Yueh-hsiu Chien

All Publications

• γδ T cell antigen receptor polyspecificity enables T cell responses to a broad range of immune challenges. Proceedings of the National Academy of Sciences of the United States of America Guo, J., Chowdhury, R. R., Mallajosyula, V., Xie, J., Dubey, M., Liu, Y., Li, J., Wei, Y. L., Palanski, B. A., Wang, C., Qiu, L., Ohanyan, M., Kask, O., Sola, E., Kamalyan, L., Lewis, D. B., Scriba, T. J., Davis, M. M., Dodd, D., Zeng, X., Chien, Y. H. 2024; 121 (4): e2315592121

  Abstract

  γδ T cells are essential for immune defense and modulating physiological processes. While they have the potential to recognize large numbers of antigens through somatic gene rearrangement, the antigens which trigger most γδ T cell response remain unidentified, and the role of antigen recognition in γδ T cell function is contentious. Here, we show that some γδ T cell receptors (TCRs) exhibit polyspecificity, recognizing multiple ligands of diverse molecular nature. These ligands include haptens, metabolites, neurotransmitters, posttranslational modifications, as well as peptides and proteins of microbial and host origin. Polyspecific γδ T cells are enriched among activated cells in naive mice and the responding population in infection. They express diverse TCR sequences, have different functional potentials, and include the innate-like γδ T cells, such as the major IL-17 responders in various pathological/physiological conditions. We demonstrate that encountering their antigenic microbiome metabolite maintains their homeostasis and functional response, indicating that their ability to recognize multiple ligands is essential for their function. Human γδ T cells with similar polyspecificity also respond to various immune challenges. This study demonstrates that polyspecificity is a prevalent feature of γδ T cell antigen recognition, which enables rapid and robust T cell responses to a wide range of challenges, highlighting a unique function of γδ T cells.

  View details for DOI 10.1073/pnas.2315592121

  View details for PubMedID 38227652

• NK-like CD8+ γδ T cells are expanded in persistent Mycobacterium tuberculosis infection. Science immunology Roy Chowdhury, R., Valainis, J. R., Dubey, M., von Boehmer, L., Sola, E., Wilhelmy, J., Guo, J., Kask, O., Ohanyan, M., Sun, M., Huang, H., Huang, X., Nguyen, P. K., Scriba, T. J., Davis, M. M., Bendall, S. C., Chien, Y. H. 2023; 8 (81): eade3525

  Abstract

  The response of gamma delta (γδ) T cells in the acute versus chronic phases of the same infection is unclear. How γδ T cells function in acute Mycobacterium tuberculosis (Mtb) infection is well characterized, but their response during persistent Mtb infection is not well understood, even though most infections with Mtb manifest as a chronic, clinically asymptomatic state. Here, we analyze peripheral blood γδ T cells from a South African adolescent cohort and show that a unique CD8+ γδ T cell subset with features of "memory inflation" expands in chronic Mtb infection. These cells are hyporesponsive to T cell receptor (TCR)-mediated signaling but, like NK cells, can mount robust CD16-mediated cytotoxic responses. These CD8+ γδ T cells comprise a highly focused TCR repertoire, with clonotypes that are Mycobacterium specific but not phosphoantigen reactive. Using multiparametric single-cell pseudo-time trajectory analysis, we identified the differentiation paths that these CD8+ γδ T cells follow to develop into effectors in this infection state. Last, we found that circulating CD8+ γδ T cells also expand in other chronic inflammatory conditions, including cardiovascular disease and cancer, suggesting that persistent antigenic exposure may drive similar γδ T cell effector programs and differentiation fates.

  View details for DOI 10.1126/sciimmunol.ade3525

  View details for PubMedID 37000856

• A TLR7-nanoparticle adjuvant promotes a broad immune response against heterologous strains of influenza and SARS-CoV-2. Nature materials Yin, Q., Luo, W., Mallajosyula, V., Bo, Y., Guo, J., Xie, J., Sun, M., Verma, R., Li, C., Constantz, C. M., Wagar, L. E., Li, J., Sola, E., Gupta, N., Wang, C., Kask, O., Chen, X., Yuan, X., Wu, N. C., Rao, J., Chien, Y., Cheng, J., Pulendran, B., Davis, M. M. 2023

  Abstract

  The ideal vaccine against viruses such as influenza and SARS-CoV-2 must provide a robust, durable and broad immune protection against multiple viral variants. However, antibody responses to current vaccines often lack robust cross-reactivity. Here we describe a polymeric Toll-like receptor 7 agonist nanoparticle (TLR7-NP) adjuvant, which enhances lymph node targeting, and leads to persistent activation of immune cells and broad immune responses. When mixed with alum-adsorbed antigens, this TLR7-NP adjuvant elicits cross-reactive antibodies for both dominant and subdominant epitopes and antigen-specific CD8+ T-cell responses in mice. This TLR7-NP-adjuvanted influenza subunit vaccine successfully protects mice against viral challenge of a different strain. This strategy also enhances the antibody response to a SARS-CoV-2 subunit vaccine against multiple viral variants that have emerged. Moreover, this TLR7-NP augments antigen-specific responses in human tonsil organoids. Overall, we describe a nanoparticle adjuvant to improve immune responses to viral antigens, with promising implications for developing broadly protective vaccines.

  View details for DOI 10.1038/s41563-022-01464-2

  View details for PubMedID 36717665

• Human Coronary Plaque T Cells Are Clonal and Cross-React to Virus and Self. Circulation research Roy Chowdhury, R., D'Addabbo, J., Huang, X., Veizades, S., Sasagawa, K., Louis, D. M., Cheng, P., Sokol, J., Jensen, A., Tso, A., Shankar, V., Wendel, B. S., Bakerman, I., Liang, G., Koyano, T., Fong, R., Nau, A., Ahmad, H., Gopakumar, J. K., Wirka, R., Lee, A., Boyd, J., Woo, Y. J., Quertermous, T., Gulati, G., Jaiswal, S., Chien, Y. H., Chan, C., Davis, M. M., Nguyen, P. K. 2022: 101161CIRCRESAHA121320090

  Abstract

  Once considered primarily a disorder of lipid deposition, coronary artery disease is an incurable, life-threatening disease that is now also characterized by chronic inflammation notable for the buildup of atherosclerotic plaques containing immune cells in various states of activation and differentiation. Understanding how these immune cells contribute to disease progression may lead to the development of novel therapeutic strategies.We used single-cell technology and in vitro assays to interrogate the immune microenvironment of human coronary atherosclerotic plaque at different stages of maturity.In addition to macrophages, we found a high proportion of αβ T cells in the coronary plaques. Most of these T cells lack high expression of CCR7 and L-selectin, indicating that they are primarily antigen-experienced, memory cells. Notably, nearly one-third of these cells express the HLA-DRA surface marker, signifying activation through their TCRs (T-cell receptors). Consistent with this, TCR repertoire analysis confirmed the presence of activated αβ T cells (CD4

  View details for DOI 10.1161/CIRCRESAHA.121.320090

  View details for PubMedID 35430876

• KIR+CD8+ T cells suppress pathogenic T cells and are active in autoimmune diseases and COVID-19. Science (New York, N.Y.) Li, J., Zaslavsky, M., Su, Y., Guo, J., Sikora, M. J., van Unen, V., Christophersen, A., Chiou, S., Chen, L., Li, J., Ji, X., Wilhelmy, J., McSween, A. M., Palanski, B. A., Mallajosyula, V. V., Bracey, N. A., Dhondalay, G. K., Bhamidipati, K., Pai, J., Kipp, L. B., Dunn, J. E., Hauser, S. L., Oksenberg, J. R., Satpathy, A. T., Robinson, W. H., Dekker, C. L., Steinmetz, L. M., Khosla, C., Utz, P. J., Sollid, L. M., Chien, Y., Heath, J. R., Fernandez-Becker, N. Q., Nadeau, K. C., Saligrama, N., Davis, M. M. 2022: eabi9591

  Abstract

  Here we find that CD8+ T cells expressing inhibitory killer cell immunoglobulin-like receptors (KIRs) are the human equivalent of Ly49+CD8+ regulatory T cells in mice and are increased in the blood and inflamed tissues of patients with a variety of autoimmune diseases. Moreover, these CD8+ T cells efficiently eliminated pathogenic gliadin-specific CD4+ T cells from celiac disease patients' leukocytes in vitro. We also find elevated levels of KIR+CD8+ T cells, but not CD4+ regulatory T cells, in COVID-19 patients, which correlated with disease severity and vasculitis. Selective ablation of Ly49+CD8+ T cells in virus-infected mice led to autoimmunity post infection. Our results indicate that in both species, these regulatory CD8+ T cells act uniquely to suppress pathogenic T cells in autoimmune and infectious diseases.

  View details for DOI 10.1126/science.abi9591

  View details for PubMedID 35258337

• Nanoparticle-enabled innate immune stimulation activates endogenous tumor-infiltrating T cells with broad antigen specificities. Proceedings of the National Academy of Sciences of the United States of America Yin, Q., Yu, W., Grzeskowiak, C. L., Li, J., Huang, H., Guo, J., Chen, L., Wang, F., Zhao, F., von Boehmer, L., Metzner, T. J., Leppert, J. T., Chien, Y., Kuo, C. J., Davis, M. M. 2021; 118 (21)

  Abstract

  Tumors are often infiltrated by T lymphocytes recognizing either self- or mutated antigens but are generally inactive, although they often show signs of prior clonal expansion. Hypothesizing that this may be due to peripheral tolerance, we formulated nanoparticles containing innate immune stimulants that we found were sufficient to activate self-specific CD8+ T cells and injected them into two different mouse tumor models, B16F10 and MC38. These nanoparticles robustly activated and/or expanded antigen-specific CD8+ tumor-infiltrating T cells, along with a decrease in regulatory CD4+ T cells and an increase in Interleukin-17 producers, resulting in significant tumor growth retardation or elimination and the establishment of immune memory in surviving mice. Furthermore, nanoparticles with modification of stimulating human T cells enabled the robust activation of endogenous T cells in patient-derived tumor organoids. These results indicate that breaking peripheral tolerance without regard to the antigen specificity creates a promising pathway for cancer immunotherapy.

  View details for DOI 10.1073/pnas.2016168118

  View details for PubMedID 34021082

• Select sequencing of clonally expanded CD8+ T cells reveals limits to clonal expansion. Proceedings of the National Academy of Sciences of the United States of America Huang, H., Sikora, M. J., Islam, S., Chowdhury, R. R., Chien, Y., Scriba, T. J., Davis, M. M., Steinmetz, L. M. 2019

  Abstract

  To permit the recognition of antigens, T cells generate a vast diversity of T cell receptor (TCR) sequences. Upon binding of the TCR to an antigen-MHC complex, T cells clonally expand to establish an immune response. To study antigen-specific T cell clonality, we have developed a method that allows selection of rare cells, based on RNA expression, before in-depth scRNA-seq (named SELECT-seq). We applied SELECT-seq to collect both TCR sequences and then transcriptomes from single cells of peripheral blood lymphocytes activated by a Mycobacterium tuberculosis (Mtb) lysate. TCR sequence analysis allowed us to preferentially select expanded conventional CD8+ T cells as well as invariant natural killer T (iNKT) cells and mucosal-associated invariant T (MAIT) cells. The iNKT and MAIT cells have a highly similar transcriptional pattern, indicating that they carry out similar immunological functions and differ considerably from conventional CD8+ T cells. While there is no relationship between expression profiles and clonal expansion in iNKT or MAIT cells, highly expanded conventional CD8+ T cells down-regulate the interleukin 2 (IL-2) receptor alpha (IL2RA, or CD25) protein and show signs of senescence. This suggests inherent limits to clonal expansion that act to diversify the T cell response repertoire.

