutive activation of PI3K signaling and resistance to nelarabine alone. The combination with the pan PI3K inhibitor ZSTK-474 was the most effective in inhibiting the growth of T-ALL cells and was synergistic in decreasing cell survival and inducing apoptosis in nelarabine-resistant T-ALL cells. The drug combination caused AKT dephosphorylation and a downregulation of Bcl2, while nelarabine alone induced an increase in p-AKT and Bcl2 signaling in the resistant T-ALL cells and relapsed patient samples.These findings indicate that nelarabine in combination with PI3K inhibitors may be a promising therapeutic strategy for the treatment of T-ALL relapsed patients.

    View details for DOI 10.1186/s13045-016-0344-4

    View details for Web of Science ID 000385896100001

    View details for PubMedID 27776559

    View details for PubMedCentralID PMC5075755

  • Human NK cells: From surface receptors to clinical applications. Immunology letters Moretta, L., Pietra, G., Vacca, P., Pende, D., Moretta, F., Bertaina, A., Mingari, M. C., Locatelli, F., Moretta, A. 2016; 178: 15-19


    Natural killer (NK) cells play a major role in innate defenses against pathogens, primarily viruses, and are also thought to be part of the immunosurveillance against tumors. They express an array of surface receptors that mediate NK cell function. The human leukocytes antigen (HLA) class I-specific inhibitory receptors allow NK cells to detect and kill cells that have lost or under-express HLA class I antigens, a typical feature of tumor or virally infected cells. However, NK cell activation and induction of cytolytic activity and cytokine production depends on another important checkpoint, namely the expression on target cells of ligands recognized by activating NK receptors. Despite their potent cytolytic activity, NK cells frequently fail to eliminate tumors. This is due to mechanisms of tumor escape, determined by the tumor cells themselves or by tumor-associated cells (i.e. the tumor microenvironment) via the release of soluble suppressive factors or the induction of inhibitory loops involving induction of regulatory T cells, M2-polarized macrophages and myeloid-derived suppressor cells. The most important clinical application involving NK cells is the cure of high-risk leukemias in the haplo-identical hematopoietic stem cell transplant (HSCT) setting. NK cells originated from hematopoietic stem cells (HSC) of HLA-haploidentical donors may express Killer Immunoglobulin-like receptors (KIRs) that are mismatched with the HLA class I alleles of the recipient. This allows NK cells to kill leukemia blasts residual after the conditioning regimen, while sparing normal cells (that do not express ligands for activating NK receptors). More recent approaches based on the specific removal of TCR α/β(+) T cells and of CD19(+) B cells, allow the infusion, together with CD34(+) HSC, of mature KIR(+) NK cells and of TCR γ/δ(+) T cells, both characterized by a potent anti-leukemia activity. This greatly reduces the time interval necessary to obtain alloreactive, KIR(+) NK cells derived from donor HSC. Another promising approach is based on the use of anti-KIR blocking monoclonal antibodies (mAbs), rendering alloreactive any KIR(+) NK cells.

    View details for DOI 10.1016/j.imlet.2016.05.007

    View details for PubMedID 27185471

  • Treatment Complications and Long-term Outcomes of Total Body Irradiation in Patients with Acute Lymphoblastic Leukemia: A Single Institute Experience ANTICANCER RESEARCH De Felice, F., Grapulin, L., Musio, D., Pomponi, J., Di Felice, C., Iori, A. P., Bertaina, A., Tombolini, V. 2016; 36 (9): 4859-4864


    The aim of this study was to evaluate treatment-related toxicity and clinical outcomes of total body irradiation (TBI) in patients with acute lymphoblastic leukemia (ALL).We performed a retrospective review of all patients with ALL who underwent TBI-based conditioning regimen at our Institution between 2000 and 2012.A total of 211 patients were included. The median follow-up was 40 months. The 5-year overall survival and disease-free survival were 64.7% and 62.8%, respectively. The 5-year overall survival rate for the 163 children was 67.6% (95% confidence interval=55-77%). Disease status at time of transplant did not improve disease-free survival. Gastrointestinal acute toxicity was the most common early side-effect (19.9%). Acute graft-versus-host disease was reported in 31 patients (14.7%). Main late toxicities were cataract induction (12.8%) and growth, gonadal and endocrine effects (36%).TBI-based conditioning regimen led to a high survival rate with remarkably low radiation-related toxicity, suggesting that TBI provides a feasible therapeutic option in patients with ALL.

    View details for DOI 10.21873/anticanres.11049

    View details for Web of Science ID 000384001800069

    View details for PubMedID 27630341

  • MicroRNA fingerprints in juvenile myelomonocytic leukemia (JMML) identified miR-150-5p as a tumor suppressor and potential target for treatment ONCOTARGET Leoncini, P. P., Bertaina, A., Papaioannou, D., Flotho, C., Masetti, R., Bresolin, S., Menna, G., Santoro, N., Zecca, M., Basso, G., Nigita, G., Veneziano, D., Pagotto, S., D'Ovidio, K., Rota, R., Dorrance, A., Croce, C. M., Niemeyer, C., Locatelli, F., Garzon, R. 2016; 7 (34): 55395-55408


    Juvenile myelomonocytic leukemia (JMML) is an aggressive leukemia of early childhood characterized by aberrant proliferation of myelomonocytic cells and hypersensitivity to GM-CSF stimulation. Mutually exclusive mutations in the RAS/ERK pathway genes such as PTPN11, NRAS, KRAS, CBL, or NF1 are found in ~90% of the cases. These mutations give rise to disease at least in part by activating STAT5 through phosphorylation and by promoting cell growth. MicroRNAs (miRs) are small non-coding RNAs that regulate gene expression, which are often deregulated in leukemia. However, little is known about their role in JMML. Here, we report distinctive miR expression signatures associated with the molecular subgroups of JMML. Among the downregulated miRs in JMML, miR-150-5p was found to target STAT5b, a gene which is often over-activated in JMML, and contributes to the characteristic aberrant signaling of this disorder. Moreover, loss of miR-150-5p and upregulation of STAT5b expression were also identified in a murine model of JMML. Ectopic overexpression of miR-150-5p in mononuclear cells from three JMML patients significantly decreased cell proliferation. Altogether, our data indicate that miR expression is deregulated in JMML and may play a role in the pathogenesis of this disorder by modulating key effectors of cytokine receptor pathways.

    View details for DOI 10.18632/oncotarget.10577

    View details for Web of Science ID 000385435000102

    View details for PubMedID 27447965

    View details for PubMedCentralID PMC5342425

  • Accuracy of Bedside Paediatric Early Warning System (BedsidePEWS) in a Pediatric Stem Cell Transplant Unit. Journal of pediatric oncology nursing Gawronski, O., Ciofi degli Atti, M. L., Di Ciommo, V., Cecchetti, C., Bertaina, A., Tiozzo, E., Raponi, M. 2016; 33 (4): 249-256


    Hospital mortality in children who undergo stem cell transplant (SCT) is high. Early warning scores aim at identifying deteriorating patients and at preventing adverse outcomes. The bedside pediatric early warning system (BedsidePEWS) is a pediatric early warning score based on 7 clinical indicators, ranging from 0 (all indicators within normal ranges for age) to 26. The aim of this case-control study was to assess the performance of BedsidePEWS in identifying clinical deterioration events among children admitted to an SCT unit. Cases were defined as clinical deterioration events; controls were all the other patients hospitalized on the same ward at the time of case occurrence. BedsidePEWS was retrospectively measured at 4-hour intervals in cases and controls 24 hours before an event (T4-T24). We studied 19 cases and 80 controls. The score significantly increased in cases from a median of 4 at T24 to a median of 14 at T4. The proportion of correctly classified cases and controls was >90% since T8. The area under the curve receiver operating characteristic was 0.9. BedsidePEWS is an accurate screening tool to predict clinical deterioration in SCT patients.

    View details for DOI 10.1177/1043454215600154

    View details for PubMedID 26497915

  • Posaconazole oral suspension primary prophylaxis in acute leukemia and allogeneic stem cell transplant patients: can it be used without measurement of plasma concentration? MEDICAL MYCOLOGY Girmenia, C., Annino, L., Mariotti, B., Fanci, R., Minotti, C., Spadea, A., Carotti, A., Piedimonte, M., Chierichini, A., Cerchiara, E., Caselli, D., Cupelli, L., Arcioni, F., Bertaina, A., Ribersani, M., Proia, A., Mengarelli, A., Perriello, V., Torelli, G. F., Di Gioia, M., Del Principe, M. I., Cassetta, M. I., Fallani, S., Novelli, A. 2016; 54 (5): 445-458


    Posaconazole oral suspension (PCZ-susp) can display a variable degree of inter and intra-individual absorption. However, there is no agreement on the need of plasma-posaconazole-concentration (PPC) monitoring as a routine practice in patients receiving PCZ-susp. In this prospective, multicenter study we evaluated the variability of PPCs in hematologic patients receiving PCZ-susp prophylaxis with the aim to define conditions at different risk of subtherapeutic PPCs. Overall, 103 acute leukemia (AL) patients submitted to intensive chemotherapy (115 courses) and 46 allogeneic stem cell transplant (allo-SCT) recipients (47 courses) receiving PCZ-susp prophylaxis were considered. The adequacy of PPC pattern after the steady state (≥day 7 of treatment) in courses with two or more PPC measurements was defined as follows: inadequate pattern: PPC < 0.5 mcg/ml at least once; borderline pattern: PPC always ≥0.5mcg/ml but < 0.7 mcg/ml at least once; adequate pattern: PPC always ≥0.7 mcg/ml. The PPC pattern was evaluable in 83 and 37 AL and allo-SCT patients, respectively. It was adequate, borderline and inadequate in 63.9%, 14.5%, and 21.7% of courses, respectively, in AL, and in 62.2%, 10.8%, and 27.0% of courses, respectively, in allo-SCT. In both groups, an inadequate PPC pattern was associated with the development of diarrhea. In absence of diarrhea, the probability of an inadequate PPC pattern was 11.9% in AL and 17.2% in allo-SCT patients. PCZ-susp might be used without stringent need of PPC monitoring in patients without diarrhea.

    View details for DOI 10.1093/mmy/myw001

    View details for Web of Science ID 000378865500001

    View details for PubMedID 26868905

  • Hematopoietic stem cell transplantation: Improving alloreactive Bw4 donor selection by genotyping codon 86 of KIR3DL1/S1. European journal of immunology Alicata, C., Pende, D., Meazza, R., Canevali, P., Loiacono, F., Bertaina, A., Locatelli, F., Nemat-Gorgani, N., Guethlein, L. A., Parham, P., Moretta, L., Moretta, A., Bottino, C., Norman, P. J., Falco, M. 2016; 46 (6): 1511-1517


    KIR3DL1 is a natural killer (NK) cell receptor that recognizes the Bw4 epitope of human leukocyte antigen (HLA) class I molecules. Following hematopoietic stem cell transplantation for patients lacking Bw4, KIR3DL1-expressing NK cells from Bw4-positive donors can be alloreactive and eliminate tumor cells. However, KIR3DL1 alleles having T instead of C at nucleotide 320 (encoding leucine 86 instead of serine 86) are not expressed on the cell surface. Thus, not all individuals testing positive for KIR3DL1 are optimal donors for Bw4-negative recipients. Therefore, we developed a method for genotyping codon 86, which was validated by its perfect correlation with NK cell phenotype for 100 donors of diverse KIR3DL1/S1 genotype. We typed 600 donors and found that ∼12.2% had the KIR3DL1 gene, but did not express cell-surface KIR3DL1. By contrast, high-expressing allotypes were identified when haplotypes from four families with duplicated KIR3DL1/S1 genes were characterized at high resolution. Identifying donors who have KIR3DL1 but lack cell-surface KIR3DL1 would refine donor selection. With this technique, the number of individuals identified who may not be optimal donors for Bw4-negative patients increases by threefold, when compared with standard methods. Taken together, we propose that allele typing of killer cell Ig-like receptor (KIR) polymorphisms should become a standard practice when selecting donors.

