Sasa Vasilijic

All Publications

• Identification of immune-related candidate biomarkers in plasma of patients with sporadic vestibular schwannoma. Science advances Vasilijic, S., Atai, N. A., Hyakusoku, H., Worthington, S., Ren, Y., Sagers, J. E., Sahin, M. I., Brown, A., Reddy, R., Malhotra, C., Fujita, T., Landegger, L. D., Lewis, R., Welling, D. B., Stankovic, K. M. 2023; 9 (45): eadf7295

  Abstract

  Vestibular schwannoma (VS) is an intracranial tumor arising from neoplastic Schwann cells and typically presenting with hearing loss. The traditional belief that hearing deficit is caused by physical expansion of the VS, compressing the auditory nerve, does not explain the common clinical finding that patients with small tumors can have profound hearing loss, suggesting that tumor-secreted factors could influence hearing ability in VS patients. We conducted profiling of patients' plasma for 66 immune-related factors in patients with sporadic VS (N > 170) and identified and validated candidate biomarkers associated with tumor size (S100B) and hearing (MCP-3). We further identified a nine-biomarker panel (TNR-R2, MIF, CD30, MCP-3, IL-2R, BLC, TWEAK, eotaxin, and S100B) with outstanding discriminatory ability for VS. These findings revealed possible therapeutic targets for VS, providing a unique diagnostic tool that may predict hearing change and tumor growth in VS patients, and may inform the timing of tumor resection to preserve hearing.

  View details for DOI 10.1126/sciadv.adf7295

  View details for PubMedID 37948527

  View details for PubMedCentralID PMC10637750

• Immune Profiling of Secreted Factors from Human Vestibular Schwannoma Cells and Tumor-associated Macrophages. The Laryngoscope Stankovic, K. M., Batts, S., Welling, D. B., Vasilijic, S. 2023

  Abstract

  This study compared the immune-related secretory capacity of human vestibular schwannoma (VS) and tumor-assisted macrophages (TAMs) with their normal counterparts (Schwann cells [SC] and peripheral blood monocyte-derived macrophages [Mo-MFs], respectively), and examined relationships with presurgical hearing and tumor size.VS tumors (n = 16), auditory nerve (n = 1), blood (n = 9), and great auricular nerves (n = 3) were used. SCs (S100B+ ) and TAMs (CD68+ ) were isolated from VS tissue for culture. The secreted levels of 65 immune-related factors were measured and compared using unpaired t-tests with Welch correction (schwannoma vs. SCs) or Mann-Whitney tests (TAMs and Mo-MFs). Associations between factor concentration and word recognition (WR), pure-tone average (PTA), and tumor size were evaluated with Spearman correlation.Secreted factors with significantly higher concentrations in schwannoma versus SC supernatants included IL-2 and BAFF, whereas MMP-1, IL-6, FGF-2, VEGF-A, MIP-3α, and GRO-α concentrations were significantly higher in TAMs versus Mo-MFs (all p < 0.05). Worse WR was significantly associated with higher secretion of fractalkine, eotaxin-3, CD30, and IL-16 by VS cells; IP-10, eotaxin-3, multiple interleukins, GM-CSF, SCF, and CD30 by TAMs; and TNF-α and MIP-1α by Mo-MFs (all p < 0.05). Worse PTA was significantly correlated with higher secretion of IL-16 by VS cells (p < 0.05). Larger tumor size was significantly correlated with higher secretion of eotaxin by VS cells, and of IL-7, IL-21, and LIF by TAMs (all p = 0.017).Differential secretion of immune-related factors was observed in schwannoma versus normal SCs and in TAMs versus Mo-MFs, some of which were correlated with worse hearing and larger VS tumors.N/A Laryngoscope, 2023.

  View details for DOI 10.1002/lary.31067

  View details for PubMedID 37776249

• Orally administered fluorescent nanosized polystyrene particles affect cell viability, hormonal and inflammatory profile, and behavior in treated mice. Environmental pollution (Barking, Essex : 1987) Nikolic, S., Gazdic-Jankovic, M., Rosic, G., Miletic-Kovacevic, M., Jovicic, N., Nestorovic, N., Stojkovic, P., Filipovic, N., Milosevic-Djordjevic, O., Selakovic, D., Zivanovic, M., Seklic, D., Milivojevic, N., Markovic, A., Seist, R., Vasilijic, S., Stankovic, K. M., Stojkovic, M., Ljujic, B. 2022: 119206

  Abstract

  Commercially manufactured or generated through environmental degradation, microplastics (MPs) and nanoplastics (NPs) considerably contribute to environmental pollution. There is a knowledge gap in how exposure to MPs/NPs changes cellular function and affects animal and human health. Here, we demonstrate that after oral uptake, fluorescent polystyrene (PS) nanoparticles pass through the mouse digestive system, accumulate and aggregate in different organs, and induce functional changes in cells and organs. Using cochlear explant as a novel in vitro system, we confirmed the consequences of PS-MP/NP interaction with inner ear cells by detecting aggregates and hetero-aggregates of PS particles in hair cells. The testes of treated males accumulated MPs/NPs in the interstitial compartment surrounding the seminiferous tubules, which was associated with a statistically significant decrease in testosterone levels. Male mice showed increased secretion of interleukins (IL-12p35 and IL-23) by splenocytes while cyto- and genotoxicity tests indicated impaired cell viability and increased DNA damage in spleen tissue. Males also showed a broad range of anxiogenic responses to PS nanoparticles while hippocampal samples from treated females showed an increased expression of Bax and Nlrp3 genes, indicating a pro-apoptotic/proinflammatory effect of PS treatment. Taken together, induced PS effects are also gender-dependent, and therefore, strongly motivate future research to mitigate the deleterious effects of nanosized plastic particles.

  View details for DOI 10.1016/j.envpol.2022.119206

  View details for PubMedID 35405220

• Sporadic Vestibular Schwannoma Size and Location Do not Correlate With the Severity of Hearing Loss at Initial Presentation. Frontiers in oncology Brown, A., Early, S., Vasilijic, S., Stankovic, K. M. 2022; 12: 836504

  Abstract

  Vestibular schwannoma (VS) is a non-malignant intracranial neoplasm arising from the vestibular branch of the 8th cranial nerve; sensorineural hearing loss (SNHL) is the most common associated symptom. Understanding whether VS imaging characteristics at the time of VS diagnosis can be associated with severity of VS-induced SNHL can impact patient counseling and define promising areas for future research. Patients diagnosed with VS at Massachusetts Eye and Ear (MEE) from 1994 through 2018 were analyzed if magnetic resonance imaging at VS presentation and sequential audiometry were available. Results were compared with original studies available in PubMed, written in English, on VS imaging characteristics and their impact on hearing in patients. A total of 477 patients with unilateral VS from the MEE database demonstrated no significant correlation between any features of tumor imaging at the time of VS diagnosis, such as VS size, impaction or location, and any hearing loss metric. Twenty-three published studies on the impact of VS imaging characteristics on patient hearing met inclusion criteria, with six solely involving NF2 patients and three including both sporadic and NF2-related VS patients. Fifteen studies reported a significant relationship between SNHL and at least one VS imaging characteristic; however, these trends were universally limited to NF2 patients or involved small patient populations, and were not reproduced in larger studies. Taken together, SNHL in sporadic VS patients is not readily associated solely with any tumor imaging characteristics. This finding motivates future studies to define how VS microenvironment and secreted molecules influence VS-induced SNHL.