  View details for PubMedID 30992377

• Casting a wider net: Immunosurveillance by nonclassical MHC molecules. PLoS pathogens D'Souza, M. P., Adams, E., Altman, J. D., Birnbaum, M. E., Boggiano, C., Casorati, G., Chien, Y., Conley, A., Eckle, S. B., Fruh, K., Gondre-Lewis, T., Hassan, N., Huang, H., Jayashankar, L., Kasmar, A. G., Kunwar, N., Lavelle, J., Lewinsohn, D. M., Moody, B., Picker, L., Ramachandra, L., Shastri, N., Parham, P., McMichael, A. J., Yewdell, J. W. 2019; 15 (2): e1007567

  Abstract

  Most studies of T lymphocytes focus on recognition of classical major histocompatibility complex (MHC) class I or II molecules presenting oligopeptides, yet there are numerous variations and exceptions of biological significance based on recognition of a wide variety of nonclassical MHC molecules. These include alphabeta and gammadelta T cells that recognize different class Ib molecules (CD1, MR-1, HLA-E, G, F, et al.) that are nearly monomorphic within a given species. Collectively, these T cells can be considered "unconventional," in part because they recognize lipids, metabolites, and modified peptides. Unlike classical MHC-specific cells, unconventional T cells generally exhibit limited T-cell antigen receptor (TCR) repertoires and often produce innate immune cell-like rapid effector responses. Exploiting this system in new generation vaccines for human immunodeficiency virus (HIV), tuberculosis (TB), other infectious agents, and cancer was the focus of a recent workshop, "Immune Surveillance by Non-classical MHC Molecules: Improving Diversity for Antigens," sponsored by the National Institute of Allergy and Infectious Diseases. Here, we summarize salient points presented regarding the basic immunobiology of unconventional T cells, recent advances in methodologies to measure unconventional T-cell activity in diseases, and approaches to harness their considerable clinical potential.

  View details for PubMedID 30789961

• Opposing T cell responses in experimental autoimmune encephalomyelitis. Nature Saligrama, N. n., Zhao, F. n., Sikora, M. J., Serratelli, W. S., Fernandes, R. A., Louis, D. M., Yao, W. n., Ji, X. n., Idoyaga, J. n., Mahajan, V. B., Steinmetz, L. M., Chien, Y. H., Hauser, S. L., Oksenberg, J. R., Garcia, K. C., Davis, M. M. 2019

  Abstract

  Experimental autoimmune encephalomyelitis is a model for multiple sclerosis. Here we show that induction generates successive waves of clonally expanded CD4+, CD8+ and γδ+ T cells in the blood and central nervous system, similar to gluten-challenge studies of patients with coeliac disease. We also find major expansions of CD8+ T cells in patients with multiple sclerosis. In autoimmune encephalomyelitis, we find that most expanded CD4+ T cells are specific for the inducing myelin peptide MOG35-55. By contrast, surrogate peptides derived from a yeast peptide major histocompatibility complex library of some of the clonally expanded CD8+ T cells inhibit disease by suppressing the proliferation of MOG-specific CD4+ T cells. These results suggest that the induction of autoreactive CD4+ T cells triggers an opposing mobilization of regulatory CD8+ T cells.

  View details for DOI 10.1038/s41586-019-1467-x

  View details for PubMedID 31391585

• Author Correction: A multi-cohort study of the immune factors associated with M. tuberculosis infection outcomes. Nature Roy Chowdhury, R., Vallania, F., Yang, Q., Lopez Angel, C. J., Darboe, F., Penn-Nicholson, A., Rozot, V., Nemes, E., Malherbe, S. T., Ronacher, K., Walzl, G., Hanekom, W., Davis, M. M., Winter, J., Chen, X., Scriba, T. J., Khatri, P., Chien, Y. 2018

  Abstract

  The spelling of author Qianting Yang was corrected; the affiliation of author Stephanus T. Malherbe was corrected; and graphs in Fig. 4b and c were corrected owing to reanalysis of the data into the correct timed intervals.

  View details for PubMedID 30377311

• A multi-cohort study of the immune factors associated with M. tuberculosis infection outcomes NATURE Chowdhury, R., Vallania, F., Yang, Q., Angel, C., Darboe, F., Penn-Nicholson, A., Rozot, V., Nemes, E., Malherbe, S. T., Ronacher, K., Walzl, G., Hanekom, W., Davis, M. M., Winter, J., Chen, X., Scriba, T. J., Khatri, P., Chien, Y. 2018; 560 (7720): 644-+

  View details for DOI 10.1038/s41586-018-0439-x

  View details for Web of Science ID 000443218600050

• A multi-cohort study of the immune factors associated with M. tuberculosis infection outcomes. Nature Roy Chowdhury, R., Vallania, F., Yang, Q., Lopez Angel, C. J., Darboe, F., Penn-Nicholson, A., Rozot, V., Nemes, E., Malherbe, S. T., Ronacher, K., Walzl, G., Hanekom, W., Davis, M. M., Winter, J., Chen, X., Scriba, T. J., Khatri, P., Chien, Y. 2018

  Abstract

  Most infections with Mycobacterium tuberculosis (Mtb) manifest as a clinically asymptomatic, contained state, known as latent tuberculosis infection, that affects approximately one-quarter of the global population1. Although fewer than one in ten individuals eventually progress to active disease2, tuberculosis is a leading cause of death from infectious disease worldwide3. Despite intense efforts, immune factors that influence the infection outcomes remain poorly defined. Here we used integrated analyses of multiple cohorts to identify stage-specific host responses to Mtb infection. First, using high-dimensional mass cytometry analyses and functional assays of a cohort of South African adolescents, we show that latent tuberculosis is associated with enhanced cytotoxic responses, which are mostly mediated by CD16 (also known as FcgammaRIIIa) and natural killer cells, and continuous inflammation coupled with immune deviations in both T and B cell compartments.Next, using cell-type deconvolution of transcriptomic data from several cohorts of different ages, genetic backgrounds, geographical locations and infection stages, we show that although deviations in peripheral B and T cell compartments generally start at latency, they are heterogeneous across cohorts. However, an increase in the abundance of circulating natural killer cells in tuberculosis latency, with a corresponding decrease during active disease and a return to baseline levels upon clinical cure are features that are common to all cohorts. Furthermore, by analysing three longitudinal cohorts, we find that changes inperipheral levels of natural killer cells can inform disease progression and treatment responses, and inversely correlate with the inflammatory state of the lungs of patients with active tuberculosis. Together, our findings offer crucial insights into the underlying pathophysiology of tuberculosis latency, and identify factors that may influence infection outcomes.

  View details for PubMedID 30135583

• Novel M-CSF-producing gamma delta T cells protect against recurrent malaria Mamedov, M. R., Scholzen, A., Nair, R. V., Cumnock, K., Kenkel, J. A., Oliveira, J. M., Trujillo, D. L., Saligrama, N., Zhang, Y., Rubelt, F., Schneider, D. S., Chien, Y., Sauerwein, R., Davis, M. M. AMER ASSOC IMMUNOLOGISTS. 2018

  View details for Web of Science ID 000459977700312

• Does selecting ligand shape gamma delta-TCR repertoire? PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA Chien, Y. 2018; 115 (16): E3606

  View details for PubMedID 29615511

• From T Cell Receptor to Antigen, Systems Approach to Discovering T Cell Antigen(s) in Multiple Sclerosis and Experimental Autoimmune Encephalomyelitis Saligrama, N., Zhao, F., Fernandes, R. A., Serratelli, W. S., Louis, D. M., Chien, Y., Garcia, C. K., Oksenberg, J., Davis, M. M. SAGE PUBLICATIONS LTD. 2018: 5

  View details for Web of Science ID 000429034600007

• A Macrophage Colony-Stimulating-Factor-Producing γδ T Cell Subset Prevents Malarial Parasitemic Recurrence. Immunity Mamedov, M. R., Scholzen, A. n., Nair, R. V., Cumnock, K. n., Kenkel, J. A., Oliveira, J. H., Trujillo, D. L., Saligrama, N. n., Zhang, Y. n., Rubelt, F. n., Schneider, D. S., Chien, Y. H., Sauerwein, R. W., Davis, M. M. 2018; 48 (2): 350–63.e7

  Abstract

  Despite evidence that γδ T cells play an important role during malaria, their precise role remains unclear. During murine malaria induced by Plasmodium chabaudi infection and in human P. falciparum infection, we found that γδ T cells expanded rapidly after resolution of acute parasitemia, in contrast to αβ T cells that expanded at the acute stage and then declined. Single-cell sequencing showed that TRAV15N-1 (Vδ6.3) γδ T cells were clonally expanded in mice and had convergent complementarity-determining region 3 sequences. These γδ T cells expressed specific cytokines, M-CSF, CCL5, CCL3, which are known to act on myeloid cells, indicating that this γδ T cell subset might have distinct functions. Both γδ T cells and M-CSF were necessary for preventing parasitemic recurrence. These findings point to an M-CSF-producing γδ T cell subset that fulfills a specialized protective role in the later stage of malaria infection when αβ T cells have declined.

  View details for PubMedID 29426701

• Concerted T cell response in experimental autoimmune encephalomyelitis and multiple sclerosis Saligrama, N., Zhao, F., Fernandes, R., Serratelli, W. S., Louis, D. M., Chien, Y. C., Garcia, C. K., Oksenberg, J., Davis, M. M. SAGE PUBLICATIONS LTD. 2017: 201

  View details for Web of Science ID 000413730201026

• DCs stemming from microbiota adaptation specifically drive ?dT17 proliferation and activation. Microbiome Fleming, C., Cai, Y., Sun, X., Jala, V. R., Xue, F., Morrissey, S., Wei, Y., Chien, Y., Zhang, H., Haribabu, B., Huang, J., Yan, J. 2017; 5 (1): 46-?

  Abstract

  IL-17-producing γδT cells (γδT17) promote autoinflammatory diseases and cancers. Yet, γδT17 peripheral regulation has not been thoroughly explored especially in the context of microbiota-host interaction. The potent antigen-presenting CD103(+) dendritic cell (DC) is a key immune player in close contact with both γδT17 cells and microbiota. This study presents a novel cellular network among microbiota, CD103(+) DCs, and γδT17 cells.Immunophenotyping of IL-17r(-/-) mice and IL-17r(-/-) IRF8(-/-) mice were performed by ex vivo immunostaining and flow cytometric analysis. We observed striking microbiome differences in the oral cavity and gut of IL-17r(-/-) mice by sequencing 16S rRNA gene (v1-v3 region) and analyzed using QIIME 1.9.0 software platform. Principal coordinate analysis of unweighted UniFrac distance matrix showed differential clustering for WT and IL-17r(-/-) mice.We found drastic homeostatic expansion of γδT17 in all major tissues, most prominently in cervical lymph nodes (cLNs) with monoclonal expansion of Vγ6 γδT17 in IL-17r(-/-) mice. Ki-67 staining and in vitro CFSE assays showed cellular proliferation due to cell-to-cell contact stimulation with microbiota-activated CD103(+) DCs. A newly developed double knockout mice model for IL-17r and CD103(+) DCs (IL-17r(-/-)IRF8(-/-)) showed a specific reduction in Vγ6 γδT17. Vγ6 γδT17 expansion is inhibited in germ-free mice and antibiotic-treated specific pathogen-free (SPF) mice. Microbiota transfer using cohousing of IL-17r(-/-) mice with wildtype mice induces γδT17 expansion in the wildtype mice with increased activated CD103(+) DCs in cLNs. However, microbiota transfer using fecal transplant through oral gavage to bypass the oral cavity showed no difference in colon or systemic γδT17 expansion.These findings reveal for the first time that γδT17 cells are regulated by microbiota dysbiosis through cell-to-cell contact with activated CD103(+) DCs leading to drastic systemic, monoclonal expansion. Microbiota dysbiosis, as indicated by drastic bacterial population changes at the phylum and genus levels especially in the oral cavity, was discovered in mice lacking IL-17r. This network could be very important in regulating both microbiota and immune players. This critical regulatory pathway for γδT17 could play a major role in IL-17-driven inflammatory diseases and needs further investigation to determine specific targets for future therapeutic intervention.