    View details for DOI 10.1002/eji.201546236

    View details for PubMedID 26990677

  • NK Cells and Other Innate Lymphoid Cells in Hematopoietic Stem Cell Transplantation FRONTIERS IN IMMUNOLOGY Vacca, P., Montaldo, E., Croxatto, D., Moretta, F., Bertaina, A., Vitale, C., Locatelli, F., Mingari, M. C., Moretta, L. 2016; 7


    Natural killer (NK) cells play a major role in the T-cell depleted haploidentical hematopoietic stem cell transplantation (haplo-HSCT) to cure high-risk leukemias. NK cells belong to the expanding family of innate lymphoid cells (ILCs). At variance with NK cells, the other ILC populations (ILC1/2/3) are non-cytolytic, while they secrete different patterns of cytokines. ILCs provide host defenses against viruses, bacteria, and parasites, drive lymphoid organogenesis, and contribute to tissue remodeling. In haplo-HSCT patients, the extensive T-cell depletion is required to prevent graft-versus-host disease (GvHD) but increases risks of developing a wide range of life-threatening infections. However, these patients may rely on innate defenses that are reconstituted more rapidly than the adaptive ones. In this context, ILCs may represent important players in the early phases following transplantation. They may contribute to tissue homeostasis/remodeling and lymphoid tissue reconstitution. While the reconstitution of NK cell repertoire and its role in haplo-HSCT have been largely investigated, little information is available on ILCs. Of note, CD34(+) cells isolated from different sources of HSC may differentiate in vitro toward various ILC subsets. Moreover, cytokines released from leukemia blasts (e.g., IL-1β) may alter the proportions of NK cells and ILC3, suggesting the possibility that leukemia may skew the ILC repertoire. Further studies are required to define the timing of ILC development and their potential protective role after HSCT.

    View details for DOI 10.3389/fimmu.2016.00188

    View details for Web of Science ID 000376060800001

    View details for PubMedID 27242795

    View details for PubMedCentralID PMC4870263

  • The generation of human innate lymphoid cells is influenced by the source of hematopoietic stem cells and by the use of G-CSF EUROPEAN JOURNAL OF IMMUNOLOGY Moretta, F., Petronelli, F., Lucarelli, B., Pitisci, A., Bertaina, A., Locatelli, F., Mingari, M. C., Moretta, L., Montaldo, E. 2016; 46 (5): 1271-1278


    NK cells play a central role in the haploidentical HSC transplantation (HSCT) to cure high-risk leukemias. Other innate lymphoid cells (ILCs) have been proposed to exert a protective role in graft-versus-host disease and could also contribute to anti-microbial defence and to lymphoid tissue remodeling. Thus, we investigated the ILC differentiation potential of HSCs isolated from BM, mobilized peripheral blood (PB), and umbilical cord blood (UCB). BM CD34(+) cells are enriched in lymphoid-committed precursors, while PB CD34(+) cells preferentially contain myeloid precursors. In vitro differentiation experiments revealed that the highest and the lowest CD56(+) CD161(+) ILC recovery was detected in UCB and PB HSC cultures, respectively. Among CD56(+) CD161(+) ILCs, the ratio between NK cells and ILC3s was similar for all HSC analyzed. ILC recovery in PB CD34(+) cultures was lower for G-CSF-mobilized HSCs (good mobilizers) than for G-CSF+plerixafor-mobilized HSC (poor mobilizers). Moreover, G-CSF inhibited in vitro ILC recovery and the degree of inhibition was proportional to the time of exposure to the cytokine. Thus, although all common sources of HSC for transplant differentiate towards ILCs, substantial differences exist among different sources and G-CSF may influence ILC recovery. These data offer new clues for a better understanding of the immune reconstitution after HSCT.

    View details for DOI 10.1002/eji.201546079

    View details for Web of Science ID 000380751700021

    View details for PubMedID 26840535

  • Human natural killer cells: news in the therapy of solid tumors and high-risk leukemias CANCER IMMUNOLOGY IMMUNOTHERAPY Pietra, G., Vitale, C., Pende, D., Bertaina, A., Moretta, F., Falco, M., Vacca, P., Montaldo, E., Cantoni, C., Mingari, M. C., Moretta, A., Locatelli, F., Moretta, L. 2016; 65 (4): 465-476


    It is well established that natural killer (NK) cells play an important role in the immunity against cancer, while the involvement of other recently identified, NK-related innate lymphoid cells is still poorly defined. In the haploidentical hematopoietic stem cell transplantation for the therapy of high-risk leukemias, NK cells have been shown to exert a key role in killing leukemic blasts residual after conditioning. While the clinical results in the cure of leukemias are excellent, the exploitation of NK cells in the therapy of solid tumors is still limited and unsatisfactory. In solid tumors, NK cell function may be inhibited via different mechanisms, occurring primarily at the tumor site. The cellular interactions in the tumor microenvironment involve tumor cells, stromal cells and resident or recruited leukocytes and may favor tumor evasion from the host's defenses. In this context, a number of cytokines, growth factors and enzymes synthesized by tumor cells, stromal cells, suppressive/regulatory myeloid and lymphoid cells may substantially impair the function of different tumor-reactive effector cells, including NK cells. The identification and characterization of such mechanisms may offer clues for the development of new immunotherapeutic strategies to restore effective anti-tumor responses. In order to harness NK cell-based immunotherapies, several approaches have been proposed, including reinforcement of NK cell cytotoxicity by means of specific cytokines, antibodies or drugs. These new tools may improve NK cell function and/or increase tumor susceptibility to NK-mediated killing. Hence, the integration of NK-based immunotherapies with conventional anti-tumor therapies may increase chances of successful cancer treatment.

    View details for DOI 10.1007/s00262-015-1744-y

    View details for Web of Science ID 000373952700010

    View details for PubMedID 26289090

  • Combined immunodeficiency due to JAK3 mutation in a child presenting with skin granuloma JOURNAL OF ALLERGY AND CLINICAL IMMUNOLOGY Scarselli, A., Di Cesare, S., Di Matteo, G., De Matteis, A., Ariganello, P., Romiti, M. L., Cascioli, S., De Vito, R., Bertaina, A., Locatelli, F., Gaspar, H. B., Aiuti, A., Rossi, P., Gilmour, K., Cancrini, C. 2016; 137 (3): 948-951

    View details for DOI 10.1016/j.jaci.2015.09.017

    View details for Web of Science ID 000371897500046

    View details for PubMedID 26545580

  • Analysis of memory-like natural killer cells in human cytomegalovirus-infected children undergoing aß+T and B cell-depleted hematopoietic stem cell transplantation for hematological malignancies. Haematologica Muccio, L., Bertaina, A., Falco, M., Pende, D., Meazza, R., Lopez-Botet, M., Moretta, L., Locatelli, F., Moretta, A., Della Chiesa, M. 2016; 101 (3): 371-381


    We analyzed the impact of human cytomegalovirus infection on the development of natural killer cells in 27 pediatric patients affected by hematological malignancies, who had received a HLA-haploidentical hematopoietic stem cell transplantation, depleted of both α/β+ T cells and B cells. In line with previous studies in adult recipients of umbilical cord blood transplantation, we found that human cytomegalovirus reactivation accelerated the emergence of mature natural killer cells. Thus, most children displayed a progressive expansion of a memory-like natural killer cell subset expressing NKG2C, a putative receptor for human cytomegalovirus, and CD57, a marker of terminal natural killer cell differentiation. NKG2C(+)CD57(+) natural killer cells were detectable by month 3 following hematopoietic stem cell transplantation and expanded until at least month 12. These cells were characterized by high killer Ig-like receptors (KIRs) and leukocyte inhibitory receptor 1 (LIR-1) and low Siglec-7, NKG2A and Interleukin-18Rα expression, killed tumor targets and responded to cells expressing HLA-E (a NKG2C ligand). In addition, they were poor Interferon-γ producers in response to Interleukin-12 and Interleukin-18. The impaired response to these cytokines, together with their highly differentiated profile, may reflect their skewing toward an adaptive condition specialized in controlling human cytomegalovirus. In conclusion, in pediatric patients receiving a type of allograft different from umbilical cord blood transplantation, human cytomegalovirus also induced memory-like natural killer cells, possibly contributing to controlling infections and reinforcing anti-leukemia effects.

    View details for DOI 10.3324/haematol.2015.134155

    View details for PubMedID 26659918

    View details for PubMedCentralID PMC4815729

  • Progressive increase in D-dimer levels during extracorporeal membrane oxygenation can predict membrane oxygenator failure in children given hematopoietic stem cell transplantation? JOURNAL OF CRITICAL CARE Di Nardo, M., Merli, P., Cecchetti, C., Pasotti, E., Bertaina, A., Locatelli, F. 2016; 31 (1): 262-263

    View details for DOI 10.1016/j.jcrc.2015.09.010

    View details for Web of Science ID 000366650500045

    View details for PubMedID 26476579

  • Synergistic cytotoxic effects of bortezomib and CK2 inhibitor CX-4945 in acute lymphoblastic leukemia: turning off the prosurvival ER chaperone BIP/Grp78 and turning on the pro-apoptotic NF-kappa B ONCOTARGET Buontempo, F., Orsini, E., Lonetti, A., Cappellini, A., Chiarini, F., Evangelisti, C., Evangelisti, C., Melchionda, F., Pession, A., Bertaina, A., Locatelli, F., Bertacchini, J., Neri, L. M., McCubrey, J. A., Martelli, A. M. 2016; 7 (2): 1323-1340


    The proteasome inhibitor bortezomib is a new targeted treatment option for refractory or relapsed acute lymphoblastic leukemia (ALL) patients. However, a limited efficacy of bortezomib alone has been reported. A terminal pro-apoptotic endoplasmic reticulum (ER) stress/unfolded protein response (UPR) is one of the several mechanisms of bortezomib-induced apoptosis. Recently, it has been documented that UPR disruption could be considered a selective anti-leukemia therapy. CX-4945, a potent casein kinase (CK) 2 inhibitor, has been found to induce apoptotic cell death in T-ALL preclinical models, via perturbation of ER/UPR pathway. In this study, we analyzed in T- and B-ALL preclinical settings, the molecular mechanisms of synergistic apoptotic effects observed after bortezomib/CX-4945 combined treatment. We demonstrated that, adding CX-4945 after bortezomib treatment, prevented leukemic cells from engaging a functional UPR in order to buffer the bortezomib-mediated proteotoxic stress in ER lumen. We documented that the combined treatment decreased pro-survival ER chaperon BIP/Grp78 expression, via reduction of chaperoning activity of Hsp90. Bortezomib/CX-4945 treatment inhibited NF-κB signaling in T-ALL cell lines and primary cells from T-ALL patients, but, intriguingly, in B-ALL cells the drug combination activated NF-κB p65 pro-apoptotic functions. In fact in B-cells, the combined treatment induced p65-HDAC1 association with consequent repression of the anti-apoptotic target genes, Bcl-xL and XIAP. Exposure to NEMO (IKKγ)-binding domain inhibitor peptide reduced the cytotoxic effects of bortezomib/CX-4945 treatment. Overall, our findings demonstrated that CK2 inhibition could be useful in combination with bortezomib as a novel therapeutic strategy in both T- and B-ALL.

    View details for Web of Science ID 000369951100020

    View details for PubMedID 26593250

    View details for PubMedCentralID PMC4811463

  • Haploidentical Haematopoietic Stem Cell Transplantation: Role of NK Cells and Effect of Cytomegalovirus Infections NATURAL KILLER CELLS Della Chiesa, M., Moretta, L., Muccio, L., Bertaina, A., Moretta, F., Locatelli, F., Moretta, A. 2016; 395: 209-224


    Natural killer cells play an important role in the immune responses against cancer and viral infections. In addition, NK cells have been shown to exert a key role in haploidentical hematopoietic stem cell (HSC) transplantation for the therapy of high-risk leukemias. The anti-leukemia effect is mostly related to the presence of "alloreactive" NK cells, i.e., mature KIR(+) NK cells that express inhibitory KIR mismatched with HLA class I (KIR-L) of the patient. In addition, an important role is played by certain activating KIR (primarily, but not only, KIR2DS1) upon interaction with their HLA class I ligand (C2 alleles). In general, the presence of activating KIR correlates with a better prognosis. Beside the infusion of "pure" CD34(+) cells, a novel protocol has been recently developed in which depletion of αβ T cells and CD19(+) B cells makes it possible to infuse into the patient, together with donor CD34(+) HSCs, important effector cells including mature PB NK cells and γδ T cells. Recent studies revealed that cytomegalovirus (CMV) infection/reactivation may induce rapid NK cell maturation and greatly influence the NK receptor repertoire. The remarkable expansion of a subset expressing the activating receptor NKG2C, together with a more efficient virus-specific effector response after rechallenge with CMV (i.e., antigen specificity), and the longevity of the expanded population are all features consistent with an adaptive type of response and support the notion of a memory-like activity of NK cells.