  View details for DOI 10.3389/fonc.2022.836504

  View details for PubMedID 35372070

• Nanodesigned coatings obtained by plasma electrolytic oxidation of titanium implant and their cytotoxicity JOURNAL OF APPLIED BIOMATERIALS & FUNCTIONAL MATERIALS Magic, M., Colovic, B., Vasilijic, S., Tadic, N., Stojadinovic, S., Jokanovic, V. 2021; 19: 2280800018822252

  Abstract

  The titanium implant was treated with plasma electrolytic oxidation and subsequent ionic exchange and thermal treatment in order to obtain bioactive layer consisting of titanium oxide, calcium and sodium titanates and hydroxyapatite, as confirmed by X-ray diffraction (XRD). Scanning electron microscopy (SEM) revealed that the given method, besides corresponding phase composition, enables suitable nanotopology for cell attachment and proliferation. Cytotoxicity investigations by MTT, LDH and propidium iodide assays and light microscopy showed that these coatings were not toxic to L929 cells.

  View details for DOI 10.1177/2280800018822252

  View details for Web of Science ID 000720310600001

  View details for PubMedID 34783619

• Losartan prevents tumor-induced hearing loss and augments radiation efficacy in NF2 schwannoma rodent models. Science translational medicine Wu, L., Vasilijic, S., Sun, Y., Chen, J., Landegger, L. D., Zhang, Y., Zhou, W., Ren, J., Early, S., Yin, Z., Ho, W. W., Zhang, N., Gao, X., Lee, G. Y., Datta, M., Sagers, J. E., Brown, A., Muzikansky, A., Stemmer-Rachamimov, A., Zhang, L., Plotkin, S. R., Jain, R. K., Stankovic, K. M., Xu, L. 2021; 13 (602)

  Abstract

  Hearing loss is one of the most common symptoms of neurofibromatosis type 2 (NF2) caused by vestibular schwannomas (VSs). Fibrosis in the VS tumor microenvironment (TME) is associated with hearing loss in patients with NF2. We hypothesized that reducing the fibrosis using losartan, an FDA-approved antihypertensive drug that blocks fibrotic and inflammatory signaling, could improve hearing. Using NF2 mouse models, we found that losartan treatment normalized the TME by (i) reducing neuroinflammatory IL-6/STAT3 signaling and preventing hearing loss, (ii) normalizing tumor vasculature and alleviating neuro-edema, and (iii) increasing oxygen delivery and enhancing efficacy of radiation therapy. In preparation to translate these exciting findings into the clinic, we used patient samples and data and demonstrated that IL-6/STAT3 signaling inversely associated with hearing function, that elevated production of tumor-derived IL-6 was associated with reduced viability of cochlear sensory cells and neurons in ex vivo organotypic cochlear cultures, and that patients receiving angiotensin receptor blockers have no progression in VS-induced hearing loss compared with patients on other or no antihypertensives based on a retrospective analysis of patients with VS and hypertension. Our study provides the rationale and critical data for a prospective clinical trial of losartan in patients with VS.

  View details for DOI 10.1126/scitranslmed.abd4816

  View details for PubMedID 34261799

• Cochlin Deficiency Protects Against Noise-Induced Hearing Loss FRONTIERS IN MOLECULAR NEUROSCIENCE Seist, R., Landegger, L. D., Robertson, N. G., Vasilijic, S., Morton, C. C., Stankovic, K. M. 2021; 14: 670013

  Abstract

  Cochlin is the most abundant protein in the inner ear. To study its function in response to noise trauma, we exposed adolescent wild-type (Coch +/+ ) and cochlin knock-out (Coch -/-) mice to noise (8-16 kHz, 103 dB SPL, 2 h) that causes a permanent threshold shift and hair cell loss. Two weeks after noise exposure, Coch-/- mice had substantially less elevation in noise-induced auditory thresholds and hair cell loss than Coch + / + mice, consistent with cochlin deficiency providing protection from noise trauma. Comparison of pre-noise exposure thresholds of auditory brain stem responses (ABRs) and distortion product otoacoustic emissions (DPOAEs) in Coch-/- mice and Coch + / + littermates revealed a small and significant elevation in thresholds of Coch-/- mice, overall consistent with a small conductive hearing loss in Coch-/- mice. We show quantitatively that the pro-inflammatory component of cochlin, LCCL, is upregulated after noise exposure in perilymph of wild-type mice compared to unexposed mice, as is the enzyme catalyzing LCCL release, aggrecanase1, encoded by Adamts4. We further show that upregulation of pro-inflammatory cytokines in perilymph and cochlear soft-tissue after noise exposure is lower in cochlin knock-out than wild-type mice. Taken together, our data demonstrate for the first time that cochlin deficiency results in conductive hearing loss that protects against physiologic and molecular effects of noise trauma.

  View details for DOI 10.3389/fnmol.2021.670013

  View details for Web of Science ID 000658161600001

  View details for PubMedID 34108864

  View details for PubMedCentralID PMC8180578

• Postnatal expression and possible function of RANK and RANKL in the murine inner ear BONE Kao, S., Katsumi, S., Han, D., Bizaki-Vallaskangas, A. J., Vasilijic, S., Landegger, L. D., Kristiansen, A. G., McKenna, M. J., Stankovic, K. M. 2021; 145: 115837

  Abstract

  The bone encasing the inner ear, known as the otic capsule, is unique because it remodels little postnatally compared to other bones in the body. Previous studies established that osteoprotegerin (OPG) in the inner ear inhibits otic capsule remodeling. OPG acts as a decoy receptor of receptor activator of nuclear factor κB ligand (RANKL) to disrupt the interaction between RANKL and RANK, the primary regulators of bone metabolism. Here we studied the expression and function of RANK and RANKL in the murine cochlea. Using a combination of in situ hybridization, real-time quantitative RT-PCR, and western blot, we demonstrate that Rankl and Rank genes and their protein products are expressed in the intracochlear soft tissues and the otic capsule in a developmentally regulated manner. Using a culture of neonatal murine cochlear neurons, we show that the interaction between RANK and RANKL inhibits neurite outgrowth in these neurons, and is associated with upregulation of NOGO-A expression. Taken together, our results suggest that, in addition to regulating otic capsule bone remodeling, RANK and RANKL expressed by intracochlear soft tissues may also regulate spiral ganglion neuron function by affecting neurite outgrowth.

  View details for DOI 10.1016/j.bone.2020.115837

  View details for Web of Science ID 000623120700009

  View details for PubMedID 33385614

• New developments in neurofibromatosis type 2 and vestibular schwannoma. Neuro-oncology advances Ren, Y., Chari, D. A., Vasilijic, S., Welling, D. B., Stankovic, K. M. 2020; 3 (1): vdaa153

  Abstract

  Neurofibromatosis type 2 (NF2) is a rare autosomal dominant disorder characterized by the development of multiple nervous system tumors due to mutation in the NF2 tumor suppressor gene. The hallmark feature of the NF2 syndrome is the development of bilateral vestibular schwannomas (VS). Although there is nearly 100% penetrance by 60 years of age, some patients suffer from a severe form of the disease and develop multiple tumors at an early age, while others are asymptomatic until later in life. Management options for VS include surgery, stereotactic radiation, and observation with serial imaging; however, currently, there are no FDA-approved pharmacotherapies for NF2 or VS. Recent advancements in the molecular biology underlying NF2 have led to a better understanding of the etiology and pathogenesis of VS. These novel signaling pathways may be used to identify targeted therapies for these tumors. This review discusses the clinical features and treatment options for sporadic- and NF2-associated VS, the diagnostic and screening criteria, completed and ongoing clinical trials, quality of life metrics, and opportunities for future research.