  View details for DOI 10.1186/s40168-017-0263-9

  View details for PubMedID 28438184

• Microbiota-activated CD103(+) DCs stemming from microbiota adaptation specifically drive gamma delta T17 proliferation and activation MICROBIOME Fleming, C., Cai, Y., Sun, X., Jala, V. R., Xue, F., Morrissey, S., Wei, Y., Chien, Y., Zhang, H., Haribabu, B., Huang, J., Yan, J. 2017; 5

  Abstract

  IL-17-producing γδT cells (γδT17) promote autoinflammatory diseases and cancers. Yet, γδT17 peripheral regulation has not been thoroughly explored especially in the context of microbiota-host interaction. The potent antigen-presenting CD103(+) dendritic cell (DC) is a key immune player in close contact with both γδT17 cells and microbiota. This study presents a novel cellular network among microbiota, CD103(+) DCs, and γδT17 cells.Immunophenotyping of IL-17r(-/-) mice and IL-17r(-/-) IRF8(-/-) mice were performed by ex vivo immunostaining and flow cytometric analysis. We observed striking microbiome differences in the oral cavity and gut of IL-17r(-/-) mice by sequencing 16S rRNA gene (v1-v3 region) and analyzed using QIIME 1.9.0 software platform. Principal coordinate analysis of unweighted UniFrac distance matrix showed differential clustering for WT and IL-17r(-/-) mice.We found drastic homeostatic expansion of γδT17 in all major tissues, most prominently in cervical lymph nodes (cLNs) with monoclonal expansion of Vγ6 γδT17 in IL-17r(-/-) mice. Ki-67 staining and in vitro CFSE assays showed cellular proliferation due to cell-to-cell contact stimulation with microbiota-activated CD103(+) DCs. A newly developed double knockout mice model for IL-17r and CD103(+) DCs (IL-17r(-/-)IRF8(-/-)) showed a specific reduction in Vγ6 γδT17. Vγ6 γδT17 expansion is inhibited in germ-free mice and antibiotic-treated specific pathogen-free (SPF) mice. Microbiota transfer using cohousing of IL-17r(-/-) mice with wildtype mice induces γδT17 expansion in the wildtype mice with increased activated CD103(+) DCs in cLNs. However, microbiota transfer using fecal transplant through oral gavage to bypass the oral cavity showed no difference in colon or systemic γδT17 expansion.These findings reveal for the first time that γδT17 cells are regulated by microbiota dysbiosis through cell-to-cell contact with activated CD103(+) DCs leading to drastic systemic, monoclonal expansion. Microbiota dysbiosis, as indicated by drastic bacterial population changes at the phylum and genus levels especially in the oral cavity, was discovered in mice lacking IL-17r. This network could be very important in regulating both microbiota and immune players. This critical regulatory pathway for γδT17 could play a major role in IL-17-driven inflammatory diseases and needs further investigation to determine specific targets for future therapeutic intervention.

  View details for DOI 10.1186/s40168-017-0263-9

  View details for Web of Science ID 000399873100002

  View details for PubMedCentralID PMC5404689

• Defective Signaling in the JAK-STAT Pathway Tracks with Chronic Inflammation and Cardiovascular Risk in Aging Humans. Cell systems Shen-Orr, S. S., Furman, D., Kidd, B. A., Hadad, F., Lovelace, P., Huang, Y., Rosenberg-Hasson, Y., Mackey, S., Grisar, F. A., Pickman, Y., Maecker, H. T., Chien, Y., Dekker, C. L., Wu, J. C., Butte, A. J., Davis, M. M. 2016; 3 (4): 374-384 e4

  Abstract

  Chronic inflammation, a decline in immune responsiveness, and reduced cardiovascular function are all associated with aging, but the relationships among these phenomena remain unclear. Here, we longitudinally profiled a total of 84 signaling conditions in 91 young and older adults and observed an age-related reduction in cytokine responsiveness within four immune cell lineages, most prominently T cells. The phenotype can be partially explained by elevated baseline levels of phosphorylated STAT (pSTAT) proteins and a different response capacity of naive versus memory T cell subsets to interleukin 6 (IL-6), interferon α (IFN-α), and, to a lesser extent, IL-21 and IFN-γ. Baseline pSTAT levels tracked with circulating levels of C-reactive protein (CRP), and we derived a cytokine response score that negatively correlates with measures of cardiovascular disease, specifically diastolic dysfunction and atherosclerotic burden, outperforming CRP. Thus, we identified an immunological link between inflammation, decreased cell responsiveness in the JAK-STAT pathway, and cardiovascular aging. Targeting chronic inflammation may ameliorate this deficiency in cellular responsiveness and improve cardiovascular function.

  View details for DOI 10.1016/j.cels.2016.09.009

  View details for PubMedID 27746093

• Changes in Brain Activation Associated with Spontaneous Improvization and Figural Creativity After Design-Thinking-Based Training: A Longitudinal fMRI Study. Cerebral cortex Saggar, M., Quintin, E., Bott, N. T., Kienitz, E., Chien, Y., Hong, D. W., Liu, N., Royalty, A., Hawthorne, G., Reiss, A. L. 2016

  Abstract

  Creativity is widely recognized as an essential skill for entrepreneurial success and adaptation to daily-life demands. However, we know little about the neural changes associated with creative capacity enhancement. For the first time, using a prospective, randomized control design, we examined longitudinal changes in brain activity associated with participating in a five-week design-thinking-based Creative Capacity Building Program (CCBP), when compared with Language Capacity Building Program (LCBP). Creativity, an elusive and multifaceted construct, is loosely defined as an ability to produce useful/appropriate and novel outcomes. Here, we focus on one of the facets of creative thinking-spontaneous improvization. Participants were assessed pre- and post-intervention for spontaneous improvization skills using a game-like figural Pictionary-based fMRI task. Whole-brain group-by-time interaction revealed reduced task-related activity in CCBP participants (compared with LCBP participants) after training in the right dorsolateral prefrontal cortex, anterior/paracingulate gyrus, supplementary motor area, and parietal regions. Further, greater cerebellar-cerebral connectivity was observed in CCBP participants at post-intervention when compared with LCBP participants. In sum, our results suggest that improvization-based creative capacity enhancement is associated with reduced engagement of executive functioning regions and increased involvement of spontaneous implicit processing.

  View details for PubMedID 27307467

• Detection, phenotyping, and quantification of antigen-specific T cells using a peptide-MHC dodecamer PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA Huang, J., Zeng, X., Sigal, N., Lund, P. J., Su, L. F., Huang, H., Chien, Y., Davis, M. M. 2016; 113 (13): E1890-E1897

  Abstract

  Here we report a peptide-MHC (pMHC) dodecamer as a "next generation" technology that is a significantly more sensitive and versatile alternative to pMHC tetramers for the detection, isolation, and phenotypic analysis of antigen-specific T cells. In particular, dodecamers are able to detect two- to fivefold more antigen-specific T cells in both human and murine CD4(+)and CD8(+)αβ T-cell compartments compared with the equivalent tetramers. The low-affinity, tetramer-negative, dodecamer-positive T cells showed comparable effector cytokine responses as those of high-affinity, tetramer-positive T cells. Dodecamers are able to detect early stage CD4(+)CD8(+)double-positive thymocytes on which T-cell receptors are 10- to 30-fold less dense than mature T cells. Dodecamers also show utility in the analysis of γδ T cells and in cytometry by time-of-flight applications. This construct has a simple structure with a central scaffold protein linked to four streptavidin molecules, each having three pMHC ligands or other molecules. The dodecamer is straightforward and inexpensive to produce and is compatible with current tetramer technology and commercially available streptavidin conjugates.

  View details for DOI 10.1073/pnas.1602488113

  View details for Web of Science ID 000372876400017

  View details for PubMedCentralID PMC4822615

• Thermoelectric Power in Bilayer Graphene Device with Ionic Liquid Gating SCIENTIFIC REPORTS Chien, Y., Yuan, H., Wang, C., Lee, W. 2016; 6: 20402

  Abstract

  The quest for materials showing large thermoelectric power has long been one of the important subjects in material science and technology. Such materials have great potential for thermoelectric cooling and also high figure of merit ZT thermoelectric applications. We have fabricated bilayer graphene devices with ionic-liquid gating in order to tune its band gap via application of a perpendicular electric field on a bilayer graphene. By keeping the Fermi level at charge neutral point during the cool-down, we found that the charge puddles effect can be greatly reduced and thus largely improve the transport properties at low T in graphene-based devices using ionic liquid gating. At (Vig, Vbg) = (-1 V, +23 V), a band gap of about 36.6 ± 3 meV forms, and a nearly 40% enhancement of thermoelectric power at T = 120 K is clearly observed. Our works demonstrate the feasibility of band gap tuning in a bilayer graphene using ionic liquid gating. We also remark on the significant influence of the charge puddles effect in ionic-liquid-based devices.

  View details for PubMedID 26852799

• A highly focused antigen receptor repertoire characterizes gamma delta T cells that are poised to make IL-17 rapidly in naive animals FRONTIERS IN IMMUNOLOGY Wei, Y., Han, A., Glanville, J., Fang, F., Zunige, L. A., Lee, J. S., Cue, D. J., Chien, Y. 2015; 6: 1-6

  Abstract

  Interleukin (IL)-17 plays a key role in immunity. In acute infections, a rapid IL-17 response must be induced without prior antigen exposure, and γδ T cells are the major initial IL-17 producers. In fact, some γδ T cells make IL-17 within hours after an immune challenge. These cells appear to acquire the ability to respond to IL-1 and IL-23 and to make IL-17 naturally in naïve animals. They are known as the natural Tγδ17 (nTγδ17) cells. The rapidity of the nTγδ17 response, and the apparent lack of explicit T cell receptor (TCR) engagement for its induction have led to the view that this is a cytokine (IL-1, IL-23)-mediated response. However, pharmacological inhibition or genetic defects in TCR signaling drastically reduce the nTγδ17 response and/or their presence. To better understand antigen recognition in this rapid IL-17 response, we analyzed the antigen receptor repertoire of IL-1R(+)/IL-23R(+) γδ T cells, a proxy for nTγδ17 cells in naïve animals directly ex vivo, using a barcode-enabled high throughput single-cell TCR sequence analysis. We found that regardless of their anatomical origin, these cells have a highly focused TCR repertoire. In particular, the TCR sequences have limited V gene combinations, little or no junctional diversity and much reduced or no N region diversity. In contrast, IL-23R(-) cells at mucosal sites similar to most of the splenic γδ T cells and small intestine epithelial γδ lymphocytes expressed diverse TCRs. This remarkable commonality and restricted repertoire of IL-1R(+)/IL-23R(+) γδ T cells underscores the importance of antigen recognition in their establishment/function.

  View details for DOI 10.3389/fimmu.2015.00118

  View details for Web of Science ID 000354816300001

  View details for PubMedCentralID PMC4370043

• Pictionary-based fMRI paradigm to study the neural correlates of spontaneous improvisation and figural creativity. Scientific reports Saggar, M., Quintin, E., Kienitz, E., Bott, N. T., Sun, Z., Hong, W., Chien, Y., Liu, N., Dougherty, R. F., Royalty, A., Hawthorne, G., Reiss, A. L. 2015; 5: 10894-?

  Abstract

  A novel game-like and creativity-conducive fMRI paradigm is developed to assess the neural correlates of spontaneous improvisation and figural creativity in healthy adults. Participants were engaged in the word-guessing game of Pictionary(TM), using an MR-safe drawing tablet and no explicit instructions to be "creative". Using the primary contrast of drawing a given word versus drawing a control word (zigzag), we observed increased engagement of cerebellum, thalamus, left parietal cortex, right superior frontal, left prefrontal and paracingulate/cingulate regions, such that activation in the cingulate and left prefrontal cortices negatively influenced task performance. Further, using parametric fMRI analysis, increasing subjective difficulty ratings for drawing the word engaged higher activations in the left pre-frontal cortices, whereas higher expert-rated creative content in the drawings was associated with increased engagement of bilateral cerebellum. Altogether, our data suggest that cerebral-cerebellar interaction underlying implicit processing of mental representations has a facilitative effect on spontaneous improvisation and figural creativity.