    View details for DOI 10.1007/82_2015_450

    View details for Web of Science ID 000385417700010

    View details for PubMedID 26160014

  • MicroRNA-101 is repressed by EZH2 and its restoration inhibits tumorigenic features in embryonal rhabdomyosarcoma CLINICAL EPIGENETICS Vella, S., Pomella, S., Leoncini, P. P., Colletti, M., Conti, B., Marquez, V. E., Strillacci, A., Roma, J., Gallego, S., Milano, G. M., Capogrossi, M. C., Bertaina, A., Ciarapica, R., Rota, R. 2015; 7


    Rhabdomyosarcoma (RMS) is a pediatric soft tissue sarcoma arising from myogenic precursors that have lost their capability to differentiate into skeletal muscle. The polycomb-group protein EZH2 is a Lys27 histone H3 methyltransferase that regulates the balance between cell proliferation and differentiation by epigenetically silencing muscle-specific genes. EZH2 is often over-expressed in several human cancers acting as an oncogene. We previously reported that EZH2 inhibition induces cell cycle arrest followed by myogenic differentiation of RMS cells of the embryonal subtype (eRMS). MiR-101 is a microRNA involved in a negative feedback circuit with EZH2 in different normal and tumor tissues. To that, miR-101 can behave as a tumor suppressor in several cancers by repressing EZH2 expression. We, therefore, evaluated whether miR-101 is de-regulated in eRMS and investigated its interplaying with EZH2 as well as its role in the in vitro tumorigenic potential of these tumor cells.Herein, we report that miR-101 is down-regulated in eRMS patients and in tumor cell lines compared to their controls showing an inverse pattern of expression with EZH2. We also show that miR-101 is up-regulated in eRMS cells following both genetic and pharmacological inhibition of EZH2. In turn, miR-101 forced expression reduces EZH2 levels as well as restrains the migratory potential of eRMS cells and impairs their clonogenic and anchorage-independent growth capabilities. Finally, EZH2 recruitment to regulatory region of miR-101-2 gene decreases in EZH2-silenced eRMS cells. This phenomenon is associated to reduced H3K27me3 levels at the same regulatory locus, indicating that EZH2 directly targets miR-101 for repression in eRMS cells.Altogether, our data show that, in human eRMS, miR-101 is involved in a negative feedback loop with EZH2, whose targeting has been previously shown to halt eRMS tumorigenicity. They also demonstrate that the re-induction of miR-101 hampers the tumor features of eRMS cells. In this scenario, epigenetic dysregulations confirm their crucial role in the pathogenesis of this soft tissue sarcoma.

    View details for DOI 10.1186/s13148-015-0107-z

    View details for Web of Science ID 000359104600001

    View details for PubMedID 26251675

    View details for PubMedCentralID PMC4527101

  • ?d T-cell reconstitution after HLA-haploidentical hematopoietic transplantation depleted of TCR-aß+/CD19+ lymphocytes. Blood Airoldi, I., Bertaina, A., Prigione, I., Zorzoli, A., Pagliara, D., Cocco, C., Meazza, R., Loiacono, F., Lucarelli, B., Bernardo, M. E., Barbarito, G., Pende, D., Moretta, A., Pistoia, V., Moretta, L., Locatelli, F. 2015; 125 (15): 2349-2358


    We prospectively assessed functional and phenotypic characteristics of γδ T lymphocytes up to 7 months after HLA-haploidentical hematopoietic stem cell transplantation (haplo-HSCT) depleted of αβ(+) T cells and CD19(+) B cells in 27 children with either malignant or nonmalignant disorders. We demonstrate that (1) γδ T cells are the predominant T-cell population in patients during the first weeks after transplantation, being mainly, albeit not only, derived from cells infused with the graft and expanding in vivo; (2) central-memory cells predominated very early posttransplantation for both Vδ1 and Vδ2 subsets; (3) Vδ1 cells are specifically expanded in patients experiencing cytomegalovirus reactivation and are more cytotoxic compared with those of children who did not experience reactivation; (4) these subsets display a cytotoxic phenotype and degranulate when challenged with primary acute myeloid and lymphoid leukemia blasts; and (5) Vδ2 cells are expanded in vitro after exposure to zoledronic acid (ZOL) and efficiently lyse primary lymphoid and myeloid blasts. This is the first detailed characterization of γδ T cells emerging in peripheral blood of children after CD19(+) B-cell and αβ(+) T-cell-depleted haplo-HSCT. Our results can be instrumental to the development of clinical trials using ZOL for improving γδ T-cell killing capacity against leukemia cells. This trial was registered at as #NCT01810120.

    View details for DOI 10.1182/blood-2014-09-599423

    View details for PubMedID 25612623

  • Multifunctional human CD56 low CD16 low natural killer cells are the prominent subset in bone marrow of both healthy pediatric donors and leukemic patients. Haematologica Stabile, H., Nisti, P., Morrone, S., Pagliara, D., Bertaina, A., Locatelli, F., Santoni, A., Gismondi, A. 2015; 100 (4): 489-498


    We phenotypically and functionally characterized a distinct CD56(low) natural killer cell subset based on CD16 expression levels in bone marrow and peripheral blood of healthy children and pediatric patients with acute lymphoblastic leukemia. Our findings demonstrate for the first time that CD56(low)CD16(low) natural killer cells are more abundant in bone marrow than in peripheral blood and that their frequency is further increased in children with acute lymphoblastic leukemia. Bone marrow and peripheral blood CD56(low)CD16(low) natural killer cells compared with CD56(low)CD16(high) natural killer cells express lower levels of killer inhibitory receptors, higher levels of CD27, CD127, CD122, CD25, but undetectable levels of CD57, suggesting that they have a higher proliferative and differentiation potential. Moreover, CD56(low)CD16(low) natural killer cells display higher levels of CXCR4 and undetectable levels of CX3CR1 and can be consistently and rapidly mobilized in peripheral blood in response to CXCR4 antagonist. Unlike CD56(low)CD16(high), both bone marrow and peripheral blood CD56(low)CD16(low) natural killer cells release IFNγ following cytokine stimulation, and represent the major cytotoxic natural killer cell population against K562 or acute lymphoblastic leukemia target cells. All these data suggest that CD56(low)CD16(low) natural killer cells are multifunctional cells, and that the presence of hematologic malignancies affects their frequency and functional ability at both tumor site and in the periphery.

    View details for DOI 10.3324/haematol.2014.116053

    View details for PubMedID 25596273

    View details for PubMedCentralID PMC4380722

  • Infections by carbapenem-resistant Klebsiella pneumoniae in SCT recipients: a nationwide retrospective survey from Italy BONE MARROW TRANSPLANTATION Girmenia, C., Rossolini, G. M., Piciocchi, A., Bertaina, A., Pisapia, G., Pastore, D., Sica, S., Severino, A., Cudillo, L., Ciceri, F., Scime, R., Lombardini, L., Viscoli, C., Rambaldi, A. 2015; 50 (2): 282-288


    Infections by carbapenem-resistant Klebsiella pneumoniae (CRKp) represent a challenging problem after SCT. A retrospective survey (January 2010 to July 2013) involving 52 Italian centers was performed to assess the epidemiology and the prognostic factors of CRKp infections in auto- and allo-SCT. Cases of CRKp infection were reported in 53.4% of centers. CRKp infections were documented in 25 auto-SCTs and 87 allo-SCTs, with an incidence of 0.4% (from 0.1% in 2010 to 0.7% in 2013) and 2% (from 0.4% in 2010 to 2.9% in 2013), respectively. A CRKp colonization documented before or after transplant was followed by an infection in 25.8% of auto-SCT and 39.2% of allo-SCT patients. The infection-related mortality rates were 16% and 64.4%, respectively. A pre-transplant CRKp infection (hazard ratio (HR) 0.33, 95% confidence intervals (CIs) 0.15-0.74; P=0.007) and a not CRKp-targeted first-line treatment (HR 2.67, 95% CI 1.43-4.99; P=0.002) were independent factors associated with an increased mortality in allo-SCT patients who developed a CRKp infection. Our study shows challenging findings of CRKp infections in SCT patients in Italy particularly after allo-SCT. The detection of carriers and the definition of early therapeutic strategies represent critical aspects of the management of CRKp infections after SCT.

    View details for DOI 10.1038/bmt.2014.231

    View details for Web of Science ID 000349402900021

    View details for PubMedID 25310302

  • Outcome of children with high-risk acute myeloid leukemia given autologous or allogeneic hematopoietic cell transplantation in the aieop AML-2002/01 study BONE MARROW TRANSPLANTATION Locatelli, F., Masetti, R., Rondelli, R., Zecca, M., Fagioli, F., Rovelli, A., Messina, C., Lanino, E., Bertaina, A., Favre, C., Giorgiani, G., Ripaldi, M., Ziino, O., Palumbo, G., Pillon, M., Pession, A., Rutella, S., Prete, A. 2015; 50 (2): 181-188


    We analyzed the outcome of 243 children with high-risk (HR) AML in first CR1 enrolled in the AIEOP-2002/01 protocol, who were given either allogeneic (ALLO; n=141) or autologous (AUTO; n=102) hematopoietic SCT (HSCT), depending on the availability of a HLA-compatible sibling. Infants, patients with AML-M7, or complex karyotype or those with FLT3-ITD, were eligible to be transplanted also from alternative donors. All patients received a myeloablative regimen combining busulfan, cyclophosphamide and melphalan; [corrected] AUTO-HSCT patients received BM cells in most cases, while in children given ALLO-HSCT stem cell source was BM in 96, peripheral blood in 19 and cord blood in 26. With a median follow-up of 57 months (range 12-130), the probability of disease-free survival (DFS) was 73% and 63% in patients given either ALLO- or AUTO-HSCT, respectively (P=NS). Although the cumulative incidence (CI) of relapse was lower in ALLO- than in AUTO-HSCT recipients (17% vs 28%, respectively; P=0.043), the CI of TRM was 7% in both groups. Patients transplanted with unrelated donor cord blood had a remarkable 92.3% 8-year DFS probability. Altogether, these data confirm that HSCT is a suitable option for preventing leukemia recurrence in HR children with CR1 AML.

    View details for DOI 10.1038/bmt.2014.246

    View details for Web of Science ID 000349402900005

    View details for PubMedID 25387094

  • KIR B haplotype donors confer a reduced risk for relapse after haploidentical transplantation in children with ALL BLOOD Oevermann, L., Michaelis, S. U., Mezger, M., Lang, P., Toporski, J., Bertaina, A., Zecca, M., Moretta, L., Locatelli, F., Handgretinger, R. 2014; 124 (17): 2744-2747


    We analyzed the influence of donor killer-cell immunoglobulin-like receptor (KIR) gene haplotypes on the risk for relapse and the probability of event-free survival (EFS) in children with acute lymphoblastic leukemia who received human leukocyte antigen-haploidentical transplantation of ex vivo T-cell-depleted peripheral blood stem cells. The KIR gene haplotype was evaluated in 85 donors, and the KIR B content score was determined in the 63 KIR haplotype B donors. Patients transplanted from a KIR haplotype B donor had a significantly better EFS than those transplanted from a KIR haplotype A donor (50.6% vs 29.5%, respectively; P = .033). Moreover, a high donor KIR B-content score was associated with a significantly reduced risk for relapse (Log-rank test for trend, P = .026). These data indicate that KIR genotyping should be included in the donor selection algorithm for haploidentical transplantation in children with acute lymphoblastic leukemia with the aim of choosing, whenever possible, a KIR haplotype B donor with a high KIR B-content score.