  View details for DOI 10.1093/noajnl/vdaa153

  View details for PubMedID 33604573

  View details for PubMedCentralID PMC7881257

• Regeneration of Cochlear Synapses by Systemic Administration of a Bisphosphonate FRONTIERS IN MOLECULAR NEUROSCIENCE Seist, R., Tong, M., Landegger, L. D., Vasilijic, S., Hyakusoku, H., Katsumi, S., McKenna, C. E., Edge, A. B., Stankovic, K. M. 2020; 13: 87

  Abstract

  Sensorineural hearing loss (SNHL) caused by noise exposure and attendant loss of glutamatergic synapses between cochlear spiral ganglion neurons (SGNs) and hair cells is the most common sensory deficit worldwide. We show here that systemic administration of a bisphosphonate to mice 24 h after synaptopathic noise exposure regenerated synapses between inner hair cells and SGNs and restored cochlear function. We further demonstrate that this effect is mediated by inhibition of the mevalonate pathway. These results are highly significant because they suggest that bisphosphonates could reverse cochlear synaptopathy for the treatment of SNHL.

  View details for DOI 10.3389/fnmol.2020.00087

  View details for Web of Science ID 000556565200001

  View details for PubMedID 32765216

  View details for PubMedCentralID PMC7381223

• Combination therapy with mTOR kinase inhibitor and dasatinib as a novel therapeutic strategy for vestibular schwannoma SCIENTIFIC REPORTS Sagers, J. E., Beauchamp, R. L., Zhang, Y., Vasilijic, S., Wu, L., DeSouza, P., Seist, R., Zhou, W., Xu, L., Ramesh, V., Stankovic, K. M. 2020; 10 (1): 4211

  Abstract

  Neurofibromatosis type 2 (NF2) is an inherited disorder characterized by bilateral vestibular schwannomas (VS) that arise from neoplastic Schwann cells (SCs). NF2-associated VSs are often accompanied by meningioma (MN), and the majority of NF2 patients show loss of the NF2 tumor suppressor. mTORC1 and mTORC2-specific serum/glucocorticoid-regulated kinase 1 (SGK1) are constitutively activated in MN with loss of NF2. In a recent high-throughput kinome screen in NF2-null human arachnoidal and meningioma cells, we showed activation of EPH RTKs, c-KIT, and SFK members independent of mTORC1/2 activation. Subsequently, we demonstrated in vitro and in vivo efficacy of combination therapy with the dual mTORC1/2 inhibitor AZD2014 and the multi-kinase inhibitor dasatinib. For these reasons, we investigated activated mTORC1/2 and EPH receptor-mediated signaling in sporadic and NF2-associated VS. Using primary human VS cells and a mouse allograft model of schwannoma, we evaluated the dual mTORC1/2 inhibitor AZD2014 and the tyrosine kinase inhibitor dasatinib as monotherapies and in combination. Escalating dose-response experiments on primary VS cells grown from 15 human tumors show that combination therapy with AZD2014 and dasatinib is more effective at reducing metabolic activity than either drug alone and exhibits a therapeutic effect at a physiologically reasonable concentration (~0.1 µM). In vivo, while AZD2014 and dasatinib each inhibit tumor growth alone, the effect of combination therapy exceeds that of either drug. Co-targeting the mTOR and EPH receptor pathways with these or similar compounds may constitute a novel therapeutic strategy for VS, a condition for which there is no FDA-approved pharmacotherapy.

  View details for DOI 10.1038/s41598-020-60156-6

  View details for Web of Science ID 000560099700001

  View details for PubMedID 32144278

  View details for PubMedCentralID PMC7060236

• Cytokine Levels in Inner Ear Fluid of Young and Aged Mice as Molecular Biomarkers of Noise-Induced Hearing Loss FRONTIERS IN NEUROLOGY Landegger, L. D., Vasilijic, S., Fujita, T., Soares, V. Y., Seist, R., Xu, L., Stankovic, K. M. 2019; 10: 977

  Abstract

  Sensorineural hearing loss (SNHL) is the most common sensory deficit worldwide, frequently caused by noise trauma and aging, with inflammation being implicated in both pathologies. Here, we provide the first direct measurements of proinflammatory cytokines in inner ear fluid, perilymph, of adolescent and 2-year-old mice. The perilymph of adolescent mice exposed to the noise intensity resulting in permanent auditory threshold elevations had significantly increased levels of IL-6, TNF-α, and CXCL1 6 h after exposure, with CXCL1 levels being most elevated (19.3 ± 6.2 fold). We next provide the first immunohistochemical localization of CXCL1 in specific cochlear supporting cells, and its presumed receptor, Duffy antigen receptor for chemokines (DARC), in hair cells and spiral ganglion neurons. Our results demonstrate the feasibility of molecular diagnostics of SNHL using only 0.5 μL of perilymph, and motivate future sub-μL based diagnostics of human SNHL based on liquid biopsy of the inner ear to guide therapy, promote hearing protection, and monitor response to treatment.

  View details for DOI 10.3389/fneur.2019.00977

  View details for Web of Science ID 000485185600001

  View details for PubMedID 31632328

  View details for PubMedCentralID PMC6749100

• Nanostructured endodontic materials mixed with different radiocontrast agentsbiocompatibility study JOURNAL OF MATERIALS SCIENCE-MATERIALS IN MEDICINE Cetenovic, B., Colovic, B., Vasilijic, S., Prokic, B., Pasalic, S., Jokanovic, V., Tepavcevic, Z., Markovic, D. 2018; 29 (12): 190

  Abstract

  The aim of this study was to investigate the biocompatibility of nanostructured materials based on highly active calcium silicates mixed with different radiocontrast agents in comparison to MTA+ using in vitro and in vivo model. Morphology of materials' samples was analyzed using SEM while the phase compositions were identified by XRD. pH values of materials' suspensions were conducted by pH-meter. The cytotoxicity of materials' solutions was tested by MTT test (100, 50, 25 and 12.5 mg/ml). LDH and 3H-thymidine assay were utilized for biocompatibility investigations of materials' eluates (24 h, 7 day and 21 day). Eighteen Guinea pigs were used for intramuscular implantation, as teflon tubes with freshly prepared materials were placed into intramuscular pockets. All samples were composed of round and needle-like particles equally distributed with Ca/Si ratio ~2.7 at%, with the presence of hydrated calcium silicate phases. The pH values of ALBO-MPCA1 and ALBO-MPCA2 were high alkaline, while in case of MTA+ they were lower and continuously declined (p < 0.05). Investigated materials didn't exhibit dose-dependent effect on metabolic activity of L929 cells (p > 0.05). Significant differences in the percentage of cytotoxicity between diluted and undiluted extracts between all tested materials after 24 h and 7 day were noticed (p < 0.05). Increase in L929 cells proliferation was noticed in case of undiluted eluates of ALBO-MPCA1 and ALBO-MPCA2 after 7 day (p < 0.05). There were no statistically significant differences in the intensity of inflammatory response between investigated materials and control group after 60 day (p > 0.05). Evaluation of biocompatibility of both ALBO-MPCA1 and ALBO-MPCA2 indicate their potential clinical use.