  View details for DOI 10.1038/srep10894

  View details for PubMedID 26018874

• The Split Virus Influenza Vaccine rapidly activates immune cells through Fc gamma receptors VACCINE O'Gorman, W. E., Huang, H., Wei, Y., Davis, K. L., Leipold, M. D., Bendall, S. C., Kidd, B. A., Dekker, C. L., Maecker, H. T., Chien, Y., Davis, M. M. 2014; 32 (45): 5989-5997

  Abstract

  Seasonal influenza vaccination is one of the most common medical procedures and yet the extent to which it activates the immune system beyond inducing antibody production is not well understood. In the United States, the most prevalent formulations of the vaccine consist of degraded or "split" viral particles distributed without any adjuvants. Based on previous reports we sought to determine whether the split influenza vaccine activates innate immune receptors-specifically Toll-like receptors. High-dimensional proteomic profiling of human whole-blood using Cytometry by Time-of-Flight (CyTOF) was used to compare signaling pathway activation and cytokine production between the split influenza vaccine and a prototypical TLR response ex vivo. This analysis revealed that the split vaccine rapidly and potently activates multiple immune cell types but yields a proteomic signature quite distinct from TLR activation. Importantly, vaccine induced activity was dependent upon the presence of human sera indicating that a serum factor was necessary for vaccine-dependent immune activation. We found this serum factor to be human antibodies specific for influenza proteins and therefore immediate immune activation by the split vaccine is immune-complex dependent. These studies demonstrate that influenza virus "splitting" inactivates any potential adjuvants endogenous to influenza, such as RNA, but in previously exposed individuals can elicit a potent immune response by facilitating the rapid formation of immune complexes.

  View details for DOI 10.1016/j.vaccine.2014.07.115

  View details for Web of Science ID 000343629900016

  View details for PubMedCentralID PMC4191649

• The Split Virus Influenza Vaccine rapidly activates immune cells through Fc? receptors. Vaccine O'Gorman, W. E., Huang, H., Wei, Y., Davis, K. L., Leipold, M. D., Bendall, S. C., Kidd, B. A., Dekker, C. L., Maecker, H. T., Chien, Y., Davis, M. M. 2014; 32 (45): 5989-5997

  Abstract

  Seasonal influenza vaccination is one of the most common medical procedures and yet the extent to which it activates the immune system beyond inducing antibody production is not well understood. In the United States, the most prevalent formulations of the vaccine consist of degraded or "split" viral particles distributed without any adjuvants. Based on previous reports we sought to determine whether the split influenza vaccine activates innate immune receptors-specifically Toll-like receptors. High-dimensional proteomic profiling of human whole-blood using Cytometry by Time-of-Flight (CyTOF) was used to compare signaling pathway activation and cytokine production between the split influenza vaccine and a prototypical TLR response ex vivo. This analysis revealed that the split vaccine rapidly and potently activates multiple immune cell types but yields a proteomic signature quite distinct from TLR activation. Importantly, vaccine induced activity was dependent upon the presence of human sera indicating that a serum factor was necessary for vaccine-dependent immune activation. We found this serum factor to be human antibodies specific for influenza proteins and therefore immediate immune activation by the split vaccine is immune-complex dependent. These studies demonstrate that influenza virus "splitting" inactivates any potential adjuvants endogenous to influenza, such as RNA, but in previously exposed individuals can elicit a potent immune response by facilitating the rapid formation of immune complexes.

  View details for DOI 10.1016/j.vaccine.2014.07.115

  View details for PubMedID 25203448

• Gamma delta T cells recognize haptens and mount a hapten-specific response ELIFE Zeng, X., Meyer, C., Huang, J., Newell, E. W., Kidd, B. A., Wei, Y., Chien, Y. 2014; 3

  View details for DOI 10.7554/eLife.03609

  View details for Web of Science ID 000342126700006

• γδ T cells: first line of defense and beyond Annu Rev Immunol Chien, Y., Meyer, C., Bonneville, M. 2014
• Gamma delta T cells recognize haptens and mount a hapten-specific response. eLife Zeng, X., Meyer, C., Huang, J., Newell, E. W., Kidd, B. A., Wei, Y., Chien, Y. 2014; 3

  Abstract

  The ability to recognize small organic molecules and chemical modifications of host molecules is an essential capability of the adaptive immune system, which until now was thought to be mediated mainly by B cell antigen receptors. Here we report that small molecules, such as cyanine 3 (Cy3), a synthetic fluorescent molecule, and 4-hydroxy-3-nitrophenylacetyl (NP), one of the most noted haptens, are γδ T cell antigens, recognized directly by specific γδ TCRs. Immunization with Cy3 conjugates induces a rapid Cy3-specific γδ T cell IL-17 response. These results expand the role of small molecules and chemical modifications in immunity and underscore the role of γδ T cells as unique adaptive immune cells that couple B cell-like antigen recognition capability with T cell effector function.

  View details for DOI 10.7554/eLife.03609

  View details for PubMedID 25255099

  View details for PubMedCentralID PMC4174581

• gamma delta T Cells: First Line of Defense and Beyond ANNUAL REVIEW OF IMMUNOLOGY, VOL 32 Chien, Y., Meyer, C., Bonneville, M. 2014; 32: 121-155

  Abstract

  γδ T cells, αβ T cells, and B cells are present together in all but the most primitive vertebrates, suggesting that each population contributes to host immune competence uniquely and that all three are necessary for maintaining immune competence. Functional and molecular analyses indicate that in infections, γδ T cells respond earlier than αβ T cells do and that they emerge late after pathogen numbers start to decline. Thus, these cells may be involved in both establishing and regulating the inflammatory response. Moreover, γδ T cells and αβ T cells are clearly distinct in their antigen recognition and activation requirements as well as in the development of their antigen-specific repertoire and effector function. These aspects allow γδ T cells to occupy unique temporal and functional niches in host immune defense. We review these and other advances in γδ T cell biology in the context of their being the major initial IL-17 producers in acute infection.

  View details for DOI 10.1146/annurev-immunol-032713-120216

  View details for Web of Science ID 000336427400005

  View details for PubMedID 24387714

• A Single Peptide-Major Histocompatibility Complex Ligand Triggers Digital Cytokine Secretion in CD4(+) T Cells. Immunity Huang, J., Brameshuber, M., Zeng, X., Xie, J., Li, Q., Chien, Y., Valitutti, S., Davis, M. M. 2013; 39 (5): 846-857

  Abstract

  We have developed a single-molecule imaging technique that uses quantum-dot-labeled peptide-major histocompatibility complex (pMHC) ligands to study CD4(+) T cell functional sensitivity. We found that naive T cells, T cell blasts, and memory T cells could all be triggered by a single pMHC to secrete tumor necrosis factor-α (TNF-α) and interleukin-2 (IL-2) cytokines with a rate of ∼1,000, ∼10,000, and ∼10,000 molecules/min, respectively, and that additional pMHCs did not augment secretion, indicating a digital response pattern. We also found that a single pMHC localized to the immunological synapse induced the slow formation of a long-lasting T cell receptor (TCR) cluster, consistent with a serial engagement mechanism. These data show that scaling up CD4(+) T cell cytokine responses involves increasingly efficient T cell recruitment rather than greater cytokine production per cell.

  View details for DOI 10.1016/j.immuni.2013.08.036

  View details for PubMedID 24120362

  View details for PubMedCentralID PMC3846396

• Dietary gluten triggers concomitant activation of CD4(+) and CD8(+) alpha beta T cells and gamma delta T cells in celiac disease PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA Han, A., Newell, E. W., Glanville, J., Fernandez-Becker, N., Khosla, C., Chien, Y., Davis, M. M. 2013; 110 (32): 13073-13078

  View details for DOI 10.1073/pnas.1311861110

  View details for Web of Science ID 000322771100057

• Dietary gluten triggers concomitant activation of CD4+ and CD8+ aß T cells and ?d T cells in celiac disease. Proceedings of the National Academy of Sciences of the United States of America Han, A., Newell, E. W., Glanville, J., Fernandez-Becker, N., Khosla, C., Chien, Y., Davis, M. M. 2013; 110 (32): 13073-13078

  Abstract

  Celiac disease is an intestinal autoimmune disease driven by dietary gluten and gluten-specific CD4(+) T-cell responses. In celiac patients on a gluten-free diet, exposure to gluten induces the appearance of gluten-specific CD4(+) T cells with gut-homing potential in the peripheral blood. Here we show that gluten exposure also induces the appearance of activated, gut-homing CD8(+) αβ and γδ T cells in the peripheral blood. Single-cell T-cell receptor sequence analysis indicates that both of these cell populations have highly focused T-cell receptor repertoires, indicating that their induction is antigen-driven. These results reveal a previously unappreciated role of antigen in the induction of CD8(+) αβ and γδ T cells in celiac disease and demonstrate a coordinated response by all three of the major types of T cells. More broadly, these responses may parallel adaptive immune responses to viral pathogens and other systemic autoimmune diseases.

  View details for DOI 10.1073/pnas.1311861110

  View details for PubMedID 23878218

  View details for PubMedCentralID PMC3740842

• Correlation of Gene Expression and Genome Mutation in Single B-Cells PLOS ONE Weinstein, J. A., Zeng, X., Chien, Y., Quake, S. R. 2013; 8 (6)

  Abstract

  High-throughput measurement of gene-expression and immune receptor repertoires have recently become powerful tools in the study of adaptive immune response. However, despite their now-widespread use, both tend to discard cell identity by treating cell populations in bulk, and therefore lose the correlation between genetic variability and gene-expression at the single cell level. In order to recover this information, we developed a method to simultaneously measure gene expression profiles and genome mutations in single cells. We applied this method by quantifying the relationships between gene expression and antibody mutation in ensembles of individual B-cells from immunized mice. The results reveal correlations reflecting the manner in which information propagates between a B-cell's antigen receptors, its gene expression, and its mutagenic machinery, and demonstrate the power of this approach to illuminate both heterogeneity and physiology in cell populations.

  View details for DOI 10.1371/journal.pone.0067624

  View details for Web of Science ID 000321148400117

  View details for PubMedID 23840752

  View details for PubMedCentralID PMC3695916

• The natural and the inducible: interleukin (IL)-17-producing ?d T cells. Trends in immunology Chien, Y., Zeng, X., Prinz, I. 2013; 34 (4): 151-154

  Abstract

  γδ T cells are the major initial interleukin (IL)-17 producers in acute infections. Recent studies have indicated that some γδ T cells have IL-17-producing capabilities without explicit induction of an immune response. They are preferentially localized in barrier tissues and are likely to originate from fetal γδ thymocytes. In addition, γδ T cells present in the secondary lymphoid organs will mature and differentiate to produce IL-17 after antigen encounter in an immune response. Based on these studies, we propose that there are two different sets of IL-17-producing γδ T cells (Tγδ17) referred to as the 'natural' and the 'inducible' Tγδ17 cells. This review focuses on recent publications leading to the delineation of these two types of cells and their implied roles in host immune defense.

  View details for DOI 10.1016/j.it.2012.11.004

  View details for PubMedID 23266231

  View details for PubMedCentralID PMC3622789

• Correlation of gene expression and genome mutation in single B-cells. PloS one Weinstein, J. A., Zeng, X., Chien, Y., Quake, S. R. 2013; 8 (6)

  Abstract

  High-throughput measurement of gene-expression and immune receptor repertoires have recently become powerful tools in the study of adaptive immune response. However, despite their now-widespread use, both tend to discard cell identity by treating cell populations in bulk, and therefore lose the correlation between genetic variability and gene-expression at the single cell level. In order to recover this information, we developed a method to simultaneously measure gene expression profiles and genome mutations in single cells. We applied this method by quantifying the relationships between gene expression and antibody mutation in ensembles of individual B-cells from immunized mice. The results reveal correlations reflecting the manner in which information propagates between a B-cell's antigen receptors, its gene expression, and its mutagenic machinery, and demonstrate the power of this approach to illuminate both heterogeneity and physiology in cell populations.