    View details for DOI 10.1182/blood-2014-03-565069

    View details for Web of Science ID 000347457500021

    View details for PubMedID 25115891

    View details for PubMedCentralID PMC4208288

  • Extracorporeal membrane oxygenation as a bridge to allogeneic T-cell depleted hematopoietic stem cell transplantation in infants with severe combined immune deficiency: is it feasible? INTENSIVE CARE MEDICINE Di Nardo, M., Locatelli, F., Di Florio, F., Cecchetti, C., Amodeo, A., Rutella, S., Bertaina, A. 2014; 40 (10): 1600-1601

    View details for DOI 10.1007/s00134-014-3432-3

    View details for Web of Science ID 000342433000034

    View details for PubMedID 25112503

  • Assessment of the effect of sphingosine kinase inhibitors on apoptosis, unfolded protein response and autophagy of T-cell acute lymphoblastic leukemia cells; indications for novel therapeutics ONCOTARGET Evangelisti, C., Evangelisti, C., Teti, G., Chiarini, F., Falconi, M., Melchionda, F., Pession, A., Bertaina, A., Locatelli, F., McCubrey, J. A., Beak, D. J., Bittman, R., Pyne, S., Pyne, N. J., Martelli, A. M. 2014; 5 (17): 7886-7901


    Sphingosine 1-phosphate (S1P) is a bioactive lipid that is formed by the phosphorylation of sphingosine and catalysed by sphingosine kinase 1 (SK1) or sphingosine kinase 2 (SK2). Sphingosine kinases play a fundamental role in many signaling pathways associated with cancer, suggesting that proteins belonging to this signaling network represent potential therapeutic targets. Over the last years, many improvements have been made in the treatment of T-cell acute lymphoblastic leukemia (T-ALL); however, novel and less toxic therapies are still needed, especially for relapsing and chemo-resistant patients. Here, we analyzed the therapeutic potential of SKi and ROMe, a sphingosine kinase 1 and 2 inhibitor and SK2-selective inhibitor, respectively. While SKi induced apoptosis, ROMe initiated an autophagic cell death in our in vitro cell models. SKi treatment induced an increase in SK1 protein levels in Molt-4 cells, whereas it activated the endoplasmic reticulum (ER) stress/unfolded protein response (UPR) pathway in Jurkat and CEM-R cells as protective mechanisms in a sub-population of T-ALL cells. Interestingly, we observed a synergistic effect of SKi with the classical chemotherapeutic drug vincristine. In addition, we reported that SKi affected signaling cascades implicated in survival, proliferation and stress response of cells. These findings indicate that SK1 or SK2 represent potential targets for treating T-ALL.

    View details for Web of Science ID 000348029800052

    View details for PubMedID 25226616

    View details for PubMedCentralID PMC4202168

  • An outbreak of extremely drug-resistant Pseudomonas aeruginosa in a tertiary care pediatric hospital in Italy BMC INFECTIOUS DISEASES degli Atti, M. C., Bernaschi, P., Carletti, M., Luzzi, I., Garcia-Fernandez, A., Bertaina, A., Sisto, A., Locatelli, F., Raponi, M. 2014; 14


    Extensively drug-resistant Pseudomonas aeruginosa (XDR-PA) isolates are susceptible to only one or two classes of antibiotics. In 2011-2012, we investigated an outbreak of XDR-PA affecting children with onco-hematological diseases.Outbreak investigation included ascertainment of cases, tracing of intestinal carriers and environmental surveillance. Contact precautions were adopted for patients with infection or colonization. Isolates were tested for antimicrobial susceptibility; phenotypic confirmation of carbapenemase production was performed, and carbapenemase genes were tested by multiplex polymerase-chain-reaction (PCR). Genotypes were determined by pulsed-field gel electrophoresis (PFGE).XDR-PA was isolated from 27 patients; 12 had bacteremia, 6 other infections and 9 were colonized. Severe neutropenia was significantly associated with bacteremia. Bloodstream-infection mortality rate was 67%. All isolates were resistant to carbapenems, cephalosporins and penicillins + β-lactamase inhibitors. Isolates were susceptible only to colistin in 22 patients, to colistin and amikacin in 4, and to ciprofloxacin and colistin in 1. PFGE results identified 6 subtypes of a single genotype, associated with clusters of cases, and 4 sporadic genotypes. Two sporadic isolates were metallo-β-lactamase producers, negative to PCR. All other isolates were metallo-β-lactamase producers due to the presence of a VIM carbapenemase. Incidence of XDR-PA infections decreased from 0.72 cases/1,000 inpatient-days in March 2011-March 2012, to 0.34/1,000 in April-December 2012, after implementation of active finding of intestinal carriers on all onco-hematological inpatients.Control measures targeting intestinal carriers are crucial in limiting in-hospital transmission of XDR-PA polyclonal strains, protecting more vulnerable patients, such as severely neutropenic children, from developing clinical infections.

    View details for DOI 10.1186/1471-2334-14-494

    View details for Web of Science ID 000341605000001

    View details for PubMedID 25209325

    View details for PubMedCentralID PMC4167521

  • Mobilization of healthy donors with plerixafor affects the cellular composition of T-cell receptor (TCR)-alpha beta/CD19-depleted haploidentical stem cell grafts JOURNAL OF TRANSLATIONAL MEDICINE Rutella, S., Filippini, P., Bertaina, V., Li Pira, G., Altomare, L., Ceccarelli, S., Brescia, L. P., Lucarelli, B., Girolami, E., Conflitti, G., Cefalo, M. G., Bertaina, A., Corsetti, T., Moretta, L., Locatelli, F. 2014; 12


    HLA-haploidentical hematopoietic stem cell transplantation (HSCT) is suitable for patients lacking related or unrelated HLA-matched donors. Herein, we investigated whether plerixafor (MZ), as an adjunct to G-CSF, facilitated the collection of mega-doses of hematopoietic stem cells (HSC) for TCR-αβ/CD19-depleted haploidentical HSCT, and how this agent affects the cellular graft composition.Ninety healthy donors were evaluated. Single-dose MZ was given to 30 'poor mobilizers' (PM) failing to attain ≥40 CD34+ HSCs/μL after 4 daily G-CSF doses and/or with predicted apheresis yields ≤12.0x106 CD34+ cells/kg recipient's body weight.MZ significantly increased CD34+ counts in PM. Naïve/memory T and B cells, as well as natural killer (NK) cells, myeloid/plasmacytoid dendritic cells (DCs), were unchanged compared with baseline. MZ did not further promote the G-CSF-induced mobilization of CD16+ monocytes and the down-regulation of IFN-γ production by T cells. HSC grafts harvested after G-CSF + MZ were enriched in myeloid and plasmacytoid DCs, but contained low numbers of pro-inflammatory 6-sulfo-LacNAc+ (Slan)-DCs. Finally, children transplanted with G-CSF + MZ-mobilized grafts received greater numbers of monocytes, myeloid and plasmacytoid DCs, but lower numbers of NK cells, NK-like T cells and Slan-DCs.MZ facilitates the collection of mega-doses of CD34+ HSCs for haploidentical HSCT, while affecting graft composition.

    View details for DOI 10.1186/s12967-014-0240-z

    View details for Web of Science ID 000341663000001

    View details for PubMedID 25179788

    View details for PubMedCentralID PMC4158047

  • HLA-haploidentical stem cell transplantation after removal of alpha beta(+) T and B cells in children with nonmalignant disorders BLOOD Bertaina, A., Merli, P., Rutella, S., Pagliara, D., Bernardo, M. E., Masetti, R., Pende, D., Falco, M., Handgretinger, R., Moretta, F., Lucarelli, B., Brescia, L. P., Li Pira, G., Testi, M., Cancrini, C., Kabbara, N., Carsetti, R., Finocchi, A., Moretta, A., Moretta, L., Locatelli, F. 2014; 124 (5): 822-826


    Twenty-three children with nonmalignant disorders received HLA-haploidentical hematopoietic stem cell transplantation (haplo-HSCT) after ex vivo elimination of αβ(+) T cells and CD19(+) B cells. The median number of CD34(+), αβ(+)CD3(+), and B cells infused was 16.8 × 10(6), 40 × 10(3), and 40 × 10(3) cells/kg, respectively. No patient received any posttransplantation pharmacologic prophylaxis for graft-versus-host disease (GVHD). All but 4 patients engrafted, these latter being rescued by a second allograft. Three patients experienced skin-only grade 1 to 2 acute GVHD. No patient developed visceral acute or chronic GVHD. Cumulative incidence of transplantation-related mortality was 9.3%. With a median follow-up of 18 months, 21 of 23 children are alive and disease-free, the 2-year probability of disease-free survival being 91.1%. Recovery of γδ(+) T cells was prompt, but αβ(+) T cells progressively ensued over time. Our data suggest that this novel graft manipulation strategy is safe and effective for haplo-HSCT. This trial was registered at as #NCT01810120.

    View details for DOI 10.1182/blood-2014-03-563817

    View details for Web of Science ID 000342619900026

    View details for PubMedID 24869942

  • Therapeutic targeting of Polo-like kinase-1 and Aurora kinases in T-cell acute lymphoblastic leukemia CELL CYCLE Sparta, A. M., Bressanin, D., Chiarini, F., Lonetti, A., Cappellini, A., Evangelisti, C., Evangelisti, C., Melchionda, F., Pession, A., Bertaina, A., Locatelli, F., McCubrey, J. A., Martelli, A. M. 2014; 13 (14): 2237-2247


    Polo-like kinases (PLKs) and Aurora kinases (AKs) act as key cell cycle regulators in healthy human cells. In cancer, these protein kinases are often overexpressed and dysregulated, thus contributing to uncontrolled cell proliferation and growth. T-cell acute lymphoblastic leukemia (T-ALL) is a heterogeneous malignancy arising in the thymus from T-cell progenitors. Primary chemoresistant and relapsed T-ALL patients have yet a poor outcome, therefore novel therapies, targeting signaling pathways important for leukemic cell proliferation, are required. Here, we demonstrate the potential therapeutic effects of BI6727, MK-5108, and GSK1070916, three selective inhibitors of PLK1, AK-A, and AK-B/C, respectively, in a panel of T-ALL cell lines and primary cells from T-ALL patients. The drugs were both cytostatic and cytotoxic to T-ALL cells by inducing G2/M-phase arrest and apoptosis. The drugs retained part of their pro-apoptotic activity in the presence of MS-5 bone marrow stromal cells. Moreover, we document for the first time that BI6727 perturbed both the PI3K/Akt/mTORC2 and the MEK/ERK/mTORC1 signaling pathways, and that a combination of BI6727 with specific inhibitors of the aforementioned pathways (MK-2206, CCI-779) displayed significantly synergistic cytotoxic effects. Taken together, our findings indicate that PLK1 and AK inhibitors display the potential for being employed in innovative therapeutic strategies for improving T-ALL patient outcome.

    View details for DOI 10.4161/cc.29267

    View details for Web of Science ID 000340272100015

    View details for PubMedID 24874015

    View details for PubMedCentralID PMC4111679

  • Recognition of adult and pediatric acute lymphoblastic leukemia blasts by natural killer cells HAEMATOLOGICA Torelli, G. F., Peragine, N., Raponi, S., Pagliara, D., De Propris, M. S., Vitale, A., Bertaina, A., Barberi, W., Moretta, L., Basso, G., Santoni, A., Guarini, A., Locatelli, F., Foa, R. 2014; 99 (7): 1248-1254


    In this study, we aimed to investigate the pathways of recognition of acute lymphoblastic leukemia blasts by natural killer cells and to verify whether differences in natural killer cell activating receptor ligand expression among groups defined by age of patients, or presence of cytogenetic/molecular aberrations correlate with the susceptibility to recognition and killing. We analyzed 103 newly diagnosed acute lymphoblastic leukemia patients: 46 adults and 57 children. Pediatric blasts showed a significantly higher expression of Nec-2 (P=0.03), ULBP-1 (P=0.01) and ULBP-3 (P=0.04) compared to adult cells. The differential expression of these ligands between adults and children was confined to B-lineage acute lymphoblastic leukemia with no known molecular alterations. Within molecularly defined subgroups of patients, a high surface expression of NKG2D and DNAM1 ligands was found on BCR-ABL(+) blasts, regardless of patient age. Accordingly, BCR-ABL(+) blasts proved to be significantly more susceptible to natural killer-dependent lysis than B-lineage blasts without molecular aberrations (P=0.03). Cytotoxic tests performed in the presence of neutralizing antibodies indicated a pathway of acute lymphoblastic leukemia cell recognition in the setting of the Nec-2/DNAM-1 interaction. These data provide a biological explanation of the different roles played by alloreactive natural killer cells in pediatric versus adult acute lymphoblastic leukemia and suggest that new natural killer-based strategies targeting specific subgroups of patients, particularly those BCR-ABL(+), are worth pursuing further.