  View details for DOI 10.1007/s10856-018-6200-z

  View details for Web of Science ID 000452711900004

  View details for PubMedID 30536136

• Computational repositioning and preclinical validation of mifepristone for human vestibular schwannoma SCIENTIFIC REPORTS Sagers, J. E., Brown, A. S., Vasilijic, S., Lewis, R. M., Sahin, M. I., Landegger, L. D., Perlis, R. H., Kohane, I. S., Welling, D., Patel, C. J., Stankovic, K. M. 2018; 8: 5437

  Abstract

  The computational repositioning of existing drugs represents an appealing avenue for identifying effective compounds to treat diseases with no FDA-approved pharmacotherapies. Here we present the largest meta-analysis to date of differential gene expression in human vestibular schwannoma (VS), a debilitating intracranial tumor, and use these data to inform the first application of algorithm-based drug repositioning for this tumor class. We apply an open-source computational drug repositioning platform to gene expression data from 80 patient tumors and identify eight promising FDA-approved drugs with potential for repurposing in VS. Of these eight, mifepristone, a progesterone and glucocorticoid receptor antagonist, consistently and adversely affects the morphology, metabolic activity, and proliferation of primary human VS cells and HEI-193 human schwannoma cells. Mifepristone treatment reduces VS cell viability more significantly than cells derived from patient meningiomas, while healthy human Schwann cells remain unaffected. Our data recommend a Phase II clinical trial of mifepristone in VS.

  View details for DOI 10.1038/s41598-018-23609-7

  View details for Web of Science ID 000428994100030

  View details for PubMedID 29615643

  View details for PubMedCentralID PMC5882888

• Biocompatibility Investigation of New Endodontic Materials Based on Nanosynthesized Calcium Silicates Combined with Different Radiopacifiers JOURNAL OF ENDODONTICS Cetenovic, B., Prokic, B., Vasilijic, S., Dojcinovic, B., Magic, M., Jokanovic, V., Markovic, D. 2017; 43 (3): 425-432

  Abstract

  The aim of this article was to analyze biocompatibility and bioactivity of new endodontic materials on the basis of nanosynthesized calcium silicates (ALBO-MPCA1 and ALBO-MPCA2) combined with different radiopacifiers in comparison with MTA+.Morphology of the samples was studied by scanning electron microscopy, and the pH and ion release analysis were also assessed. Biocompatibility of materials' eluates (24-hour, 7-day, and 21-day) was conducted by using MTT test. Twelve New Zealand white rabbits were used for intraosseous implantation. Four calvarial defects per animal were created and filled with freshly prepared investigated materials.Samples mostly consisted of agglomerates built up from nanoparticles, preferably spherical and rod-like. There was no significant difference among pH values of materials' eluates after 24 hours (P > .05). The amount of calcium and aluminum ion release decreased, whereas the amount of magnesium and bismuth (ALBO-MPCA1, MTA+) and barium (ALBO-MPCA2) increased during 21-day period. The metabolic activity of cells increased after the extraction time, except in case of undiluted elutes of ALBO-MPCA2 and ALBO-MPCA1 (21-day). Histologic analysis of the samples revealed newly formed bone tissue with moderate inflammation for all investigated materials, which subsided during 90-day period to mild. Both MTA+ and ALBO-MPCA1 were in direct contact with the newly formed bone tissue. After 90 days, statistically significant difference in hard tissue formation was observed in comparison of MTA+ and ALBO-MPCA1 with control group (P < .05).Experimental materials ALBO-MPCA1 and ALBO-MPCA2 possess both biocompatibility and bioactivity. Because ALBO-MPCA1 provokes favorable biological response, it is especially good candidate for further clinical investigations.

  View details for DOI 10.1016/j.joen.2016.10.041

  View details for Web of Science ID 000395971900011

  View details for PubMedID 28231981

• Fast dendritic cells matured with Poly (I:C) may acquire tolerogenic properties CYTOTHERAPY Pavlovic, B., Tomic, S., Dokic, J., Vasilijic, S., Vucevic, D., Lukic, J., Gruden-Movsesijan, A., Ilic, N., Markovic, M., Colic, M. 2015; 17 (12): 1763-1776

  Abstract

  Because of the labor-intensive and time-consuming conventional protocols for the generation of dendritic cells (DCs) as the most promising tools for anti-cancer therapy that enable the induction of a T-helper (Th)1-mediated anti-tumor immune response, the use of short-term protocols has been proposed. However, data on the applicability of such protocols in cancer immunotherapy are quite limited.We compared the phenotypic and functional capability of fast DCs (fDCs) differentiated for 24 h and then matured for 48 h with Poly (I:C), a strong Th1-promoting agent, with donor-matched conventional DCs (cDCs) differentiated for 5 days and matured likewise.Of 12 donors tested, we identified seven whose monocytes failed to develop into immunogenic DCs through the use of fDC protocol, on the basis of incomplete downregulation of CD14, low expression of CD1a and macrophage-like morphology. Such fDCs have significantly lower expression of CD83, CD86, CCR7 and CD40, weaker allo-stimulatory Th1- and Th17-polarizing capacity caused by poor production of interleukin (IL)-12p70 and IL-23 and high production of IL-10, and prominent Th2-polarizing capacity, compared with donor-matched cDCs. Furthermore, such fDCs had tolerogenic properties as judged by higher expression of indolamine dioxigenase-3, IDO-1 and IL-1β and induction of a higher percentage of CD4(+)CD25(+)FoxP3(+) T cells. These findings correlated with increased transforming growth factor (TGF)-β production by fDC-primed CD3(+)T cells and their stronger anti-proliferative capacity.We emphasize that although fDCs could probably be applied as an alternative to cDCs for cancer therapy, the fDC protocol should not be applied to donors whose DCs acquire tolerogenic capabilities.

  View details for DOI 10.1016/j.jcyt.2015.08.001

  View details for Web of Science ID 000365246500010

  View details for PubMedID 26455276

• Necrosis and apoptosis in Trichinella spiralis-mediated tumour reduction Central European Journal of Immunology Vasilev, S., Ilic, N., Gruden-Movsesijan, A., Vasilijic, S., Bosic, M., Sofronic-Milosavljevic, L. 2015; 40 (1): 42-53

  Abstract

  It is known that infection with different pathogens, including helminths, can alter the progression of malignant or other diseases. We studied the effect of chronic Trichinella spiralis infection or muscle larvae excretory-secretory (ES L1) antigens on the malignant tumour growth in the mouse melanoma model system in vivo and in vitro. Our results confirmed that chronic infection with T. spiralis possesses the capacity to slow down the progression of tumour growth, resulting in an impressive reduction in tumour size. We found that the phenomenon could, at least partially, be related to a lower level of tumour necrosis compared to necrosis present in control animals with progressive malignancy course. An increased apoptotic potential among the low percentage of cells within the total tumour cell number in vivo was also observed. ES L1 antigen, as a parasitic product that is released during the chronic phase of infection, reduced the survival and slightly, but significantly increased the apoptosis level of melanoma cells in vitro. Our results imply that powerful Trichinella anti-malignance capacity does not rely only on necrosis and apoptosis but other mechanisms through which infection or parasite products manipulate the tumor establishment and expansion should be considered.