  View details for DOI 10.1371/journal.pone.0067624

  View details for PubMedID 23840752

  View details for PubMedCentralID PMC3695916

• gamma delta T Cells Recognize a Microbial Encoded B Cell Antigen to Initiate a Rapid Antigen-Specific Interleukin-17 Response IMMUNITY Zeng, X., Wei, Y., Huang, J., Newell, E. W., Yu, H., Kidd, B. A., Kuhns, M. S., Waters, R. W., Davis, M. M., Weaver, C. T., Chien, Y. 2012; 37 (3): 524-534

  Abstract

  γδ T cells contribute uniquely to immune competence. Nevertheless, how they function remains an enigma. It is unclear what most γδ T cells recognize, what is required for them to mount an immune response, and how the γδ T cell response is integrated into host immune defense. Here, we report that a noted B cell antigen, the algae protein phycoerythrin (PE), is a murine and human γδ T cell antigen. Employing this specificity, we demonstrated that antigen recognition activated naive γδ T cells to make interleukin-17 and respond to cytokine signals that perpetuate the response. High frequencies of antigen-specific γδ T cells in naive animals and their ability to mount effector response without extensive clonal expansion allow γδ T cells to initiate a swift, substantial response. These results underscore the adaptability of lymphocyte antigen receptors and suggest an antigen-driven rapid response in protective immunity prior to the maturation of classical adaptive immunity.

  View details for DOI 10.1016/j.immuni.2012.06.011

  View details for Web of Science ID 000309199000016

  View details for PubMedID 22960222

  View details for PubMedCentralID PMC3495981

• Toxoplasma Polymorphic Effectors Determine Macrophage Polarization and Intestinal Inflammation CELL HOST & MICROBE Jensen, K. D., Wang, Y., Wojno, E. D., Shastri, A. J., Hu, K., Cornel, L., Boedec, E., Ong, Y., Chien, Y., Hunter, C. A., Boothroyd, J. C., Saeij, J. P. 2011; 9 (6): 472-483

  Abstract

  European and North American strains of the parasite Toxoplasma gondii belong to three distinct clonal lineages, type I, type II, and type III, which differ in virulence. Understanding the basis of Toxoplasma strain differences and how secreted effectors work to achieve chronic infection is a major goal of current research. Here we show that type I and III infected macrophages, a cell type required for host immunity to Toxoplasma, are alternatively activated, while type II infected macrophages are classically activated. The Toxoplasma rhoptry kinase ROP16, which activates STAT6, is responsible for alternative activation. The Toxoplasma dense granule protein GRA15, which activates NF-κB, promotes classical activation by type II parasites. These effectors antagonistically regulate many of the same genes, and mice infected with type II parasites expressing type I ROP16 are protected against Toxoplasma-induced ileitis. Thus, polymorphisms in determinants that modulate macrophage activation influence the ability of Toxoplasma to establish a chronic infection.

  View details for DOI 10.1016/j.chom.2011.04.015

  View details for Web of Science ID 000293157200006

  View details for PubMedID 21669396

  View details for PubMedCentralID PMC3131154

• Ligand recognition during thymic development and gamma delta T cell function specification SEMINARS IN IMMUNOLOGY Meyer, C., Zeng, X., Chien, Y. 2010; 22 (4): 207-213

  Abstract

  gammadelta T cells develop in the thymus before entering the periphery. Recent work suggests that thymic development does little to constrain gammadelta T cell antigen specificities, but instead determines their effector fate. When triggered through the T cell receptor, ligand-naïve gammadelta T cells produce IL-17, ligand-experienced cells make IFN-gamma and those that are strongly self-reactive make IL-4. Importantly, gammadelta T cells are able to make cytokines immediately upon TCR engagement. These characteristics allow gammadelta T cells to initiate an acute inflammatory response to pathogens and to host antigens revealed by injury. These advances warrant a fresh look at how gammadelta T cells may function in the immune system.

  View details for DOI 10.1016/j.smim.2010.04.002

  View details for Web of Science ID 000280383400004

  View details for PubMedID 20430644

  View details for PubMedCentralID PMC2906674

• Evidence for a functional sidedness to the alpha beta TCR PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA Kuhns, M. S., Girvin, A. T., Klein, L. O., Chen, R., Jensen, K. D., Newell, E. W., Huppa, J. B., Lillemeier, B. F., Huse, M., Chien, Y., Garcia, K. C., Davis, M. M. 2010; 107 (11): 5094-5099

  Abstract

  The T cell receptor (TCR) and associated CD3gammaepsilon, deltaepsilon, and zetazeta signaling dimers allow T cells to discriminate between different antigens and respond accordingly, but our knowledge of how these parts fit and work together is incomplete. In this study, we provide additional evidence that the CD3 heterodimers congregate on one side of the TCR in both the alphabeta and gammadeltaTCR-CD3 complexes. We also report that the other side of the alphabetaTCR mediates homotypic alphabetaTCR interactions and signaling. Specifically, an erythropoietin receptor-based dimerization assay was used to show that, upon complex assembly, the CD3epsilon chains of two CD3 heterodimers are arranged side-by-side in both the alphabeta and gammadeltaTCR-CD3 complexes. This system was also used to show that alphabetaTCRs can dimerize in the cell membrane and that mutating the unusual outer strands of the Calpha domain impairs this dimerization. Finally, we present data showing that, for CD4 T cells, the mutations that impair alphabetaTCR dimerization also alter ligand-induced calcium mobilization, TCR accumulation at the site of pMHC contact, and polarization toward the site of antigen contact. These data reveal a "functional-sidedness" to the alphabetaTCR constant region, with dimerization occurring on the side of the TCR opposite from where the CD3 heterodimers are located.

  View details for DOI 10.1073/pnas.1000925107

  View details for Web of Science ID 000275714300054

  View details for PubMedID 20202921

  View details for PubMedCentralID PMC2841884

• The Salmonella SPI2 Effector SseI Mediates Long-Term Systemic Infection by Modulating Host Cell Migration PLOS PATHOGENS McLaughlin, L. M., Govoni, G. R., Gerke, C., Gopinath, S., Peng, K., Laidlaw, G., Chien, Y., Jeong, H., Li, Z., Brown, M. D., Sacks, D. B., Monack, D. 2009; 5 (11)

  Abstract

  Host-adapted strains of Salmonella enterica cause systemic infections and have the ability to persist systemically for long periods of time despite the presence of a robust immune response. Chronically infected hosts are asymptomatic and transmit disease to naïve hosts via fecal shedding of bacteria, thereby serving as a critical reservoir for disease. We show that the bacterial effector protein SseI (also called SrfH), which is translocated into host cells by the Salmonella Pathogenicity Island 2 (SPI2) type III secretion system (T3SS), is required for Salmonella typhimurium to maintain a long-term chronic systemic infection in mice. SseI inhibits normal cell migration of primary macrophages and dendritic cells (DC) in vitro, and such inhibition requires the host factor IQ motif containing GTPase activating protein 1 (IQGAP1), an important regulator of cell migration. SseI binds directly to IQGAP1 and co-localizes with this factor at the cell periphery. The C-terminal domain of SseI is similar to PMT/ToxA, a bacterial toxin that contains a cysteine residue (C1165) that is critical for activity. Mutation of the corresponding residue in SseI (C178A) eliminates SseI function in vitro and in vivo, but not binding to IQGAP1. In addition, infection with wild-type (WT) S. typhimurium suppressed DC migration to the spleen in vivo in an SseI-dependent manner. Correspondingly, examination of spleens from mice infected with WT S. typhimurium revealed fewer DC and CD4(+) T lymphocytes compared to mice infected with Delta sseI S. typhimurium. Taken together, our results demonstrate that SseI inhibits normal host cell migration, which ultimately counteracts the ability of the host to clear systemic bacteria.

  View details for DOI 10.1371/journal.ppat.1000671

  View details for Web of Science ID 000274226600025

  View details for PubMedID 19956712

  View details for PubMedCentralID PMC2777311

• Cutting Edge: gamma delta Intraepithelial Lymphocytes of the Small Intestine Are Not Biased toward Thymic Antigens JOURNAL OF IMMUNOLOGY Jensen, K. D., Shin, S., Chien, Y. 2009; 182 (12): 7348-7351

  Abstract

  gammadelta Tau cells, together with alphabeta Tau cells, are abundantly present in the epithelial layer of the small intestine (IEL) and are essential for the host's first line of defense. Whether or not gammadelta IELs, like alphabeta IELs, are derived from thymocytes that encounter self-Ags in the thymus is unclear. In this study, we report that a natural population of gammadelta T cells that are specific for the nonclassical MHC class I molecules T10 and T22 are present in the IEL compartment of mice that do not express T10/T22. Furthermore, the small intestinal homing receptor CCR9 is preferentially expressed on gammadelta thymocytes that have yet to encounter a ligand, and gammadelta thymocytes with high affinity for self-ligand are CCR9(low). These observations suggest that the Ag-specific repertoire of gammadelta IELs is not biased toward thymic Ags. Instead, gammadelta IELs appear suited to respond to novel Ags revealed in pathological settings.

  View details for DOI 10.4049/jimmunol.0900465

  View details for Web of Science ID 000266833900003

  View details for PubMedID 19494256

• Thymic maturation determines gamma delta T cell function, but not their antigen specificities CURRENT OPINION IN IMMUNOLOGY Jensen, K. D., Chien, Y. 2009; 21 (2): 140-145

  Abstract

  gammadelta T cells contribute uniquely to host immune defense, but how they do so remains unclear. Recent work suggests that thymic selection does little to constrain gammadelta T cell antigen specificities, but instead determines their effector fate. When triggered through the T cell receptor, ligand-experienced cells make IFNgamma, whereas ligand-naïve gammadelta T cells produce IL-17, a major initiator of inflammation. These advances warrant a fresh look at how gammadelta T cells may function in the immune system.

  View details for DOI 10.1016/j.coi.2009.02.008

  View details for Web of Science ID 000266223400005

  View details for PubMedID 19321327

  View details for PubMedCentralID PMC2697822

• Thymic selection determines gamma delta T cell effector fate: Antigen-naive cells make interleukin-17 and antigen-experienced cells make interferon gamma IMMUNITY Jensen, K. D., Su, X., Shin, S., Li, L., Youssef, S., Yarnasaki, S., Steinman, L., Saito, T., Locksley, R. M., Davis, M. M., Baumgarth, N., Chien, Y. 2008; 29 (1): 90-100

  Abstract

  gammadelta T cells uniquely contribute to host immune defense, but how this is accomplished remains unclear. Here, we analyzed the nonclassical major histocompatibility complex class I T10 and T22-specific gammadelta T cells in mice and found that encountering antigen in the thymus was neither required nor inhibitory for their development. But when triggered through the T cell receptor, ligand-naive lymphoid-gammadelta T cells produced IL-17, whereas ligand-experienced cells made IFN-gamma. Immediately after immunization, a large fraction of IL-17(+) gammadelta T cells were found in the draining lymph nodes days before the appearance of antigen-specific IL-17(+) *beta T cells. Thus, thymic selection determines the effector fate of gammadelta T cells rather than constrains their antigen specificities. The swift IL-17 response mounted by antigen-naive gammadelta T cells suggests a critical role for these cells at the onset of an acute inflammatory response to novel antigens.