    View details for DOI 10.3324/haematol.2013.101931

    View details for Web of Science ID 000342833500030

    View details for PubMedID 24658822

    View details for PubMedCentralID PMC4077088

  • Activity of the pan-class I phosphoinositide 3-kinase inhibitor NVP-BKM120 in T-cell acute lymphoblastic leukemia LEUKEMIA Lonetti, A., Antunes, I. L., Chiarini, F., Orsini, E., Buontempo, F., Ricci, F., Tazzari, P. L., Pagliaro, P., Melchionda, F., Pession, A., Bertaina, A., Locatelli, F., McCubrey, J. A., Barata, J. T., Martelli, A. M. 2014; 28 (6): 1196-1206


    Constitutively active phosphoinositide 3-kinase (PI3K) signaling is a common feature of T-cell acute lymphoblastic leukemia (T-ALL), where it upregulates cell proliferation, survival and drug resistance. These observations lend compelling weight to the application of PI3K inhibitors in the therapy of T-ALL. Here, we have analyzed the therapeutic potential of the pan-PI3K inhibitor NVP-BKM120 (BKM120), an orally bioavailable 2,6-dimorpholino pyrimidine derivative, which has entered clinical trials for solid tumors, on both T-ALL cell lines and patient samples. BKM120 treatment resulted in G2/M phase cell cycle arrest and apoptosis, being cytotoxic to a panel of T-ALL cell lines and patient T lymphoblasts, and promoting a dose- and time-dependent dephosphorylation of Akt and S6RP. BKM120 maintained its pro-apoptotic activity against Jurkat cells even when cocultured with MS-5 stromal cells, which mimic the bone marrow microenvironment. Remarkably, BKM120 synergized with chemotherapeutic agents currently used for treating T-ALL patients. Moreover, in vivo administration of BKM120 to a subcutaneous xenotransplant model of human T-ALL significantly delayed tumor growth, thus prolonging survival time. Taken together, our findings indicate that BKM120, either alone or in combination with chemotherapeutic drugs, may be an efficient treatment for T-ALLs that have aberrant upregulation of the PI3K signaling pathway.

    View details for DOI 10.1038/leu.2013.369

    View details for Web of Science ID 000337230300003

    View details for PubMedID 24310736

  • Extracorporeal membrane oxygenation in pediatric recipients of hematopoietic stem cell transplantation: an updated analysis of the Extracorporeal Life Support Organization experience INTENSIVE CARE MEDICINE Di Nardo, M., Locatelli, F., Palmer, K., Amodeo, A., Lorusso, R., Belliato, M., Cecchetti, C., Perrotta, D., Picardo, S., Bertaina, A., Rutella, S., Rycus, P., Di Ciommo, V., Holzgraefe, B. 2014; 40 (5): 754-756

    View details for DOI 10.1007/s00134-014-3240-9

    View details for Web of Science ID 000335662400020

    View details for PubMedID 24556913

  • HLA-Haploidentical T Cell-Depleted Allogeneic Hematopoietic Stem Cell Transplantation in Children with Fanconi Anemia BIOLOGY OF BLOOD AND MARROW TRANSPLANTATION Zecca, M., Strocchio, L., Pagliara, D., Comoli, P., Bertaina, A., Giorgiani, G., Perotti, C., Corbella, F., Brescia, L., Locatelli, F. 2014; 20 (4): 571-576


    We report the outcome of 12 consecutive pediatric patients with Fanconi anemia (FA) who had neither an HLA-identical sibling nor an HLA-matched unrelated donor and who were given T cell-depleted, CD34(+) positively selected cells from a haploidentical related donor after a reduced-intensity, fludarabine-based conditioning regimen. Engraftment was achieved in 9 of 12 patients (75%), and the cumulative incidence of graft rejection was 17% (95% confidence interval [CI], 5% to 59%). Cumulative incidences of grades II to IV acute and chronic graft-versus-host disease were 17% (95% CI, 5% to 59%) and 35% (95% CI, 14% to 89%), respectively. The conditioning regimen was well tolerated, with no fatal regimen-related toxicity and 3 cases of grade III regimen-related toxicity. The cumulative incidence of transplant-related mortality was 17% (95% CI, 5% to 59%). The 5-year overall survival, event-free survival, and disease-free survival were 83% (95% CI, 62% to 100%), 67% (95% CI, 40% to 93%), and 83% (95% CI, 62% to 100%), respectively. These data demonstrate that a fludarabine-based conditioning regimen, followed by infusion of high doses of T cell-depleted stem cells, is able to ensure engraftment with good overall survival and disease-free survival, confirming the feasibility of haploidentical hematopoietic stem cell transplantation in FA. To the best of our knowledge, this is the largest series of hematopoietic stem cell transplantation from a haploidentical related donor in FA patients reported to date.

    View details for DOI 10.1016/j.bbmt.2014.01.015

    View details for Web of Science ID 000333322400020

    View details for PubMedID 24462983

  • Familial Hemophagocytic Lymphohistiocytosis Type 3 Diagnosed at School Age: A Case Report JOURNAL OF PEDIATRIC HEMATOLOGY ONCOLOGY Manno, E. C., Salfa, I., Palma, P., Bertaina, A., Lombardi, A., Moretta, F., Coniglio, M. L., Sieni, E., Arico, M., Finocchi, A. 2014; 36 (2): E128-E130


    Familial hemophagocytic lymphohistiocytosis (HLH) is a rare, life-threatening condition characterized by immune hyperactivation and clinical signs of extreme inflammation. We describe a 7-year-old male who presented with fever resistant to antibiotic therapy, pancytopenia, splenomegaly, hypertriglyceridemia, and hyperferritinemia. Bone marrow aspirate showed hemophagocytosis. Epstein-Barr virus genome was positive in blood. Functional screening showed reduced capacity of cytotoxic degranulation. Mutation analysis of the FHL-related genes revealed compound heterozygous for UNC13D mutations: c. 753+1G>T, and the novel c.544C>T (p.P182S). Patients with a clinical presentation of HLH, even if older than typically seen, should be screened for familial HLH by mutation analysis.

    View details for DOI 10.1097/MPH.0b013e318292bc7c

    View details for Web of Science ID 000332087400026

    View details for PubMedID 23669735

  • Cytotoxic activity of the casein kinase 2 inhibitor CX-4945 against T-cell acute lymphoblastic leukemia: targeting the unfolded protein response signaling LEUKEMIA Buontempo, F., Orsini, E., Martins, L. R., Antunes, I., Lonetti, A., Chiarini, F., Tabellini, G., Evangelisti, C., Evangelisti, C., Melchionda, F., Pession, A., Bertaina, A., Locatelli, F., McCubrey, J. A., Cappellini, A., Barata, J. T., Martelli, A. M. 2014; 28 (3): 543-553


    Constitutively active casein kinase 2 (CK2) signaling is a common feature of T-cell acute lymphoblastic leukemia (T-ALL). CK2 phosphorylates PTEN (phosphatase and tensin homolog) tumor suppressor, resulting in PTEN stabilization and functional inactivation. Downregulation of PTEN activity has an impact on PI3K/Akt/mTOR signaling, which is of fundamental importance for T-ALL cell survival. These observations lend compelling weight to the application of CK2 inhibitors in the therapy of T-ALL. Here, we have analyzed the therapeutic potential of CX-4945-a novel, highly specific, orally available, ATP-competitive inhibitor of CK2α. We show that CX-4945 treatment induced apoptosis in T-ALL cell lines and patient T lymphoblasts. CX-4945 downregulated PI3K/Akt/mTOR signaling in leukemic cells. Notably, CX-4945 affected the unfolded protein response (UPR), as demonstrated by a significant decrease in the levels of the main UPR regulator GRP78/BIP, and led to apoptosis via upregulation of the ER stress/UPR cell death mediators IRE1α and CHOP. In vivo administration of CX-4945 to a subcutaneous xenotransplant model of human T-ALL significantly delayed tumor growth. Our findings indicate that modulation of the ER stress/UPR signaling through CK2 inhibition could be exploited for inducing apoptosis in T-ALL cells and that CX-4945 may be an efficient treatment for those T-ALLs displaying upregulation of CK2α/PI3K/Akt/mTOR signaling.

    View details for DOI 10.1038/leu.2013.349

    View details for Web of Science ID 000332845700007

    View details for PubMedID 24253024

  • Human Cytomegalovirus Infection Promotes Rapid Maturation of NK Cells Expressing Activating Killer Ig- like Receptor in Patients Transplanted with NKG2C(-/-) Umbilical Cord Blood JOURNAL OF IMMUNOLOGY Della Chiesa, M., Falco, M., Bertaina, A., Muccio, L., Alicata, C., Frassoni, F., Locatelli, F., Moretta, L., Moretta, A. 2014; 192 (4): 1471-1479


    NK cells are the first lymphoid population recovering after allogeneic hematopoietic stem cell transplantation and play a crucial role in early immunity after the graft. Recently, it has been shown that human CMV (HCMV) infection/reactivation can deeply influence NK cell reconstitution after umbilical cord blood transplantation by accelerating the differentiation of mature NKG2A(-) killer Ig-like receptor (KIR)(+) NK cells characterized by the expression of the NKG2C-activating receptor. In view of the hypothesis that NKG2C could be directly involved in NK cell maturation driven by HCMV infection, we analyzed the maturation and function of NK cells developing in three patients with hematological malignancies given umbilical cord blood transplantation from donors carrying a homozygous deletion of the NKG2C gene. We show that HCMV infection can drive rapid NK maturation, characterized by the expansion of CD56(dim)NKG2A(-)KIR(+) cells, even in the absence of NKG2C expression. Interestingly, this expanded mature NK cell subset expressed surface-activating KIR that could trigger NK cell cytotoxicity, degranulation, and IFN-γ release. Given the absence of NKG2C, it is conceivable that activating KIRs may play a role in the HCMV-driven NK cell maturation and that NK cells expressing activating KIRs might contribute, at least in part, to the control of infections after transplantation.

    View details for DOI 10.4049/jimmunol.1302053

    View details for Web of Science ID 000331267200016

    View details for PubMedID 24442432

  • Adoptive Immunotherapy With Antigen-Specific T Cells During Extracorporeal Membrane Oxygenation (ECMO) for Adenovirus-Related Respiratory Failure in a Child Given Haploidentical Stem Cell Transplantation PEDIATRIC BLOOD & CANCER Di Nardo, M., Li Pira, G., Amodeo, A., Cecchetti, C., Giorda, E., Ceccarelli, S., Brescia, L. P., Pirozzi, N., Rutella, S., Locatelli, F., Bertaina, A. 2014; 61 (2): 376-379


    We report on the successful infusion of human adenovirus (HAdV)-specific T cells in a child with congenital amegakaryocytic thrombocytopenia, given T-cell-depleted hematopoietic stem cell transplantation (HSCT) from the HLA-haploidentical mother during extracorporeal membrane oxygenation (ECMO) for severe HAdV-related respiratory failure. Donor-derived, interferon (IFN)-γ-secreting HAdV-specific T cells were enriched using the cytokine capture assay, after in vitro stimulation with overlapping peptides from the immunodominant HAdV5 hexon protein. Two weeks after T-cell transfer, viral load decreased and ECMO was discontinued. T-cell responses to HAdV antigens were documented after four weeks and were associated with viral clearance, immune reconstitution and clinical amelioration.

    View details for DOI 10.1002/pbc.24753

    View details for Web of Science ID 000328694300043

    View details for PubMedID 24039155

  • Impact of HCMV Infection on NK Cell Development and Function after HSCT. Frontiers in immunology Della Chiesa, M., Falco, M., Muccio, L., Bertaina, A., Locatelli, F., Moretta, A. 2013; 4: 458-?