  View details for DOI 10.5114/ceji.2015.50832

  View details for Web of Science ID 000353430800008

  View details for PubMedID 26155183

  View details for PubMedCentralID PMC4472539

• Size-Dependent Effects of Gold Nanoparticles Uptake on Maturation and Antitumor Functions of Human Dendritic Cells In Vitro PLOS ONE Tomic, S., Dokic, J., Vasilijic, S., Ogrinc, N., Rudolf, R., Pelicon, P., Vucevic, D., Milosavljevic, P., Jankovic, S., Anzel, I., Rajkovic, J., Rupnik, M., Friedrich, B., Colic, M. 2014; 9 (5): e96584

  Abstract

  Gold nanoparticles (GNPs) are claimed as outstanding biomedical tools for cancer diagnostics and photo-thermal therapy, but without enough evidence on their potentially adverse immunological effects. Using a model of human dendritic cells (DCs), we showed that 10 nm- and 50 nm-sized GNPs (GNP10 and GNP50, respectively) were internalized predominantly via dynamin-dependent mechanisms, and they both impaired LPS-induced maturation and allostimulatory capacity of DCs, although the effect of GNP10 was more prominent. However, GNP10 inhibited LPS-induced production of IL-12p70 by DCs, and potentiated their Th2 polarization capacity, while GNP50 promoted Th17 polarization. Such effects of GNP10 correlated with a stronger inhibition of LPS-induced changes in Ca2+ oscillations, their higher number per DC, and more frequent extra-endosomal localization, as judged by live-cell imaging, proton, and electron microscopy, respectively. Even when released from heat-killed necrotic HEp-2 cells, GNP10 inhibited the necrotic tumor cell-induced maturation and functions of DCs, potentiated their Th2/Th17 polarization capacity, and thus, impaired the DCs' capacity to induce T cell-mediated anti-tumor cytotoxicity in vitro. Therefore, GNP10 could potentially induce more adverse DC-mediated immunological effects, compared to GNP50.

  View details for DOI 10.1371/journal.pone.0096584

  View details for Web of Science ID 000338029800089

  View details for PubMedID 24802102

  View details for PubMedCentralID PMC4011871

• Inverse production of IL-6 and IL-10 by abdominal aortic aneurysm explant tissues in culture CARDIOVASCULAR PATHOLOGY Vucevic, D., Maravic-Stojkovic, V., Vasilijic, S., Borovic-Labudovic, M., Majstorovic, I., Radak, D., Jevtic, M., Milosavljevic, P., Colic, M. 2012; 21 (6): 482-489

  Abstract

  Abdominal aortic aneurysm is considered an atherosclerosis-related disease, but the mechanisms underlying abdominal aortic aneurysm remain poorly defined. Despite the large number of cytokines identified in an aneurysm sample, the relative importance of particular cytokines in aneurysm formation is unknown. We have studied the production of interleukin-6 and interleukin-10 cytokines in plasma and cultures of abdominal aortic aneurysm explant samples obtained from patients subjected to elective surgery and their correlation with cellular composition.Inflammatory cells from the abdominal aortic aneurysm samples were phenotypically characterized using specific monoclonal antibodies (anti-CD3, -CD4, -CD8, -CD19, -CD38, -CD68, -HLA-DR) by means of immunocytochemistry staining. Production of interleukin-6 and interleukin-10 in culture supernatants of abdominal aortic aneurysm explant samples expanded in vitro for 24 h was measured by enzyme-linked immunosorbent assay.We showed that the levels of interleukin-6 and interleukin-10 in supernatants of abdominal aortic aneurysm sample cultures were higher by 73 and 86 times compared to their levels in plasma, respectively. In individual abdominal aortic aneurysm explant cultures, a negative correlation between interleukin-6 and interleukin-10 production was observed. Such inverse correlation was not detected in plasma. Based on these results, we divided abdominal aortic aneurysm into two cytokine-producing groups and showed that the interleukin-6(hi)/interleukin-10(lo) group contained higher percentages of granulocytes, HLA-DR(+), and CD68(+) cells but lower percentages of lymphocytes and plasma cells compared to the interleukin-6(lo)/interleukin-10(hi) group. Exogenously added interleukin-10 suppresses the production of interleukin-6 by abdominal aortic aneurysm explants.These results suggest that interleukin-6 and interleukin-10 may have a different role in the pathogenesis of abdominal aortic aneurysm.

  View details for DOI 10.1016/j.carpath.2012.02.006

  View details for Web of Science ID 000311068000006

  View details for PubMedID 22445549

• Signaling through Toll-like receptor 3 and Dectin-1 potentiates the capability of human monocyte-derived dendritic cells to promote T-helper 1 and T-helper 17 immune responses CYTOTHERAPY Dragicevic, A., Dzopalic, T., Vasilijic, S., Vucevic, D., Tomic, S., Bozic, B., Colic, M. 2012; 14 (5): 598-607

  Abstract

  Recent studies have shown that the ligation of Toll-like receptor 3 (TLR3) or Dectin-1 on human monocyte-derived dendritic cells (MoDC) elicits their maturation, but with a different outcome on immunomodulation. Therefore the aim of this work was to study the response of MoDC to the combined effect of polyinosinic:polycytydilic acid [Poly (I:C)] and curdlan, selective TLR3 and Dectin-1 agonists, respectively.Immature MoDC, generated from human monocytes, were treated with Poly (I:C), curdlan or their combination for 2 days. Phenotypic characteristics of MoDC were determined by flow cytometry, and cytokine production was measured by enzyme-linked immunosorbent assay (ELISA) and FlowCytomix, while the stimulatory capability of MoDC was tested using a mixed leukocyte reaction assay.The combination of Poly (I:C) and curdlan induced phenotypic maturation of MoDC with the capability to stimulate an alloreactive response. Such treated MoDC up-regulated the production of interleukin (IL)-12, IL-23 and IL-10, compared with the effect of Poly (I:C) alone. Curdlan-treated MoDC stimulated the production of IL-17 by alloreactive CD4 (+) T cells more strongly than Poly (I:C)-treated MoDC. The opposite effect was observed for interferon(IFN)-γ production. When combined, these agonists primed MoDC to increase further the production of IFN-γ by CD4 (+) T cells in co-culture, especially those of naive (CD45RA (+)) phenotype, and IL-17 by memory (CD45RO (+)) CD4 (+) T cells.Ligation of TLR3 and Dectin-1 receptor up-regulates T-helper (Th) 1 and Th17 immune responses compared with single agonists. These findings may have therapeutic implications for the use of MoDC in immunotherapy.