  View details for DOI 10.1016/j.immuni.2008.04.022

  View details for Web of Science ID 000257905400013

  View details for PubMedID 18585064

  View details for PubMedCentralID PMC2601709

• An autonomous CDR3 delta is sufficient for recognition of the nonclassical MHC class I molecules T10 and T22 by gamma delta T cells NATURE IMMUNOLOGY Adams, E. J., Strop, P., Shin, S., Chien, Y., Garcia, K. C. 2008; 9 (7): 777-784

  Abstract

  It remains unclear whether gammadelta T cell antigen receptors (TCRs) detect antigens in a way similar to antibodies or alphabeta TCRs. Here we show that reactivity between the G8 and KN6 gammadelta TCRs and the major histocompatibility complex class Ib molecule T22 could be recapitulated, with retention of wild-type ligand affinity, in an alphabeta TCR after grafting of a G8 or KN6 complementarity-determining region 3-delta (CDR3delta) loop in place of the CDR3alpha loop of an alphabeta TCR. We also found that a shared sequence motif in CDR3delta loops of all T22-reactive gammadelta TCRs bound T22 in energetically distinct ways, and that T10(d), which bound G8 with weak affinity, was converted into a high-affinity ligand by a single point mutation. Our results demonstrate unprecedented autonomy of a single CDR3 loop in antigen recognition.

  View details for DOI 10.1038/ni.1620

  View details for Web of Science ID 000256904900016

  View details for PubMedID 18516039

  View details for PubMedCentralID PMC2768525

• Antigen recognition by gamma delta T cells IMMUNOLOGICAL REVIEWS Chien, Y., Konigshofer, Y. 2007; 215: 46-58

  Abstract

  gammadelta T cells contribute to host immune competence uniquely. This is most likely because they have distinctive antigen-recognition properties. While the basic organization of gammadelta T-cell receptor (TCR) loci is similar to that of alphabeta TCR loci, there is a striking difference in how the diversity of gammadelta TCRs is generated. gammadelta and alphabeta T cells have different antigen-recognition requirements and almost certainly recognize a different set of antigens. While it is unclear what most gammadelta T cells recognize, the non-classical major histocompatibility complex class I molecules T10 and T22 were found to be the natural ligands for a sizable population (0.2-2%) of murine gammadelta T cells. The recognition of T10/T22 may be a way by which gammadelta T cells regulate cells of the immune system, and this system has been used to determine the antigen-recognition determinants of gammadelta T cells. T10/T22-specific gammadelta T cells have TCRs that are diverse in both V gene usage and CDR3 sequences. Their Vgamma usage reflects their tissue origin, and their antigen specificity is conferred by a motif in the TCR delta chain that is encoded by V and D segments and by P-nucleotide addition. Sequence variations around this motif modulate affinities between TCRs and T10/T22. That this CDR3 motif is important in antigen recognition is confirmed by the crystal structure of a gammadelta TCR bound to its ligand. The significance of these observations is discussed in the context of gammadelta T-cell biology.

  View details for Web of Science ID 000243973600005

  View details for PubMedID 17291278

• gamma delta T cells - innate immune lymphocytes? CURRENT OPINION IN IMMUNOLOGY Konigshofer, Y., Chien, Y. 2006; 18 (5): 527-533

  Abstract

  It is unclear what the antigen recognition determinants of gammadelta T-cell receptors (TCRs) are. Compared with immunoglobulin and alphabeta TCRs, gammadelta TCRs have the highest potential CDR3 diversity generated by VDJ recombination. However, gammadelta T-cell reactivities seem to segregate with V gene usage, which has been taken to suggest that rearrangement has little role in generating different antigen specificities. During the past year, the CDR3 regions were found to determine the antigen specificities of T10- and T22-reactive gammadelta TCRs, a surface protein complex was identified as a ligand for human phosphoantigen-reactive gammadelta T cells, and the first co-crystal structure of a gammadelta TCR bound to its ligand was reported. These advances warrant a fresh look at gammadelta T-cell antigen recognition.

  View details for DOI 10.1016/j.coi.2006.07.008

  View details for Web of Science ID 000240816300003

  View details for PubMedID 16879956

• Nonobese diabetic mice express aspects of both type 1 and type 2 diabetes PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA Chaparro, R. J., Konigshofer, Y., Beilhack, G. F., Shizuru, J. A., McDevitt, H. O., Chien, Y. 2006; 103 (33): 12475-12480

  Abstract

  Before the onset of autoimmune destruction, type 1 diabetic patients and an animal model, the nonobese diabetic (NOD) mouse, show morphological and functional abnormalities in target organs, which may act as inciting events for leukocyte infiltration. To better understand these abnormalities, but without the complications associated with lymphocytic infiltrates, we examined genes expressed in autoimmune target tissues of NOD/severe combined immunodeficient (scid) mice and of autoimmune-resistant C57BL/6/scid mice. Our results suggest that the NOD genetic background may predispose them to diabetic complications, including insulin resistance in the absence of high circulating glucose levels and without autoimmune destruction of their beta cells. Several of these genes lie within known type 1 and 2 diabetes loci. These data suggest that the NOD mouse may be a good candidate to study an interface between type 1 and type 2 diabetes.

  View details for DOI 10.1073/pnas.0604317103

  View details for Web of Science ID 000239867500050

  View details for PubMedID 16895987

  View details for PubMedCentralID PMC1832259

• The shaping of the gamma delta T cell receptor repertoire Annual Meeting of the American-Association-of-Immunologists Jensen, K., Shin, S., Konigshofer, Y., Li, L., Chien, Y. AMER ASSOC IMMUNOLOGISTS. 2006: S320–S320

  View details for Web of Science ID 000238837102491

• Structure of a gamma delta T cell receptor in complex with the nonclassical MHC T22 SCIENCE Adams, E. J., Chien, Y. H., Garcia, K. C. 2005; 308 (5719): 227-231

  Abstract

  Gammadelta T cell receptors (TCRs), alphabeta TCRs, and antibodies are the three lineages of somatically recombined antigen receptors. The structural basis for ligand recognition is well defined for alphabeta TCR and antibodies but is lacking for gammadelta TCRs. We present the 3.4 A structure of the murine gammadelta TCR G8 bound to its major histocompatibility complex (MHC) class Ib ligand, T22. G8 predominantly uses germline-encoded residues of its delta chain complementarity-determining region 3 (CDR3) loop to bind T22 in an orientation substantially different from that seen in alphabeta TCR/peptide-MHC. That junctionally encoded G8 residues play an ancillary role in binding suggests a fusion of innate and adaptive recognition strategies.

  View details for DOI 10.1126/science.1106885

  View details for Web of Science ID 000228273700045

  View details for PubMedID 15821084

• Antigen recognition determinants of gamma delta T cell receptors SCIENCE Shin, S., El-Diwany, R., Schaffert, S., Adams, E. J., Garcia, K. C., Pereira, P., Chien, Y. H. 2005; 308 (5719): 252-255

  Abstract

  The molecular basis of gammadelta T cell receptor (TCR) recognition is poorly understood. Here, we analyze the TCR sequences of a natural gammadelta T cell population specific for the major histocompatibility complex class Ib molecule T22. We find that T22 recognition correlates strongly with a somatically recombined TCRdelta complementarity-determining region 3 (CDR3) motif derived from germ line-encoded residues. Sequence diversity around these residues modulates TCR ligand-binding affinities, whereas V gene usage correlates mainly with tissue origin. These results show how an antigen-specific gammadelta TCR repertoire can be generated at a high frequency and suggest that gammadelta T cells recognize a limited number of antigens.

  View details for DOI 10.1126/science.1106480

  View details for Web of Science ID 000228273700053

  View details for PubMedID 15821090

• The adaptor molecules LAT and SLP-76 are specifically targeted by Yersinia to inhibit T cell activation JOURNAL OF EXPERIMENTAL MEDICINE Gerke, C., FALKOW, S., Chien, Y. H. 2005; 201 (3): 361-371

  Abstract

  T cell responses are critical to the survival of Yersinia-infected animals. Yersinia have the ability to directly suppress T lymphocyte activation through the virulence factor YopH, a tyrosine phosphatase. Using single cell video microscopy and FACS analysis, here we show that even an average of one Yersinia per T cell is sufficient to inhibit or alter T cell responses. This efficient inhibition is traced to specific targeting by YopH of the adaptor proteins, linker for activation of T cells (LAT) and SH2-domain-containing leukocyte protein of 76 kD (SLP-76), which are crucial for T cell antigen receptor (TCR) signaling. A catalytically inactive YopH translocated via the type III secretory pathway from the bacteria into T cells primarily binds to LAT and SLP-76. Furthermore, among the proteins of the TCR signaling pathway, the tyrosine phosphorylation levels of LAT and SLP-76 are the most affected in T cells exposed to low numbers of Yersinia pseudotuberculosis. This is the first example showing that a pathogen targets these adaptor proteins in the TCR signaling pathway, suggesting a novel mechanism by which pathogens may efficiently alter T cell-mediated immune responses.

  View details for DOI 10.1084/jem.20041120

  View details for Web of Science ID 000227092900007

  View details for PubMedID 15699071

  View details for PubMedCentralID PMC2213036

• Autoimmune destruction in the CNS triggered by non-classical MHC class I-specific gamma delta T cells 9th Annual Meeting of Americas-Committee-for-Treatment-and-Research-in-Multiple-Sclerosis Yeh, J. H., Youssef, S., Chan, J. R., Cosgaya, J. M., Steinman, L., Chien, Y. H. SAGE PUBLICATIONS LTD. 2005: 110–11

  View details for Web of Science ID 000226831700029

• Attributes of gamma delta intraepithelial lymphocytes as suggested by their transcriptional profile PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA Fahrer, A. M., Konigshofer, Y., Kerr, E. M., Ghandour, G., Mack, D. H., DAVIS, M. M., Chien, Y. H. 2001; 98 (18): 10261-10266

  Abstract

  gammadelta T lymphocytes in the intestinal intraepithelial layer (gammadelta IELs) are thought to contribute to immune competence, but their actual function remains poorly understood. Here we used DNA microarrays to study the gene expression profile of gammadelta IELs in a Yersinia infection system to better define their roles. To validate this approach, mesenteric lymph node CD8(+) alphabeta T cells were similarly analyzed. The transcription profiles show that, whereas lymph node CD8(+) alphabeta T cells must be activated to become cytotoxic effectors, gammadelta IELs are constitutively activated and appear to use different signaling cascades. Our data suggest that gammadelta IELs may respond efficiently to a broad range of pathological situations irrespective of their diverse T cell antigen receptor repertoire. gammadelta IELs may modulate local immune responses and participate in intestinal lipid metabolism, cholesterol homeostasis, and physiology. This study provides a strong basis for further investigations of the roles of these cells as well as mucosal immune defense in general.

  View details for Web of Science ID 000170738000045

  View details for PubMedID 11526237

• Differential roles of cytokine receptors in the development of epidermal gamma delta T cells JOURNAL OF IMMUNOLOGY Ye, S. K., Maki, K., Lee, H. C., Ito, A., Kawai, K., Suzuki, H., Mak, T. W., Chien, Y. H., Honjo, T., Ikuta, K. 2001; 167 (4): 1929-1934

  Abstract

  IL-7 and IL-15 play important roles in gammadelta T cell development. These receptors transmit proliferation and/or survival signals in gammadelta T cells. In addition, the IL-7R promotes recombination and transcription in the TCR gamma locus. To clarify the role of the cytokine receptors in the development of epidermal gammadelta T cells, we introduced a Vgamma3/Vdelta1 TCR transgene, derived from Thy-1+ dendritic epidermal T cells (DETC), into IL-7Ralpha-deficient mice, and we found that they partly rescued gammadelta T cells in the adult thymus but not in the spleen. Introduction of an additional Bcl-2 transgene had a minimal effect on gammadelta T cells in the adult thymus of these mice. In contrast to the adult thymus, the introduction of the Vgamma3/Vdelta1 TCR transgene into IL-7Ralpha-/- mice completely restored Vgamma3+ T cells in the fetal thymus and DETC in the adult skin. On the contrary, the same Vgamma3/Vdelta1 TCR transgene failed to rescue DETC in the skin of IL-2Rbeta-deficient mice, even with the additional Bcl-2 transgene. These results suggest that the IL-2/IL-15R, rather than the IL-7R, plays an essential role in proliferation and survival of DETC in the fetal thymus and the skin. In contrast, the IL-7R is probably essential in the induction of V-J recombination of TCRgamma genes. Thus, this study proves that IL-7R and IL-2/IL-15R serve differential functions in epidermal gammadelta T cell development.