    Natural Killer (NK) cell function is regulated by an array of inhibitory and activating surface receptors that during NK cell differentiation, at variance with T and B cells, do not require genetic rearrangement. Importantly, NK cells are the first lymphocyte population recovering after hematopoietic stem cell transplantation (HSCT). Thus, their role in early immunity after HSCT is considered crucial, as they can importantly contribute to protect the host from tumor recurrence and viral infections before T-cell immunity is fully recovered. In order to acquire effector functions and regulatory receptors, NK cell precursors undergo a maturation process that can be analyzed during immune reconstitution after HSCT. In this context, the occurrence of human cytomegalovirus (HCMV) infection/reactivation was shown to accelerate NK cell maturation by promoting the differentiation of high frequencies of NK cells characterized by a KIR(+)NKG2A(-) and NKG2C(+) mature phenotype. Thus, it appears that the development of NK cells and the distribution of NK cell receptors can be deeply influenced by HCMV infection. Moreover, in HCMV-infected subjects the emergence of so called "memory-like" or "long-lived" NK cells has been documented. These cells could play an important role in protecting from infections and maybe from relapse in patients transplanted for leukemia. All the aspects regarding the influence of HCMV infection on NK cell development will be discussed.

    View details for DOI 10.3389/fimmu.2013.00458

    View details for PubMedID 24379818

    View details for PubMedCentralID PMC3861788

  • Multiple infusions of mesenchymal stromal cells induce sustained remission in children with steroid-refractory, grade III-IV acute graft-versus-host disease BRITISH JOURNAL OF HAEMATOLOGY Ball, L. M., Bernardo, M. E., Roelofs, H., van Tol, M. J., Contoli, B., Zwaginga, J. J., Avanzini, M. A., Conforti, A., Bertaina, A., Giorgiani, G., Jol-van der Zijde, C. M., Zecca, M., Le Blanc, K., Frassoni, F., Egeler, R. M., Fibbe, W. E., Lankester, A. C., Locatelli, F. 2013; 163 (4): 501-509


    Mesenchymal stromal cell (MSC) infusions have been reported to be effective in patients with steroid-refractory, acute graft-versus-host disease (aGvHD) but comprehensive data on paediatric patients are limited. We retrospectively analysed a cohort of 37 children (aged 3 months-17 years) treated with MSCs for steroid-refractory grade III-IV aGvHD. All patients but three received multiple MSC infusions. Complete response (CR) was observed in 24 children (65%), while 13 children had either partial (n = 8) or no response (n = 5). Cumulative incidence of transplantation-related mortality (TRM) in patients who did or did not achieve CR was 17% and 69%, respectively (P = 0.001). After a median follow-up of 2.9 years, overall survival (OS) was 37%; it was 65% vs. 0% in patients who did or did not achieve CR, respectively (P = 0.001). The median time from starting steroids for GvHD treatment to first MSC infusion was 13 d (range 5-85). Children treated between 5 and 12 d after steroid initiation showed a trend for better OS (56%) and lower TRM (17%) as compared with patients receiving MSCs 13-85 d after steroids (25% and 53%, respectively; P = 0.22 and 0.06, respectively). Multiple MSC infusions are safe and effective for children with steroid-refractory aGvHD, especially when employed early in the disease course.

    View details for DOI 10.1111/bjh.12545

    View details for Web of Science ID 000326034200011

    View details for PubMedID 23992039

  • Validation of flow cytometric phospho-STAT5 as a diagnostic tool for juvenile myelomonocytic leukemia BLOOD CANCER JOURNAL Hasegawa, D., Bugarin, C., Giordan, M., BRESOLIN, S., Longoni, D., Micalizzi, C., Ramenghi, U., Bertaina, A., Basso, G., Locatelli, F., Biondi, A., te Kronnie, G., Gaipa, G. 2013; 3


    To diagnose juvenile myelomonocytic leukemia (JMML) is sometimes challenging, because around 10% of patients lack molecular abnormalities affecting Ras-MAPK (mitogen-activated protein kinase) pathway and other diseases such as cytomegalovirus infection can mimic clinical signs of JMML. In order to validate a phospho-specific flow cytometry assay assessing phospho-signal transducer and activator of transcription factor 5 (p-STAT5) as a new diagnostic tool for JMML, we examined 22 samples from children with JMML and 47 controls. CD33+/CD34+ cells from 22 patients with JMML showed hyperphosphorylation of STAT5 induced by sub-saturating doses of granulocyte-macrophage colony-stimulating factor (GM-CSF). Using a training set of samples (11 JMML and 23 controls), we identified a threshold for p-STAT5-positive after stimulation with 0.1 ng/ml GM-CSF (17.17%) that discriminates JMML from controls. This threshold was validated in an independent series (11 JMML, 24 controls and 7 cases with diseases other than JMML) where we demonstrated that patients with JMML could be distinguished from other subjects with a sensitivity of 91% (confidence interval (CI) 59-100%) and a specificity of 87% (CI 70-96%). Positive and negative predictive values were 71% (CI 42-92%) and 96% (CI 82-100%), respectively. In conclusion, flow cytometric p-STAT5 profiling is a reliable diagnostic tool for identifying patients with JMML and can contribute to consistency of current diagnostic criteria.

    View details for DOI 10.1038/bcj.2013.56

    View details for Web of Science ID 000328069400006

    View details for PubMedID 24241400

    View details for PubMedCentralID PMC3880439

  • Negative depletion of alpha/beta(+) T cells and of CD19+B lymphocytes: A novel frontier to optimize the effect of innate immunity in HLA-mismatched hematopoietic stem cell transplantation IMMUNOLOGY LETTERS Locatelli, F., Bauquet, A., Palumbo, G., Moretta, F., Bertaina, A. 2013; 155 (1-2): 21-23


    In recent years, infusion of T-cell depleted hematopoietic stem cells from an HLA-haploidentical relative has been shown to represent a suitable and effective, alternative option in patients in need of an allograft who lack an HLA-identical relative. In particular, this type of allograft is associated with the enormous advantage of offering an immediate transplant treatment to virtually all pediatric patients without an HLA-matched donor, whether related or unrelated, or a suitable umbilical cord blood unit. Several studies have shown that in patients given a T-cell depleted transplant relevant part of the anti-leukemia effect is mediated by alloreactive (i.e. KIR/HLA mismatched) Natural Killer cells originated from donor hematopoietic stem cells. After infusion of positively selected hematopoietic stem cell, fully functioning Natural Killer cells emerge in the recipient peripheral blood, persisting over time, only several weeks after the allograft. We have developed a new method of T-cell depletion (based on the physical elimination of mature T cells carrying α and β chains of the T-cell receptor), which permits to maintain mature donor-derived alloreactive Natural Killer cells and γδ(+) T cells in the graft. We, thus, started a formal study in children with hematological disorders aimed at evaluating the safety and efficacy of this approach. Preliminary results on 60 children transplanted so far after this type of graft manipulation are particularly promising.

    View details for DOI 10.1016/j.imlet.2013.09.027

    View details for Web of Science ID 000328178600007

    View details for PubMedID 24091162

  • Late respiratory failure after hematopoietic stem cells transplant: just "lung failure"? MINERVA ANESTESIOLOGICA Caldarelli, V., Gregoretti, C., Paglietti, M. G., Pavone, M., Bertaina, A., Testa, M. B., Cutrera, R. 2013; 79 (8): 969-970

    View details for Web of Science ID 000326410400020

    View details for PubMedID 23449246

  • Cellular and molecular basis of haploidentical hematopoietic stem cell transplantation in the successful treatment of high-risk leukemias: role of alloreactive NK cells. Frontiers in immunology Locatelli, F., Pende, D., Mingari, M. C., Bertaina, A., Falco, M., Moretta, A., Moretta, L. 2013; 4: 15-?


    Natural killer (NK) cells are involved in innate immune responses and play a major role in tumor surveillance and in defense against viruses. Human NK cells recognize human leukocyte antigen (HLA) class I molecules via surface receptors [killer immunoglobulin-like receptor (KIR) and NKG2A] delivering signals that inhibit NK cell function and kill HLA class I-deficient target cells, a frequent event in tumors or virus-infected cells. NK cell triggering is mediated by activating receptors that recognize ligands expressed primarily on tumors or virus-infected cells. NK cells play also a key role in the cure of high-risk leukemias. Thus, donor-derived "alloreactive" NK cells are fundamental effectors in adult acute myeloid leukemia and in pediatric acute lymphoblastic leukemia patients undergoing haploidentical hematopoietic stem cell transplantation (HSCT). Alloreactive NK cells mediate killing of leukemia cells and patient's dendritic cell, thus preventing respectively leukemic relapses and graft-vs-host responses. Cytofluorimetric analysis of KIRs expressed by NK cells allows to define the size of the alloreactive NK subset and the selection of the best potential donor. Recently, it has been shown that also the expression of activating KIRs, in particular the (C2-specific) KIR2DS1, may contribute to donor NK alloreactivity. It has also been established a correlation between the size of the alloreactive NK cell population and the clinical outcome. Notably, the alloreactive NK cells derived from donor's hematopoietic stem cells are generated and persist in patients over time. The high survival rates of patients undergoing haploidentical HSCT highlight an important new reality in the setting of allograft performed to cure otherwise fatal leukemias. Novel approaches are in progress to further improve the clinical outcome based on the infusion of donor alloreactive NK cells either as a component of the transplanted cell population or as in vitro expanded NK cells.

    View details for DOI 10.3389/fimmu.2013.00015

    View details for PubMedID 23378843

    View details for PubMedCentralID PMC3561663

  • Epigenetic Deregulation of MicroRNAs in Rhabdomyosarcoma and Neuroblastoma and Translational Perspectives INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES Romania, P., Bertaina, A., Bracaglia, G., Locatelli, F., Fruci, D., Rota, R. 2012; 13 (12): 16554-16579


    Gene expression control mediated by microRNAs and epigenetic remodeling of chromatin are interconnected processes often involved in feedback regulatory loops, which strictly guide proper tissue differentiation during embryonal development. Altered expression of microRNAs is one of the mechanisms leading to pathologic conditions, such as cancer. Several lines of evidence pointed to epigenetic alterations as responsible for aberrant microRNA expression in human cancers. Rhabdomyosarcoma and neuroblastoma are pediatric cancers derived from cells presenting features of skeletal muscle and neuronal precursors, respectively, blocked at different stages of differentiation. Consistently, tumor cells express tissue markers of origin but are unable to terminally differentiate. Several microRNAs playing a key role during tissue differentiation are often epigenetically downregulated in rhabdomyosarcoma and neuroblastoma and behave as tumor suppressors when re-expressed. Recently, inhibition of epigenetic modulators in adult tumors has provided encouraging results causing re-expression of anti-tumor master gene pathways. Thus, a similar approach could be used to correct the aberrant epigenetic regulation of microRNAs in rhabdomyosarcoma and neuroblastoma. The present review highlights the current insights on epigenetically deregulated microRNAs in rhabdomyosarcoma and neuroblastoma and their role in tumorigenesis and developmental pathways. The translational clinical implications and challenges regarding modulation of epigenetic chromatin remodeling/microRNAs interconnections are also discussed.

    View details for DOI 10.3390/ijms131216554

    View details for Web of Science ID 000312608100063

    View details for PubMedID 23443118

    View details for PubMedCentralID PMC3546707

  • Lymphocyte proliferation specific for recall, CMV and HIV antigens in miniaturized and automated format JOURNAL OF IMMUNOLOGICAL METHODS Pira, G. L., Starc, N., Conforti, A., Bertaina, A., Rutella, S., Locatelli, F., Manca, F. 2012; 384 (1-2): 135-142


    Lymphoproliferation assay (LPA) is used to test specific T-cell responses. LPA is performed in 96-well plates with 2-5×10⁵ PBMC/well. In order to test numerous antigens, as in the case of epitope mapping or screening of antigenic panels from relevant pathogens, PBMC numbers may not be sufficient. We developed a miniaturized and automated procedure to perform LPA in 384- and 1536-well plates with one fourth to one twentieth of PBMC numbers used for standard assays. Here, we demonstrate that the procedure is reliable and robust using recall antigens and protein and peptide antigens from CMV and HIV. By using HIV specific T-cell lines, we also demonstrate that sensitivity ranges overlap with those of standard LPA and that as few as 3 specific cells/well provide a positive signal. This procedure is consistent with our policy to miniaturize assays for specific T-cell immunity, as we have already established for cytokine secretion assays.