  View details for DOI 10.3109/14653249.2012.667873

  View details for Web of Science ID 000302808800009

  View details for PubMedID 22424215

• Immunomodulatory Properties of Mesenchymal Stem Cells Derived from Dental Pulp and Dental Follicle are Susceptible to Activation by Toll-Like Receptor Agonists STEM CELLS AND DEVELOPMENT Tomic, S., Djokic, J., Vasilijic, S., Vucevic, D., Todorovic, V., Supic, G., Colic, M. 2011; 20 (4): 695-708

  Abstract

  Adult mesenchymal stem cells (MSCs) have recently become a potent tool in regenerative medicine. Due to certain shortcomings of obtaining bone marrow MSCs, alternate sources of MSCs have been sought. In this work, we studied MSCs from dental pulp (DP-MSCs) and dental follicle (DF-MSCs), isolated from the same tooth/donor, to define differences in their phenotypic properties, differentiation potential, and immunomodulatory activities. Both cell types showed colony-forming ability and expressed typical MSCs markers, but differed in the levels of their expression. DF-MSCs proliferated faster, contained cells larger in diameter, exhibited a higher potential to form adipocytes and a lower potential to form chondrocytes and osteoblasts, compared with DP-MSCs. In contrast to DF-MSCs, DP-MSCs produced the transforming growth factor (TGF)-β and suppressed proliferation of peripheral blood mononuclear cells, which could be neutralized with anti-TGF-β antibody. The treatment with toll-like receptor 3 (TLR3) agonist augmented the suppressive potential of both cell types and potentiated TGF-β and interleukin-6 secretions by these cells. TLR4 agonist augmented the suppressive potential of DF-MSCs and increased TGF-β production, but abrogated the immunosuppressive activity of DP-MSCs by inhibiting TGF-β production and the expression of indolamine-2,3-dioxygenase-1. Some of these effects correlated with the higher expression of TLR3 and TLR4 by DP-MSCs compared with DF-MSCs. When transplanted in imunocompetent xenogenic host, both cell types induced formation of granulomatous tissue. In conclusion, our results suggest that dental MSCs are functionally different and each of these functions should be further explored in vivo before their specific biomedical applications.

  View details for DOI 10.1089/scd.2010.0145

  View details for Web of Science ID 000289053000013

  View details for PubMedID 20731536

• Differences in T-helper polarizing capability between human monocyte-derived dendritic cells and monocyte-derived Langerhans'-like cells IMMUNOLOGY Rajkovic, I., Dragicevic, A., Vasilijic, S., Bozic, B., Dzopalic, T., Tomic, S., Majstorovic, I., Vucevic, D., Djokic, J., Balint, B., Colic, M. 2011; 132 (2): 217-225

  Abstract

  Langerhans' cells (LCs) represent a specific subset of dendritic cells (DCs) which are important for detecting and processing pathogens that penetrate the skin and epithelial barriers. The aim of our study was to explain what makes their in vitro counterparts - monocyte-derived Langerhans'-like cells (MoLCs) - unique compared with monocyte-derived dendritic cells (MoDCs). Immature MoDCs were generated by incubating peripheral blood monocytes with granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin (IL)-4. The addition of transforming growth factor-β (TGF-β) to this cytokine cocktail resulted in the generation of MoLCs. MoLCs showed a lower expression of CD83, CD86, HLA-DR and CCR7 compared with MoDCs, regardless of their maturational status. Both immature and mature MoLCs secreted higher quantities of IL-23 compared with MoDCs and this finding correlated with a higher secretion of IL-17 in co-culture of MoLCs with allogeneic CD4(+) T cells. Mature MoLCs, which produced higher levels of IL-12 and lower levels of IL-10 compared with mature MoDCs, were more potent at inducing interferon-γ (IFN-γ) production by CD4(+) T cells in the co-culture system. In conclusion, the finding that mature MoLCs stimulate stronger T-helper 1 and T-helper 17 immune responses than mature MoDCs, makes them better candidates for use in the preparation of anti-tumour DC vaccines.

  View details for DOI 10.1111/j.1365-2567.2010.03356.x

  View details for Web of Science ID 000285999600007

  View details for PubMedID 21039466

  View details for PubMedCentralID PMC3050445

• Loxoribine, a selective Toll-like receptor 7 agonist, induces maturation of human monocyte-derived dendritic cells and stimulates their Th-1-and Th-17-polarizing capability INTERNATIONAL IMMUNOPHARMACOLOGY Dzopalic, T., Dragicevic, A., Vasilijic, S., Vucevic, D., Majstorovic, I., Bozic, B., Balint, B., Colic, M. 2010; 10 (11): 1428-1433

  Abstract

  Recently, a guanosine analog, 7-allyl-7,8-dihydro-8-oxo-guanosine (loxoribine), has been identified as a selective Toll-like receptor (TLR)7 agonist. Bearing in mind the controversy regarding the expression of TLR7 by human myeloid dendritic cells (DCs) and its significance for functions of these cells, the goal of this study was to investigate the effect of loxoribine on differentiation, maturation and functions of human monocyte-derived (Mo)DCs. Immature MoDCs were obtained by cultivation of monocytes for 6 days with recombinant granulocyte macrophage-colony stimulating factor (GM-CSF) and interleukin (IL)-4. These cells were stimulated with loxoribine (250 μM) for an additional 48 h. Phenotypic properties of MoDCs were determined by flow cytometry, cytokine production was assayed by ELISA, whereas their allostimulatory capability was tested using a mixed leukocyte reaction. We showed that loxoribine up-regulated the expression of TLR7, CD40, CD54, CD80, CD83 and CCR7 and stimulated the production of IL-12, IL-23, IL-27 and IL-10 by MoDCs, whereas the level of interferon (IFN)-β was not modulated. Allogeneic CD4(+)T cells in co-culture with loxoribine-treated MoDCs proliferated more strongly, at lower DC/CD4(+)T-cell ratio (1:80), and secreted significantly higher levels of IL-17 and IFN-γ compared to the cultures with control MoDCs. The stimulatory effect of loxoribine on T helper (Th)1 polarization capability of MoDCs was further potentiated by ligation of CD40. In conclusion, our results show that loxoribine stimulated differentiation, maturation, allostimulatory as well as Th1 and Th17 polarization capability of human MoDCs and suggests that these effects might be associated with up-regulation of TLR7 expression, but not increased IFN-β production.

  View details for DOI 10.1016/j.intimp.2010.08.010

  View details for Web of Science ID 000284657000015

  View details for PubMedID 20817120

• Production of IL-10 and IL-12 by antigen-presenting cells in periapical lesions JOURNAL OF ORAL PATHOLOGY & MEDICINE Colic, M., Gazivoda, D., Vasilijic, S., Vucevic, D., Lukic, A. 2010; 39 (9): 690-696

  Abstract

  Interferon-γ (IFN-γ) plays an important role in the pathogenesis of periapical lesions. Its expression is up-regulated by interleukin (IL)-12) and down-regulated by IL-10. The aim of this work was to study the cellular source of these cytokines and their mutual interactions in human periapical lesions.Mononuclear cells, macrophages and dendritic cells were isolated from periapical lesions using plastic adherence and osmotic gradients. Cytokines were measured in culture supernatants by a microbeads fluorescence assay. Phenotypic characteristics of cells were studied by immunocytochemistry, whereas allostimulatory activity of antigen-presenting cells was tested using a mixed leukocyte reaction.We observed the positive correlations between the levels of IL-12 and IFN-γ as well as IL-12 and IL-10 in cultures of mononuclear cells. As IL-10 and IL-12 are produced by dendritic cells and activated macrophages, we examined their contribution to the production of these cytokines. Macrophages, CD14(+) adherent cells, produced high levels of IL-10 and very low levels of IL-12. In contrast, non-adherent, strongly HLA-DR(+) dendritic cells, potent stimulators of the alloreactive T-cell response, produced low levels of IL-10 and moderate levels of IL-12. Dendritic cells stimulated the production of IFN-γ by allogeneic CD4(+) T cells. In contrast, the level of IFN-γ was significantly decreased and the production of IL-10 was enhanced by addition of macrophages to the culture system.Our results suggest that a fine balance between the production of IL-10 and IL-12 by different antigen-presenting cells, through IFN-γ, may control the course of chronic inflammation in periapical lesions.