  View details for Web of Science ID 000170949600014

  View details for PubMedID 11489972

• Antigen-recognition properties of murine gamma delta T cells SPRINGER SEMINARS IN IMMUNOPATHOLOGY Chien, Y. H., Hampl, J. 2000; 22 (3): 239-250

  View details for Web of Science ID 000165436700005

  View details for PubMedID 11116955

• A population of murine gamma delta T cells that recognize an inducible MHC class lb molecule SCIENCE Crowley, M. P., Fahrer, A. M., Baumgarth, N., Hampl, J., Gutgemann, I., Teyton, L., Chien, Y. H. 2000; 287 (5451): 314-316

  Abstract

  Although gammadelta T cells are implicated in regulating immune responses, gammadelta T cell-ligand pairs that could mediate such regulatory functions have not been identified. Here, the expression of the major histocompatibility complex (MHC) class Ib T22 and the closely related T10 molecules is shown to be activation-induced, and they confer specificity to about 0.4% of the gammadelta T cells in normal mice. Thus, the increased expression of T22 and/or T10 might trigger immunoregulatory gammadelta T cells during immune responses. Furthermore, the fast on-rates and slow off-rates that characterize this receptor/ligand interaction would compensate for the low ligand stability and suggest a high threshold for gammadelta T cell activation.

  View details for Web of Science ID 000084769600043

  View details for PubMedID 10634788

• Crystal structure of a gamma delta T cell receptor ligand T22: A truncated MHC-like fold SCIENCE Wingren, C., Crowley, M. P., Degano, M., Chien, Y. H., Wilson, I. A. 2000; 287 (5451): 310-314

  Abstract

  Murine T10 and T22 are highly related nonclassical major histocompatibility complex (MHC) class Ib proteins that bind to certain gammadelta T cell receptors (TCRs) in the absence of other components. The crystal structure of T22b at 3.1 angstroms reveals similarities to MHC class I molecules, but one side of the normal peptide-binding groove is severely truncated, which allows direct access to the beta-sheet floor. Potential gammadelta TCR-binding sites can be inferred from functional mapping of T10 and T22 point mutants and allelic variants. Thus, T22 represents an unusual variant of the MHC-like fold and indicates that gammadelta and alphabeta TCRs interact differently with their respective MHC ligands.

  View details for Web of Science ID 000084769600042

  View details for PubMedID 10634787

• Suppression of T and B lymphocyte activation by a Yersinia pseudotuberculosis virulence factor, YopH JOURNAL OF EXPERIMENTAL MEDICINE Yao, T., Mecsas, J., Healy, J. I., FALKOW, S., Chien, Y. H. 1999; 190 (9): 1343-1350

  Abstract

  The acquired immune responses are crucial to the survival of Yersinia-infected animals. Mice lacking T cells are sensitive to Yersinia infection, and a humoral response to Yersinia can be protective. Diverse mechanisms for Yersinia to impair and evade the host innate immune defense have been suggested, but the effects of Yersinia on lymphocytes are not known. Here, we demonstrate that after a transient exposure to Y. pseudotuberculosis, T and B cells are impaired in their ability to be activated through their antigen receptors. T cells are inhibited in their ability to produce cytokines, and B cells are unable to upregulate surface expression of the costimulatory molecule, B7.2, in response to antigenic stimulation. The block of lymphocyte activation results from the inhibition of early phosphorylation events of the antigen receptor signaling complex. Through the use of Y. pseudotuberculosis mutants, we show that the inhibitory effect in both T cells and B cells is dependent on the production of Yersinia outermembrane protein (Yop) H, a tyrosine phosphatase. Our results suggest a mechanism by which the pathogenic bacteria may modulate a wide range of T and B cell-mediated immune responses.

  View details for Web of Science ID 000083552800014

  View details for PubMedID 10544205

• The specificity of a weak gamma delta TCR interaction can be modulated by the glycosylation of the ligand JOURNAL OF IMMUNOLOGY Hampl, J., SCHILD, H., Litzenberger, C., Baron, M., Crowley, M. P., Chien, Y. H. 1999; 163 (1): 288-294

  Abstract

  The gamma delta T cell clone LBK5 recognizes the MHC molecule IEk. Here, we demonstrate that the affinity of this interaction is weaker than those typically reported for alpha beta TCRs that recognize peptide/MHC complexes. Consistent with our previous finding that peptide bound to the IE molecule does not confer specificity, we show that the entire epitope for LBK5 is contained within the polypeptide chains of the molecule, centered around the polymorphic residues 67 and 70 of the IE beta-chain. However, LBK5 recognition is profoundly influenced by the N-linked glycosylation at residue 82 of the IE alpha-chain. Since infected, stressed, or transformed cells often change the posttranslational modifications of their surface glycoproteins, this finding suggests a new way in which gamma delta T cell Ag recognition can be regulated.

  View details for Web of Science ID 000080973700038

  View details for PubMedID 10384127

• Visualizing lymphocyte recognition IMMUNOLOGY AND CELL BIOLOGY Wulfing, C., Chien, Y. H., DAVIS, M. M. 1999; 77 (2): 186-187

  Abstract

  Studies of T cell recognition have entered new territory now that some of the basic issues of genetics, biochemistry and structure have been addressed, at least in outline form. In the present work, the focus is on a new aspect of T cell recognition that goes beyond classical biochemistry to ask, how to TCR and other cell surface molecules cooperate to initiate and control recognition?'

  View details for Web of Science ID 000079676100012

  View details for PubMedID 10234556

• Visualizing T-cell recognition 64th Symposia: Signaling and Gene Expression in the Immune System DAVIS, M. M., Wulfing, C., Krummel, M. F., Savage, P. A., Xu, J., Sumen, C., Dustin, M. L., Chien, Y. H. COLD SPRING HARBOR LAB PRESS, PUBLICATIONS DEPT. 1999: 243–251

  View details for Web of Science ID 000087225400030

  View details for PubMedID 11232292

• Differential effect of B lymphocyte-induced maturation protein (Blimp-1) expression on cell fate during B cell development JOURNAL OF EXPERIMENTAL MEDICINE Messika, E. J., Lu, P. S., Sung, Y. J., Yao, T., Chi, J. T., Chien, Y. H., DAVIS, M. M. 1998; 188 (3): 515-525

  Abstract

  The B lymphocyte-induced maturation protein (Blimp-1) upregulates the expression of syndecan-1 and J chain and represses that of c-myc. We have transfected Blimp-1 into two sublines of the BCL1 B cell lymphoma that represent distinct stages of B cell development in secondary lymphoid tissues. After interleukin (IL)-2 and IL-5 stimulation, the BCL1 3B3 cells differentiate into centrocyte-like cells, whereas the BCL1 5B1b cells blast and appear to be blocked at the centroblast stage. This blasting effect and the increase in IgM secretion that follows it can be blocked by a dominant negative form of Blimp-1. At the same time, the ectopic expression of Blimp-1 in these partially activated cells induces an apoptotic response that also can be suppressed by the same dominant negative protein. A similar effect was noticed when Blimp-1 was expressed in the mature L10A and the immature WEHI-231 lines, indicating this may be a general effect at earlier stages of the B cell development, and distinct from the ability of Blimp-1 to induce maturation in late stages of differentiation. Truncation mutants indicate that the induction of the apoptotic response relies mainly on 69 amino acids within Blimp-1's proline-rich domain. We propose that Blimp-1 expression defines a checkpoint beyond which fully activated B cells proceed to the plasma cell stage, whereas immature and partially activated cells are eliminated at this point.

  View details for Web of Science ID 000075300300010

  View details for PubMedID 9687529

• Induction of rapid T cell activation and tolerance by systemic presentation of an orally administered antigen IMMUNITY Gutgemann, I., Fahrer, A. M., Altman, J. D., DAVIS, M. M., Chien, Y. H. 1998; 8 (6): 667-673

  Abstract

  To understand how orally introduced antigen regulates peripheral immune responses, we fed cytochrome c protein to mice transgenic for the beta chain of a cytochrome c-specific TCR and followed the antigen-specific T cell responses with a cyt c/I-Ek tetramer staining reagent. We find that within 6 hr of cytochrome c administration, antigen-specific systemic T cell activation is induced, and spleen cells gain the ability to stimulate cytochrome c-specific T cell responses. Feeding multiple low doses of cytochrome c down-regulates the systemic immune response, which can be correlated with a reduction of antigen-specific T cells and not with immune deviation. These results suggest that systemic distribution of antigen contributes significantly to oral tolerance induction.

  View details for Web of Science ID 000074396300002

  View details for PubMedID 9655480

• Ligand recognition by alpha beta T cell receptors ANNUAL REVIEW OF IMMUNOLOGY Davis, M. M., BONIFACE, J. J., Reich, Z., Lyons, D., Hampl, J., Arden, B., Chien, Y. H. 1998; 16: 523-?

  Abstract

  While still incomplete, the first data concerning the biochemistry of T cell receptor-ligand interactions in cell-free systems seem to have considerable predictive value regarding whether a T cell response is strong or weak or suppressive. This data will help considerably in elucidating the mechanisms behind T cell responsiveness. Also of great interest are the first structures of T cell receptor molecules and, particularly, TCR-ligand complexes. These appear to confirm earlier suggestions of a fixed orientation for TCR engagement with peptide/MHC and should form the basis for understanding higher oligomers, evidence for which has also just emerged. We conclude with an analysis of the highly diverse CDR3 loops found in all antigen receptor molecules and suggest that such regions form the core of both TCR and antibody specificity.

  View details for Web of Science ID 000073129400019

  View details for PubMedID 9597140

• CD4 augments the response of a T cell to agonist but not to antagonist ligands IMMUNITY Hampl, J., Chien, Y. H., DAVIS, M. M. 1997; 7 (3): 379-385

  Abstract

  The recognition of peptide variants by the T cell receptor (TCR) has revealed a wide range of possible responses. Here, using a series of CD4+ and CD4- variants of the same T cell hybridoma, we find that while the expression of CD4 converts weak agonists into full agonists, none of the antagonist peptides are efficiently recognized as agonists. Furthermore, in antagonist assays, little difference can be seen in the response of CD4+ and CD4- T cells. Together with previous work showing a marked difference in stability between TCR binding to agonist versus antagonist ligands, these data suggest that CD4 engagement occurs after a TCR-peptide/MHC complex has formed and that it requires a certain minimal half-life of the ternary complex to be fully engaged in signaling.

  View details for Web of Science ID A1997XY83000007

  View details for PubMedID 9324358

• The recognition of the nonclassical major histocompatibility complex (MHC) class I molecule, T10, by the gamma delta T cell, G8 JOURNAL OF EXPERIMENTAL MEDICINE Crowley, M. P., Reich, Z., Mavaddat, N., Altman, J. D., Chien, Y. H. 1997; 185 (7): 1223-1230

  Abstract

  Recent studies have shown that many nonclassical major histocompatibility complex (MHC) (class 1b) molecules have distinct antigen-binding capabilities, including the binding of nonpeptide moieties and the binding of peptides that are different from those bound to classical MHC molecules. Here, we show that one of the H-2T region-encoded molecules, T10, when produced in Escherichia coli, can be folded in vitro with beta2-microglobulin (beta2m) to form a stable heterodimer in the absence of peptide or nonpeptide moieties. This heterodimer can be recognized by specific antibodies and is stimulatory to the gammadelta T cell clone, G8. Circular dichroism analysis indicates that T10/beta2m has structural features distinct from those of classical MHC class I molecules. These results suggest a new way for MHC-like molecules to adopt a peptide-free structure and to function in the immune system.