    View details for DOI 10.1016/j.jim.2012.07.022

    View details for Web of Science ID 000309572300016

    View details for PubMedID 22967924

  • Allogeneic hematopoietic stem cell transplantation in thalassemia major: results of a reduced-toxicity conditioning regimen based on the use of treosulfan BLOOD Bernardo, M. E., Piras, E., Vacca, A., Giorgiani, G., Zecca, M., Bertaina, A., Pagliara, D., Contoli, B., Pinto, R. M., Caocci, G., Mastronuzzi, A., La Nasa, G., Locatelli, F. 2012; 120 (2): 473-476


    Sixty thalassemia patients (median age, 7 years; range, 1-37) underwent allogeneic hematopoietic stem cell transplantation (HSCT) after a preparation combining thiotepa, treosulfan, and fludarabine. Before HSCT, 27 children were assigned to risk class 1 of the Pesaro classification, 17 to class 2, and 4 to class 3; 12 patients were adults. Twenty patients were transplanted from an HLA-identical sibling and 40 from an unrelated donor. The cumulative incidence of graft failure and transplantation-related mortality was 9% and 7%, respectively. Eight patients experienced grade II-IV acute GVHD, the cumulative incidence being 14%. Among 56 patients at risk, 1 developed limited chronic GVHD. With a median follow-up of 36 months (range, 4-72), the 5-year probability of survival and thalassemia-free survival are 93% and 84%, respectively. Neither the class of risk nor the donor used influenced outcome. This treosulfan-based preparation proved to be safe and effective for thalassemia patients given allogeneic HSCT.

    View details for DOI 10.1182/blood-2012-04-423822

    View details for Web of Science ID 000307412400033

    View details for PubMedID 22645178

  • Recipient CTLA-4*CT60-AA genotype is a prognostic factor for acute graft-versus-host disease in hematopoietic stem cell transplantation for thalassemia HUMAN IMMUNOLOGY Orru, S., Orru, N., Manolakos, E., Littera, R., Caocci, G., Giorgiani, G., Bertaina, A., Pagliara, D., Giardini, C., Nesci, S., Locatelli, F., Carcassi, C., La Nasa, G. 2012; 73 (3): 282-286


    Polymorphisms of the cytotoxic T-lymphocyte antigen-4 gene (CTLA-4) have been associated with autoimmune diseases and it has recently been reported that donor genotypes correlate with the outcome of allogeneic hematopoietic stem cell transplantation in leukemia patients. With the aim of confirming this finding in thalassemia patients, we investigated the influence of genotype distribution of 3 CTLA-4 gene polymorphisms in 72 thalassemia patients and their unrelated donors. A significant association was observed for recipient CT60-AA genotype and onset of grade II-IV (63.2% vs 24.5%; p = 0.001) and grade III-IV (36.4% vs 7.6%; p = 0.005) acute graft-versus-host disease (aGVHD). The same association was observed for the 88-base-pair allele of the CTLA-4 (AT)n polymorphism, which was determined to be in complete linkage disequilibrium with the CT60 A allele. Multinomial Cox regression demonstrated that this association was independent of CT60 donor genotypes or other risk factors (p = 0.016; hazard ratio = 2.8). Our data confirm that the genetic variability in CTLA-4 is an important prognostic factor for aGVHD and suggest that some of the risk factors for this complication are generated by recipient cells that persist after the myeloablative conditioning regimen.

    View details for DOI 10.1016/j.humimm.2011.12.014

    View details for Web of Science ID 000300964400011

    View details for PubMedID 22245568

    View details for PubMedCentralID PMC3314940

  • Absence of IL-12R beta 2 in CD33(+)CD38(+) pediatric acute myeloid leukemia cells favours progression in NOD/SCID/IL2R gamma C-deficient mice LEUKEMIA Ferretti, E., Montagna, D., Di Carlo, E., Cocco, C., Ribatti, D., Ognio, E., Sorrentino, C., Lisini, D., Bertaina, A., Locatelli, F., Pistoia, V., Airoldi, I. 2012; 26 (2): 225-235


    Childhood acute myeloid leukemia (AML) is a hematological malignancy in which tumor burden is continuously replenished by leukemic-initiating cells (ICs), which proliferate slowly and are refractory to chemotherapeutic agents. We investigated whether interleukin (IL)-12, an immuno-modulatory cytokine with anti-tumor activity, may target AML blasts (CD45(+)CD33(+)) and populations known to contain leukemia ICs (that is, CD34(+)CD38(-), CD33(+)CD38(+) and CD44(+)CD38(-) cells). We demonstrate for the first time that: i) AML blasts and their CD34(+)CD38(-), CD33(+)CD38(+), CD44(+)CD38(-) subsets express the heterodimeric IL-12 receptor (IL-12R), ii) AML cells injected subcutaneously into NOD/SCID/Il2rg(-/-) (NSG) mice developed a localized tumor mass containing leukemic ICs and blasts that were virtually eliminated by IL-12 treatment, iii) AML cells injected intravenously into NSG mice engrafted within the first month in the spleen, but not in bone marrow or peripheral blood. At this time, IL-12 dramatically dampened AML CD45(+)CD33(+), CD34(+)CD38(-), CD33(+)CD38(+) and CD44(+)CD38(-) populations, only sparing residual CD33(+)CD38(+) cells that did not express IL-12Rβ2. From 30 to 60 days after the initial inoculum, these IL-12-unresponsive cells expanded and metastasized in both control and IL-12-treated NSG mice. Our data indicate that the absence of IL-12Rβ2 in pediatric AML cells favours leukemia progression in NOD/SCID/IL2Rγc-deficient mice.

    View details for DOI 10.1038/leu.2011.213

    View details for Web of Science ID 000300419100004

    View details for PubMedID 21844875

  • Interactions between killer immunoglobulin-like receptors and their human leucocyte antigen Class I ligands influence the outcome of unrelated haematopoietic stem cell transplantation for thalassaemia: a novel predictive algorithm BRITISH JOURNAL OF HAEMATOLOGY Littera, R., Orru, N., Caocci, G., Sanna, M., Mulargia, M., Piras, E., Vacca, A., Giardini, C., Orofino, M. G., Visani, G., Bertaina, A., Giorgiani, G., Locatelli, F., Carcassi, C., La Nasa, G. 2012; 156 (1): 118-128


    In a study conducted on 114 patients undergoing unrelated donor haematopoietic stem cell transplantation (HSCT) for thalassaemia, we observed that the lack of activating killer immunoglobulin-like receptors (KIRs) on donor natural killer (NK) cells significantly increased the risk of graft-versus-host disease (GvHD) [hazard risk (HR) 4.2, 95% confidence interval (CI) 1.7-10.1, P = 0.002] and transplantation-related mortality (HR 4.7, 95% CI 1.6-14.2, P = 0.01). The risk of GvHD furthermore increased when recipients heterozygous for HLA-C KIR ligand groups (C1/C2) were transplanted from donors completely lacking activating KIRs (HR 6.1, 95% CI 1.9-19.2, P = 0.002). We also found that the risk of rejection was highest when the recipient was homozygous for the C2 HLA-KIR ligand group and the donor carried two or more activating KIRs (HR 6.8, 95% CI 1.9-24.4, P = 0.005). By interpolating the number of donor activating KIRs with recipient HLA-C KIR ligands, we created an algorithm capable of stratifying patients according to the immunogenetic risk of complications following unrelated HSCT. In clinical practice, this predictive tool could serve as an important supplement to clinical judgement and decision-making.

    View details for DOI 10.1111/j.1365-2141.2011.08923.x

    View details for Web of Science ID 000298058100013

    View details for PubMedID 22077388

  • Strategies to optimize the outcome of children given T-cell depleted HLA-haploidentical hematopoietic stem cell transplantation BEST PRACTICE & RESEARCH CLINICAL HAEMATOLOGY Locatelli, F., Vinti, L., Palumbo, G., Rossi, F., Bertaina, A., Mastronuzzi, A., Bernardo, M. E., Rutella, S., Dellabona, P., Giorgiani, G., Moretta, A., Moretta, L. 2011; 24 (3): 339-349


    The most advanced frontier of allogeneic hematopoietic stem cell transplantation (allo-HSCT) is represented by the use of an HLA-partially matched relative as donor. In this type of transplantation, donor-derived natural killer (NK) cells, which are alloreactive toward recipient cells, significantly contribute to the eradication of leukemia blasts. Alloreactive NK cells may also kill host dendritic cells and T lymphocytes, thus preventing graft-versus-host disease and graft rejection, respectively. Sophisticated strategies of adoptive infusion of T-cell lines/clones specific for the most life-threatening pathogens (namely cytomegalovirus, Epstein-Barr virus, Aspergillus and Adenovirus) have been envisaged, and successfully tested in a few pilot trials, to protect the recipient in the early post-transplantation period. In these patients, also ex-vivo expanded mesenchymal stromal cells have been shown to be beneficial for preventing graft failure. Novel and effective strategies aimed at further augmenting the graft-versus-leukemia effect and at optimizing prevention/treatment of opportunistic/viral infections are warranted.

    View details for DOI 10.1016/j.beha.2011.04.004

    View details for Web of Science ID 000295655400004

    View details for PubMedID 21925087

  • Co-infusion of ex vivo-expanded, parental MSCs prevents life-threatening acute GVHD, but does not reduce the risk of graft failure in pediatric patients undergoing allogeneic umbilical cord blood transplantation BONE MARROW TRANSPLANTATION Bernardo, M. E., Ball, L. M., Cometa, A. M., Roelofs, H., Zecca, M., Avanzini, M. A., Bertaina, A., Vinti, L., Lankester, A., Maccario, R., Ringden, O., Le Blanc, K., Egeler, R. M., Fibbe, W. E., Locatelli, F. 2011; 46 (2): 200-207


    When compared with BMT, umbilical cord blood transplantation (UCBT) is associated with a lower rate of engraftment and delayed hematological/immunological recovery. This leads to increased risk of TRM in the early post transplantation period due to infection. Acute GVHD, although occurring less frequently in UCBT compared with BMT, is also significantly associated with increased rate of early TRM. BM MSCs are known to support normal in vivo hematopoiesis, and co-transplantation of MSCs has been shown to enhance engraftment of human cord blood hematopoietic cells in nonobese diabetic/SCID mice. In 13 children with hematological disorders (median age 2 years) undergoing UCBT, we co-transplanted paternal, HLA-disparate MSCs with the aim of improving hematological recovery and reducing rejection. We observed no differences in hematological recovery or rejection rates compared with 39 matched historical controls, most of whom received G-CSF after UCBT. However, the rate of grade III and IV acute GVHD was significantly decreased in the study cohort when compared with controls (P=0.05), thus resulting in reduced early TRM. Although these data do not support the use of MSCs in UCBT to support hematopoietic engraftment, they suggest that MSCs, possibly because of their immunosuppressive effect, may abrogate life-threatening acute GVHD and reduce early TRM.

    View details for DOI 10.1038/bmt.2010.87

    View details for Web of Science ID 000287190700005

    View details for PubMedID 20400983

  • The role of killer immunoglobulin-like receptor haplotypes on the outcome of unrelated donor haematopoietic SCT for thalassaemia BONE MARROW TRANSPLANTATION Littera, R., Orru, N., Vacca, A., Bertaina, A., Caocci, G., Mulargia, M., Giardini, C., Piras, E., Mastronuzzi, A., Vinti, L., Orru, S., Locatelli, F., Carcassi, C., La Nasa, G. 2010; 45 (11): 1618-1624


    Recent insight into the pathophysiology of acute GVHD after allogeneic haematopoietic SCT has led to a growing interest in the role of natural killer (NK) cells. NK cell cytotoxicity is mainly regulated by the interaction of activating and inhibitory killer immunoglobulin-like receptors (KIRs) with their respective ligands. To investigate the impact of KIRs and their ligands on haematopoietic SCT outcome, we performed a retrospective study of 78 transfusion-dependent thalassaemia patients (median age 10 years, range 1-29 years) transplanted from an unrelated donor selected using high-resolution molecular typing for both class I and II loci after a myeloablative conditioning regimen. GVHD prophylaxis consisted of CsA, short-term MTX and anti-thymocyte globulin in all patients. We found that patients transplanted from donors homozygous for KIR haplotype A had a greater risk of developing grade II-IV acute GVHD compared with those transplanted from a donor carrying at least one B haplotype (hazard ratio=4.5, 99% confidence interval=1.2-17.1, P=0.003). Our study suggests that KIR genotyping of donor and recipient pairs could contribute to the identification of patients at high risk for developing severe complications of haematopoietic SCT and thus may help with the choice of intensity of GVHD prophylaxis.