  View details for DOI 10.1111/j.1600-0714.2010.00925.x

  View details for Web of Science ID 000282377600008

  View details for PubMedID 20819132

• Immunomodulatory Activity of IL-27 in Human Periapical Lesions JOURNAL OF DENTAL RESEARCH Colic, M., Gazivoda, D., Majstorovic, I., Dragicevic, A., Vasilijic, S., Rudolf, R., Milosavljevic, P., Vucevic, D. 2009; 88 (12): 1142-1147

  Abstract

  IL-27, a cytokine with pro-inflammatory and anti-inflammatory properties, is a new member of the IL-6/IL-12 family, whose function in periapical lesions is unknown. We hypothesized that the production of IL-27 and its effect depend upon the type of immune/inflammatory response and clinical presentation of periapical lesions. We tested this hypothesis by studying the expression and function of IL-27 in human periapical lesions, both in situ and in culture. Immunohistochemistry demonstrated the strongest expression of IL-27 by endothelial cells and mononuclear phagocytes. Its production by periapical lesion mononuclear cells (PL-MNC), especially in symptomatic lesions, was significantly higher compared with that in peripheral blood MNC and correlated with the frequency of CD14(+) and CD3(+) cells. Exogenous IL-27 stimulated Th1 and down-regulated Th17 cytokine production by PL-MNC from symptomatic lesions, but down-regulated Th1 and Th2 responses in asymptomatic lesions. These findings suggest that IL-27 is an immunomodulatory cytokine in periapical lesions, with complex biological effects.

  View details for DOI 10.1177/0022034509351407

  View details for Web of Science ID 000271993600012

  View details for PubMedID 19897784

• Systemic and intraperitoneal proinflammatory cytokine profiles in patients on continuous ambulatory peritoneal dialysis. Advances in peritoneal dialysis. Conference on Peritoneal Dialysis Maksic, D., Vasilijic, S., Colic, M., Stankovic-Popovic, V., Bokonjic, D. 2009; 25: 50-5

  Abstract

  Our cross-sectional study included 44 patients (27 men, 17 women; mean age: 57.12 +/- 16.66 years; mean dialysis treatment period: 3.59 +/- 2.67 years) on continuous ambulatory peritoneal dialysis (CAPD). Of the 44 patients, 21 were using standard solutions (Stay*Safe, ANDY-disc: Fresenius Medical Care, Bad Homburg, Germany), and 23 were using biocompatible solutions (Gambrosol Trio: Gambro Lundia AB, Lund, Sweden; Stay*Safe Balance: Fresenius Medical Care). In all CAPD patients dialyzed longer than 6 months, we analyzed levels of interleukin 1beta (IL-1beta), tumor necrosis factor alpha (TNFalpha), and interleukin 6 (IL-6) in serum and dialysis effluent when patients were free of acute infection-related (CAPD peritonitis, exit-site infection, other acute infections) complications. In a control group of 20 patients with chronic renal failure [CRF (stages IV and V)], we also determined serum levels of the same cytokines. Levels of the inflammatory cytokines were measured using specific commercial ELISA kits (BioSource, Camarillo, CA, U.S.A.). Statistical analysis of the results was performed using commercial statistics software for the PC (Statistica for Windows, rev. 4.5: StatSoft, Tulsa, OK, U.S.A.). Serum levels of LL-1beta and IL-6 were not statistically significantly different between the patients on CAPD, regardless of the type of dialysis the used, and between the patients and the control group with CRF. Serum levels of TNFalpha, unlike those for IL-1beta and IL-6, were statistically significantly higher in patients on CAPD than in the control group with CRF (13.20 +/- 3.23 pg/mL vs. 5.59 +/- 4.54 pg/mL, p < 0.001, Mann-Whitney test). Serum and effluent IL-1beta levels in patients on CAPD for less than 1 year and more than 1 year did not significantly differ, but effluent IL-6 levels were significantly higher than serum IL-6 levels in both groups of patients, and effluent IL-6 levels were significantly higher in CAPD patients dialyzed for more than 1 year than in patients dialyzed for less than 1 year. Serum and intraperitoneal (IP) levels of the examined cytokines did not significantly differ in patients on standard and biocompatible solutions, but a trend toward lower IP levels of IL-6 was seen in patients on biocompatible solutions. Residual renal function and number of episodes of CAPD peritonitis had no important effect on serum and IP levels of the examined cytokines. Elevated serum levels of TNFalpha and significant local IL-6 production in our CAPD patients indirectly confirmed the importance of peritoneal dialysis (PD) in amplifying the chronic inflammation that substantially depends on duration of dialysis treatment.

  View details for PubMedID 19886317

• Proinflammatory and immunoregulatory mechanisms in periapical lesions MOLECULAR IMMUNOLOGY Colic, M., Gazivoda, D., Vucevic, D., Vasilijic, S., Rudolf, R., Lukic, A. 2009; 47 (1): 101-113

  Abstract

  Proinflammatory and immunoregulatory cytokines are important for the pathogenesis of periapical lesions. However, little is known about how their functions are balanced and controlled at different phases of lesion development. The aim of this study was to examine the relationship between the production of Th1, Th2, Th17 and T regulatory cell (T reg) cytokines by human periapical lesion mononuclear cells (PL-MNC) in culture and their correlation with cellular composition and clinical presentation of the lesions. We show that symptomatic lesions are characterized by the infiltration of neutrophils, high production of IL-17, positive correlation between IL-17 and IFN-gamma, but not between IL-17 and IL-23 production. Most IL-17(+) cells coexpressed IFN-gamma. Asymptomatic lesions were phenotypically heterogeneous. The lesions with the predominance of T cells over B cells/plasma cells expressed higher levels of IFN-gamma which correlated with higher production of IL-12 and the frequency of macrophages. In contrast, in most B-type lesions higher levels of IL-5 and TGF-beta were observed, as well as positive correlation between the production of TGF-beta and IL-10. The addition of Th cytokines in PL-MNC cultures confirmed that Th1, Th2 and Th17 cytokines are mutually antagonistic, except that IL-17, unexpectedly, augmented the production of IFN-gamma. IL-10 and TGF-beta inhibited the production of both Th1 and Th17 cytokines. Dendritic cells (DCs) from periapical lesions, composed of immature (CD83(-)), and mature (CD83(+)) myeloid type DCs and plasmacytoid (BDCA2(+)) DCs produced higher levels of IL-12 and IL-23 but lower levels of IL-10 and TNF-alpha than monocyte (Mo) -derived DCs. IL-23 stimulated the production of IL-17 by PL-MNC, whereas the secretion of IFN-gamma was enhanced by both IL-12 and IL-23. Cumulatively, these results suggest that: (1) Th1 immune response is most probably important for all stages of periapical lesion development; (2) Th2 and immunoregulatory cytokines are more significant for advanced types of lesions with the predominance of B cells/plasma cells; (3) Th17 immune response seems to play a dominant role in exacerbating inflammation.

  View details for DOI 10.1016/j.molimm.2009.01.011

  View details for Web of Science ID 000272366200016

  View details for PubMedID 19232436

• Regulatory T-cells in Periapical Lesions JOURNAL OF DENTAL RESEARCH Colic, M., Gazivoda, D., Vucevic, D., Majstorovic, I., Vasilijic, S., Rudolf, R., Brkic, Z., Milosavljevic, P. 2009; 88 (11): 997-1002

  Abstract

  CD4(+)CD25(hi)Foxp3(+) regulatory T-cells (Tregs) are of crucial importance in regulating the immune response, including the control of any defense against infection. Their presence in periapical lesions has not been demonstrated, as yet. We hypothesized that Tregs infiltrate periapical lesions, where they inhibit T-cell proliferation. The aim of this study was to characterize Tregs in periapical lesions by confocal microscopy, flow cytometry, and functional assays. We showed that CD4(+)CD25(hi)Foxp3(+) cells in periapical lesions expressed IL-10 and TGF-beta. Their frequency was significantly higher than in peripheral blood and correlated with the levels of TGF-beta and IL-10 in culture supernatants of periapical lesion mononuclear cells. Tregs inhibited the proliferation of responder T-cells in vitro, at least in part, by stimulating the production of IL-10. These findings suggest that CD4(+)CD25(hi)Foxp3(+) cells in periapical lesions may play regulatory roles in controlling local immune/inflammatory processes.

  View details for DOI 10.1177/0022034509347090

  View details for Web of Science ID 000270827500004

  View details for PubMedID 19828886

• Interleukin-17 plays a role in exacerbation of inflammation within chronic periapical lesions EUROPEAN JOURNAL OF ORAL SCIENCES Colic, M., Vasilijic, S., Gazivoda, D., Vucevic, D., Marjanovic, M., Lukic, A. 2007; 115 (4): 315-320

  Abstract

  Interleukin (IL)-17 plays an important role in inflammation and certain autoimmune diseases. However, its role in the pathogenesis of chronic dental periapical lesions has not been studied. Periapical lesion mononuclear cells (PL-MNC) were isolated from inflammatory cells and phenotypically analyzed by immunocytochemistry. The cells were cultured in vitro and IL-17 and IL-8 were measured in the culture supernatants. Controls were peripheral blood (PB) MNC. The level of IL-17 and the proportion of neutrophils were significantly higher in symptomatic lesions. In addition, the production of IL-17 was higher in culture supernatants of PL-MNC isolated from lesions with a predominance of T cells, and the IL-17 concentration correlated with the proportion of CD3+ and CD4+ cells. There was a positive correlation between the levels of IL-17 and IL-8 in the group of symptomatic lesions. The relationship between these cytokines was additionally confirmed on the basis of augmented production of IL-8 by both PL-MNC and PB-MNC treated with IL-17. Our results suggest that IL-17, by stimulating the production of IL-8, may play a role in exacerbating inflammation within chronic periapical lesions.

  View details for DOI 10.1111/j.1600-0722.2007.00460.x

  View details for Web of Science ID 000248962900011

  View details for PubMedID 17697172

• Characterization of antigen-presenting cells in human apical periodontitis lesions by flow cytometry and immunocytochemistry INTERNATIONAL ENDODONTIC JOURNAL Lukic, A., Vasilijic, S., Majstorovic, I., Vucevic, D., Mojsilovic, S., Gazivoda, D., Danilovic, V., Petrovic, R., Colic, M. 2006; 39 (8): 626-636

  Abstract

  To analyse phenotypic characteristics of antigen-presenting cells (APC), isolated from human periapical lesions by flow cytometry and immunocytochemistry.Sixteen periapical lesions were digested for 15 min with 0.05% collagenase. Mononuclear cells, separated from other inflammatory cells by density centrifugation, were processed for flow cytometry and/or immunocytochemistry. Single and double immunostainings were performed using monoclonal antibodies specific for human CD45, CD3, CD19, CD14, HLA-DR, CD1a, CD83 and CD123.Antigen-presenting cells (HLA-DR(+) cells) represented 32.9 +/- 17.8% of total mononuclear cells. Amongst them, B cells (HLA-DR(+) CD19(+)) were the predominant APC population, followed by activated macrophages (HLA-DR(+) CD14(+)), dendritic cells (DC) (HLA-DR(+) CD14(-) CD19(-) CD3(-)) and activated T cells (HLA-DR(+) CD3(+)). Based on the predominance of T cells (CD3(+)) or B cells and plasma cells (CD19(+) and CD19(lo), respectively) amongst mononuclear cell infiltrates, lesions were divided into T- and B-types. The percentage of DC in T-type lesions (27.1 +/- 6.8% of total HLA-DR(+) cells) was higher, compared with B-type lesions (10.3 +/- 5.2%) (P < 0.01). Within the DC population, the percentages of CD1a (Langerhans cell type) and CD123 (probably plasmacytoid DC type) did not differ significantly between the groups (P > 0.05). However, the percentage of mature DC (CD83(+)) was significantly higher in T-type periapical lesions (P < 0.05).Flow cytometry and immunocytochemistry are suitable methods for phenotypic analysis of APC after their isolation from human periapical lesions. APC, that were phenotypically heterogeneous, constituted a significant component of infiltrating cells. Lesions with the predominance of T cells were characterized by a higher proportion of mature DC (HLA-DR(+)CD83(+) cells) than lesions with predominance of B cells/plasma cells.

  View details for DOI 10.1111/j.1365-2591.2006.01125.x

  View details for Web of Science ID 000238887500004

  View details for PubMedID 16872457

• Dendritic cells acquire tolerogenic properties at the site of sterile granulomatous inflammation Cellular Immunology Vasilijic, S., Savic, D., Vasilev, S., Vucevic, D., Gasic, S., Majstorovic, I., Jankovic, S., Colic, M. 2005; 233 (2)
• Granulocyte-macrophage colony stimulating factor is an antiapoptotic cytokine for thymic dendritic cells and a significant modulator of their accessory function IMMUNOLOGY LETTERS Vasilijic, S., Colic, M., Vucevic, D. 2003; 86 (1): 99-112

  Abstract

  Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a growth-promoting factor for myeloid-derived dendritic cells (DC) but not for lymphoid DC. The data about its effect on thymic DC (TDC), which are both of lymphoid and myeloid origin, are very scarce. Using an in vitro model, we demonstrated in this work that GM-CSF significantly increased the survival of rat TDC in culture by inhibiting their apoptosis and the effect correlated with up-regulation of Bcl-2 expression. GM-CSF also stimulated differentiation and maturation of TDC as judged by higher expression of MHC class I and II molecules, CD54, CD80 and CD86. These changes correlated with stronger stimulatory activity of GM-CSF-pulsed TDC in syngeneic thymocyte proliferation assay and MLR. The stimulatory potential of TDC was further increased when thymocytes were cultivated with an anti-alphabeta TCR (R73) monoclonal antibody (mAb). The influence of unstimulated TDC on proliferation of thymocytes was inhibited by anti-CD86 but not anti-CD80 mAb, whereas in cultures with GM-CSF-treated TDC both mAbs exerted an additive blocking effect. After separation of TDC on CD11b(+) and CD11b(-) we demonstrated that GM-CSF inhibited apoptosis and potentiated accessory activity of both TDC subsets independently of the myeloid marker expression. Cummulatively, our results suggest that GM-CSF is one of the regulatory cytokine involved in survival, maturation, differentiation and accessory function of TDC.

  View details for DOI 10.1016/S0165-2478(02)00295-X

  View details for Web of Science ID 000181571200014

  View details for PubMedID 12600752