    View details for DOI 10.1038/bmt.2010.24

    View details for Web of Science ID 000284063500007

    View details for PubMedID 20173792

  • Reconstitution of repertoire of natural killer cell receptors after transplantation: just a question of time? BONE MARROW TRANSPLANTATION Locatelli, F., Bertaina, A. 2010; 45 (6): 968-969

    View details for DOI 10.1038/bmt.2010.116

    View details for Web of Science ID 000278573600002

    View details for PubMedID 20531444

  • Cord blood transplantation in children with haematological malignancies BEST PRACTICE & RESEARCH CLINICAL HAEMATOLOGY Bertaina, A., Bernardo, M. E., Caniglia, M., Vinti, L., Giorgiani, G., Locatelli, F. 2010; 23 (2): 189-196


    Umbilical cord blood transplantation (UCBT) is largely used to treat children affected by haematological malignant disorders. In comparison to bone marrow transplantation (BMT), advantages of UCBT include lower incidence and severity of graft-versus-host disease, easier procurement and prompter availability of cord blood cells, and the possibility of using donors having HLA disparities with the recipient. The large experience accumulated so far has shown that UCBT offers to children a probability of cure at least comparable to that of patients transplanted with bone marrow cells. Since it has been demonstrated that an inverse correlation between the number of nucleated cord blood cells infused per kg recipient body weight and the risk of dying for transplantation-related causes exists, recently developed strategies aimed at increasing the number of cord blood progenitors and at favouring stem cell homing could further optimize the outcome of children with leukemia or other malignancies receiving UCBT.

    View details for DOI 10.1016/j.beha.2010.06.005

    View details for Web of Science ID 000282559800004

    View details for PubMedID 20837330

  • Cord blood transplantation in patients with hemoglobinopathies TRANSFUSION AND APHERESIS SCIENCE Boncimino, A., Bertaina, A., Locatelli, F. 2010; 42 (3): 277-281


    Despite the optimization of conventional treatment, both thalassemia and sickle cell disease are still associated with significant morbidity and mortality, especially in developing countries. Allogeneic transplantation of hematopoietic progenitors is the only curative treatment and represents an attractive option for these patients. In view of the low incidence of graft-versus-host disease associated with the procedure, allogeneic cord blood transplantation (CBT) is particularly appealing for patients with non-malignant disorders. Available evidence indicates that related donor CBT is a safe and effective option for patients with hemoglobinopathies, able to offer results at least as good as those reported using bone marrow cells.

    View details for DOI 10.1016/j.transci.2010.03.006

    View details for Web of Science ID 000279531900010

    View details for PubMedID 20382570

  • Comparison between an artificial neural network and logistic regression in predicting acute graft-vs-host disease after unrelated donor hematopoietic stem cell transplantation in thalassemia patients EXPERIMENTAL HEMATOLOGY Caocci, G., Baccoli, R., Vacca, A., Mastronuzzi, A., Bertaina, A., Piras, E., Littera, R., Locatelli, F., Carcassi, C., La Nasa, G. 2010; 38 (5): 426-433


    There is growing interest in the development of prognostic models for predicting the occurrence of acute graft-vs-host disease (aGVHD) after unrelated donor hematopoietic stem cell transplantation. A high number of variables have been shown to play a role in aGVHD, but the search for a predictive algorithm is still ongoing. Artificial neural networks (ANNs) represent an attractive alternative to multivariate analysis for clinical prognosis. So far, no reports have investigated the ability of ANNs in predicting HSCT outcome.We compared the prognostic performance of ANNs with that of logistic regression (LR) in 78 beta-thalassemia major patients given unrelated donor hematopoietic stem cell transplantation. Twenty-four independent variables were analyzed for their potential impact on outcomes.Twenty-six patients (33.3%) developed grade II to IV aGVHD. In multivariate analysis, homozygosity for donor KIR haplotype A (p = 0.03), donor age (p = 0.05), and donor homozygosity for the deletion of the human leukocyte antigen-G 14-bp polymorphism (p = 0.05) were independently significantly correlated to aGVHD. The mean sensitivity of LR and ANNs (capability of predicting aGVHD in patients who developed aGVHD) in test datasets was 21.7% and 83.3%, respectively (p < 0.001); the mean specificity (capability of predicting absence of aGVHD in patients who did not develop aGVHD) was 80.5% and 90.1%, respectively (p = NS).Although ANNs are unable to calculate the weight of single variables on outcomes, they were found to have a better performance than LR. A combination of these two methods could be more efficient in predicting outcomes and help tailor GVHD prophylaxis regimens according to the predicted risk of each patient. Whether ANN technology will provide better predictive performance when applied to other datasets remains to be confirmed.

    View details for DOI 10.1016/j.exphem.2010.02.012

    View details for Web of Science ID 000277015400009

    View details for PubMedID 20206661

  • The role of reduced intensity preparative regimens in patients with thalassemia given hematopoietic transplantation COOLEY'S ANEMIA: NINTH SYMPOSIUM Bertaina, A., Bernardo, M. E., Mastronuzzi, A., La Nasa, G., Locatelli, F. 2010; 1202: 141-148


    Allogeneic hematopoietic stem cell transplantation (HSCT) still remains the only curative treatment for patients with thalassemia major (TM). However, HSCT is associated with a non-negligible risk of both transplantation-related mortality (TRM) and morbidity. Great interest and relevant expectations have been raised by the introduction in the clinical practice of reduced-intensity preparative regimens, which may represent an effective strategy to reduce the toxicity of transplantation and may also help reduce the incidence of late effects. Although some reports have documented the feasibility of using reduced-intensity preparative regimens for successfully treating patients with TM, a high incidence of graft failure has been frequently reported. Recently, treosulfan-based myeloablation has been demonstrated to be associated with limited extra-medullary toxicity and a high rate of sustained donor engraftment. This novel approach is a promising alternative for reducing the risk of life-threatening complications and increasing the number of TM patients successfully cured with an allograft.

    View details for DOI 10.1111/j.1749-6632.2010.05590.x

    View details for Web of Science ID 000283099600022

    View details for PubMedID 20712785

  • B LYMPHOCYTE SUBSETS AND THEIR FUNCTIONAL ACTIVITY IN THE EARLY MONTHS OF LIFE INTERNATIONAL JOURNAL OF IMMUNOPATHOLOGY AND PHARMACOLOGY Avanzini, M. A., Maccario, R., Belloni, C., Carrera, G., Bertaina, A., Cagliuso, M., La Rocca, M., Valsecchi, C., Mantelli, M., Castellazzi, A. M., Quinti, I., De Silvestri, A., Marconi, M. 2010; 23 (1): 247-254


    In the present study we evaluated B-cell subsets and their functional development in 74 newborns from birth to 6 months of life. Moreover, we evaluated natural antibody production in vitro. The results documented a predominance of naive B-lymphocytes at all time-points evaluated, decreasing from birth to 6 months (p=0.009). The percentages of CD27+IgD+ and CD27+IgDneg memory B-cells were very low at birth and significantly increased only at 6 months (p=0.02 and p less than 0.001, respectively). We found a significant increase only in in vitro stimulated IgG production at 6 months as compared to birth (p less than 0.001). Moreover, a lower secretion of anti-Pn IgM antibodies up to 6 months of age, as compared to controls was observed. Our results underline that the susceptibility and severe course of infection in the neonate can be attributed, at least in part, to the lack of pre-existing immunological memory and competent adaptive immunity.

    View details for Web of Science ID 000276758600022

    View details for PubMedID 20378010

  • Clofarabine, cyclophosphamide and etoposide as single-course re-induction therapy for children with refractory/multiple relapsed acute lymphoblastic leukaemia BRITISH JOURNAL OF HAEMATOLOGY Locatelli, F., Testi, A. M., Bernardo, M. E., Rizzari, C., Bertaina, A., Merli, P., Pession, A., Giraldi, E., Parasole, R., Barberi, W., Zecca, M. 2009; 147 (3): 371-378


    The safety and efficacy of the combination clofarabine/cyclophosphamide/etoposide were evaluated in children with advanced acute lymphoblastic leukaemia (ALL). The study enrolled 25 paediatric patients (median age 12.5 years) with either refractory (n = 17; 68%) or multiple relapsed (n = 8; 32%) ALL to receive clofarabine 40 mg/m(2), cyclophosphamide 400 mg/m(2) and etoposide 150 mg/m(2), daily for 5 consecutive days. No patient died from treatment-related complications. The most common adverse events were febrile neutropenia, mucositis and reversible liver toxicity; no case of liver veno-occlusive disease was reported. The overall remission rate was 56%: 13 patients (52%) achieved complete remission (CR) and one (4%) CR without platelet recovery (CRp). In seven of the 13 (54%) patients achieving CR, remissions were of sufficient duration to allow patients to receive allogeneic haematopoietic stem cell transplantation. The probability of CR/CRp was greater in the 17 patients with B cell precursor ALL than in the eight with T-ALL (76% vs. 12%, respectively, P < 0.01). The 18-month overall survival probability was 39% and 0% in patients who did or did not respond to the treatment, respectively (P < 0.01). These data suggest that the clofarabine/cyclophosphamide/etoposide regimen is well tolerated and can induce clinical response in a relevant proportion of children with refractory/multiple relapsed ALL.

    View details for DOI 10.1111/j.1365-2141.2009.07882.x

    View details for Web of Science ID 000270727900011

    View details for PubMedID 19747360

  • Transplantation and innate immunity: the lesson of natural killer cells ITALIAN JOURNAL OF PEDIATRICS Bertaina, A., Locatelli, F., Moretta, L. 2009; 35


    Natural killer cells have been demonstrated to play a major role in mediating an anti-leukemia effect in patients given a T-cell depleted allogeneic hematopoietic stem cell transplantation from an HLA-haploidentical family donor. In particular, donor-derived natural killer cells, which are alloreactive (i.e. KIR/HLA mismatched) towards recipient cells, significantly contribute to the eradication of leukemia blasts escaping the preparative regimen to transplantation. A recent study on high-risk pediatric acute lymphoblastic leukemia refractory to chemotherapy further highlighted the importance of donors with alloreactive natural killer cells in haploidentical hematopoietic stem cell transplantation, as it demonstrated that these cells can emerge starting from the fourth-fifth month after the allograft and persist for many months. This study represents a major breakthrough in the cure of otherwise fatal leukemias, providing information on the best criteria for choosing the optimal donor.

    View details for DOI 10.1186/1824-7288-35-44

    View details for Web of Science ID 000208014800043

    View details for PubMedID 20076779

    View details for PubMedCentralID PMC2806872

  • Human respiratory syncytial virus (hRSV) RNA quantification in nasopharyngeal secretions identifies the hRSV etiologic role in acute respiratory tract infections of hospitalized infants JOURNAL OF CLINICAL VIROLOGY Carnpanini, G., Percivalle, E., Baldanti, F., Rovida, F., Bertaina, A., Marchi, A., Stronati, M., Gerna, G. 2007; 39 (2): 119-124


    Human respiratory syncytial virus (hRSV) detection in nasopharyngeal aspirates (NPAs) from infants with acute respiratory tract infection (ARTI) does not prove the hRSV etiology of the current ARTI episode. HRSV RNA quantification may help in affording this issue.hRSV was detected by quantitative reverse transcription-PCR in NPAs taken upon admission to hospital and, whenever possible, at discharge and subsequent medical visits.Prospective study, including 63 infants affected by either hRSV upper or lower ARTI.Based on the kinetics of viral load, hRSV etiology was identified in 25 infants in whom hRSV load dropped from 2.5 x 10(6) upon admission (presence of respiratory symptoms) to 7.5 x 10(2)RNAcopies/ml NPA upon discharge (absence of symptoms) after a median time of 5 days, and in 19 infants, in whom hRSV load was determined at admission only, in association with clinical symptoms (2.4 x 10(6)copies/ml). Furthermore, low levels of hRSV RNA (<1 x 10(5)copies/ml NPA) identified 14 patients with non-hRSV ARTI. Finally, in 14 infants with hRSV coinfections or sequential infections, hRSV quantification defined the hRSV role in the current ARTI episode.hRSV RNA quantification is critical in defining the hRSV role in respiratory infections.

    View details for DOI 10.1016/j.jcv.2007.03.009

    View details for Web of Science ID 000247509500009

    View details for PubMedID 17452001