  View details for Web of Science ID A1997WU18300008

  View details for PubMedID 9104809

• T cell receptor biochemistry, repertoire selection and general features of TCR and Ig structure Symposium on the Molecular Basis of Cellular Defence Mechanisms DAVIS, M. M., Lyons, D. S., Altman, J. D., MCHEYZERWILLIAMS, M., Hampl, J., BONIFACE, J. J., Chien, Y., Nossal, Zinkernagel, Miller, Paul, Kirberg, MELCHERS, Mosmann, Goodnow JOHN WILEY & SONS LTD. 1997: 94–104

  Abstract

  T cell recognition is a central event in the development of most immune responses, whether appropriate or inappropriate (i.e. autoimmune). We are interested in reducing T cell recognition to its most elemental components and relating this to biological outcome. In a model system involving a cytochrome c-specific I-Ek restricted T cell receptor (TCR) derived from the 2B4 hybridoma, we have studied the interaction of soluble TCR and soluble peptide-MHC complexes using surface plasmon resonance. We find a striking continuum in which biological activity correlates best with the dissociation rate of the TCR from the peptide-MHC complex. In particular, we have found that weak agonists have significantly faster off-rates than strong agonists and that antagonists have even faster off-rates. This suggests that the stability of TCR binding to a given ligand is critically important with respect to whether the T cell is stimulated, inhibited or remains indifferent. It also suggests that the phenomenon of peptide antagonists might be explained purely by kinetic models and that conformation, either inter- or intramolecular, may not be a factor. We have also studied TCR repertoire selection during the establishment of a cytochrome c response, initially using an anti-TCR antibody strategy, but more recently using peptide-MHC tetramers as antigen-specific staining reagents. These tetramers work well with either class I or class II MHC-specific TCRs and have many possible applications. Lastly, we have also tried to correlate the structural and genetic features of TCRs with their function. Recent data on TCR structure as well as previous findings with antibodies suggest that both molecules are highly dependent on CDR3 length and sequence variation to form specific contacts with antigens. This suggests a general "logic' behind TCR and Ig genetics as it relates to structure and function that helps to explain certain anomalous findings and makes a number of clear predictions.

  View details for Web of Science ID A1997BH80V00009

  View details for PubMedID 9107414

• Biological and chemical aspects of T-cell receptor-mediated recognition 40th Symposium of the Alfred-Benzon-Foundation on HLA and Disease - the Molecular Basis DAVIS, M. M., Lyons, D. S., Altman, J. D., Hampl, J., BONIFACE, J. J., Arden, B., Chien, Y. MUNKSGAARD. 1997: 193–203

  View details for Web of Science ID A1997BJ08M00014

• Ligand recognition by gamma delta T cells 40th Symposium of the Alfred-Benzon-Foundation on HLA and Disease - the Molecular Basis Chien, Y. H., SCHILD, H., Mavadatt, N., Rock, E., Davis, M. MUNKSGAARD. 1997: 179–192

  View details for Web of Science ID A1997BJ08M00013

• Early biochemical signals arise from low affinity TCR-ligand reactions at the cell-cell interface JOURNAL OF EXPERIMENTAL MEDICINE Beeson, C., Rabinowitz, J., Tate, K., Gutgemann, I., Chien, Y. H., Jones, P. P., DAVIS, M. M., McConnell, H. M. 1996; 184 (2): 777-782

  Abstract

  The kinetics of acid release by a mixture of T cells and antigen presenting cells were measured with a microphysiometer during a brief exposure to antigenic peptides. We find that some of the early biochemical events that lead to cellular proliferation cause a specific increase in the rate of acid release. The duration of this increase in acid release reflects the life-time of the peptide-MHC complexes. Peptides that form long-lived complexes produce a response that is stable for more than an hour. Serial TCR engagement is suggested by the observation that the amplitude of this stable response can be rapidly shifted up or down with additional agonist peptide or with antibodies that block T cell receptor binding. Cells briefly exposed to a peptide that forms short-lived peptide-MHC complexes produce a response that decays rapidly as peptide is washed away. A quantitative analysis of the kinetics of this decay in acidification demonstrates that intercellular TCR-ligand reactions are rapid, reversible, and of low apparent affinity with < 20% of peptide-MHC ligand bound to a TCR at any one time. These results demonstrate that the fraction of peptide-MHC ligands bound to TCRs at the cell-cell interface is no higher than anticipated from the affinities observed in solution for isolated TCRs and ligands.

  View details for Web of Science ID A1996VC33700052

  View details for PubMedID 8760833

• A TCR binds to antagonist ligands with lower affinities and faster dissociation rates than to agonists IMMUNITY Lyons, D. S., Lieberman, S. A., Hampl, J., BONIFACE, J. J., Chien, Y. H., Berg, L. J., DAVIS, M. M. 1996; 5 (1): 53-61

  Abstract

  T lymphocyte activation is mediated by the interaction of specific TCR with antigenic peptides bound to MHC molecules. Single amino acid substitutions are often capable of changing the effect of a peptide from stimulatory to antagonistic. Using surface plasmon resonance, we have analyzed the interaction between a complex consisting of variants of the MCC peptide bound to a mouse class II MHC (Ek) and a specific TCR. Using both an improved direct binding method as well as a novel inhibition assay, we show that the affinities of three different antagonist peptide-Ek complexes are approximately 10-50 times lower than that of the wildtype MCC-Ek complex for the TCR, largely due to an increased off-rate. These results suggest that the biological effects of peptide antagonists and partial agonists may be largely based on kinetic parameters.

  View details for Web of Science ID A1996UZ45400006

  View details for PubMedID 8758894

• Recognition by gamma/delta T cells ANNUAL REVIEW OF IMMUNOLOGY Chien, Y. H., Jores, R., Crowley, M. P. 1996; 14: 511-532

  Abstract

  In contrast with the study of alpha beta T cells, that of gamma delta T cells is relatively recent and stems from the discovery of their rearranged genes, rather than from any knowledge of their biological function. Thus, experiments designed to characterize their specificity and function have drawn heavily on our knowledge of alpha beta T cells. During the past few years, many studies, especially with mice lacking either alpha beta or gamma delta T cells, have demonstrated that gamma delta T cells can contribute to immune competence, but they do so in a way that is distinct from alpha beta T cells. It is also evident that gamma delta T cells may not recognize antigen the same way as do alpha beta T cells. Analysis of three protein antigens-the murine MHC class II IEk, the nonclassical MHC T10/T22, and the Herpes virus glycoprotein gI-indicates that gamma delta T cell recognition does not require antigen processing and that the proteins are recognized directly. In all three cases, recognition by these T cell clones involves neither peptides bound to these proteins nor peptides derived from them. Moreover, a group of small phosphate-containing nonpeptide compounds derived from mycobacterial extracts has been found to stimulate a major population of human peripheral gamma delta T cells in a T cell receptor (TCR)-dependent manner. This indicates that gamma delta T cells can respond to ligands that are different from those of alpha beta T cells. Analysis of complementarity determining region (CDR3) length distributions of gamma and delta chains indicates that they are more similar to those of immunoglobulins than to TCR alpha and beta. This further supports the idea that gamma delta and alpha beta T cells recognize antigens differently and suggests that gamma delta T cells may be more like immunoglobulins in their recognition properties. gamma delta T cells share many cell surface proteins with alpha beta T cells and are able to secrete lymphokines and express cytolytic activities in response to antigenic stimulation. These, together with the results cited above, indicate that gamma delta T cells can mediate cellular immune functions without a requirement for antigen processing. Thus, pathogens, damaged tissues, or even B and T cells can be recognized directly, and cellular immune responses can be initiated without a requirement for antigen degradation or specialized antigen-presenting cells. This would give gamma delta T cells greater flexibility than the more classical type of alpha beta T cell-mediated cellular immunity.

  View details for Web of Science ID A1996UH42900021

  View details for PubMedID 8717523

• Ligand recognition by gamma delta T cells 8th International Congress of Mucosal Immunology Chien, Y. H. ACADEMIC PRESS INC. 1996: 169–182

  View details for Web of Science ID A1996BG31B00014

• GAMMA-DELTA T-CELLS - T-CELLS WITH B-CELL-LIKE RECOGNITION PROPERTIES CURRENT BIOLOGY Chien, Y. H., Jores, R. 1995; 5 (10): 1116-1118

  Abstract

  Recent results suggest there are fundamental differences in antigen recognition by alpha beta and gamma delta T-cell receptors, which might underlie possible differences in function between these two types of T cell.

  View details for Web of Science ID A1995TA46400011

  View details for PubMedID 8548281

• FUNCTIONAL EXPRESSION AND RECOGNITION OF NONCLASSICAL MHC CLASS-I T10(B) IS NOT PEPTIDE-DEPENDENT JOURNAL OF IMMUNOLOGY Kaliyaperumal, A., Falchetto, R., Cox, A., Dick, R., Shabanowitz, J., Chien, Y. H., Matis, L., Hunt, D. F., Bluestone, J. A. 1995; 155 (5): 2379-2386

  Abstract

  Studies of classical and nonclassical MHC class I molecules have shown that unique peptides are associated and functionally recognized by alloreactive T cells. We have recently shown that an alloreactive TCR-gamma delta cell recognizes a nonclassical MHC molecule, T10b. However, T cell recognition of this glycoprotein did not appear to require typical peptide recognition based on studies using transporter-defective mutant cell lines. In the current study, we have analyzed in detail, the role of peptide in T10b expression and recognition. The findings reveal that the recognition of the nonclassical MHC molecule by TCR-gamma delta cells is independent of species, tissue type, both the class I and class II Ag processing and presentation pathways, or the presence of peptides. In fact, biochemical analysis of the T10b chimeric molecule, T10b/Ld, transfected into CHO cells using radiolabeled [3H]leucine, HPLC, and mass spectrometry suggest that peptides are not associated with this nonclassical class I molecule. Therefore, some class I molecules, e.g., T10b, do not associate with polymorphic peptides typical of classical MHC class I molecules and can be expressed in the absence of peptides on the cell surface in a functionally active form.

  View details for Web of Science ID A1995RP74300010

  View details for PubMedID 7650372

• ISSUES CONCERNING THE NATURE OF ANTIGEN RECOGNITION BY ALPHA-BETA AND GAMMA-DELTA T-CELL RECEPTORS 9th International Congress of Immunology DAVIS, M. M., Chien, Y. H. ELSEVIER SCI LTD. 1995: 316–18

  View details for Web of Science ID A1995RG98300006

  View details for PubMedID 7576062

• T-CELL RECEPTOR V-REGION USAGE AND ANTIGEN-SPECIFICITY - THE CYTOCHROME-C MODEL SYSTEM Conference on T-Cell Receptor Use in Human Autoimmune Diseases DAVIS, M. M., MCHEYZERWILLIAMS, M., Chien, Y. H. NEW YORK ACAD SCIENCES. 1995: 1–11

  Abstract

  Investigations of the I-Ek-restricted, cytochrome c-specific T-cell response in mice show that both T-cell receptor V alpha and V beta CDR3 residues and the use of particular V alpha s and V beta s are necessary for recognition. Data strongly suggest that specific CDR3 residues are important in contacting the peptide. Other experiments indicate that the requirement for V alpha:V beta conservation is not the result of strong TCR-->MHC interactions, as no correlation was found between V beta usage and changes in the alpha-helixes of the I-Ek molecule. It is also apparent that changes in V alpha or V beta usage could be elicited by changes in the side chain size of single amino acids of the antigenic peptides, suggesting that V alpha or V beta conservation is important for peptide recognition, either directly or indirectly. We also show that we can follow the cytochrome c response in vivo even in nontransgenic mice, solely by staining with anti-V region antibodies as well as mAbs directed at the activation markers CD44 and L-selectin.

  View details for Web of Science ID A1995BD46M00001

  View details for PubMedID 7645810

• The recognition of MHC molecules by gamma delta T cells. Behring Institute Mitteilungen SCHILD, H., Chien, Y. H. 1994: 113-123

  View details for PubMedID 7998905

• THE NATURE OF MAJOR HISTOCOMPATIBILITY COMPLEX RECOGNITION BY GAMMA-DELTA-T-CELLS CELL SCHILD, H., Mavaddat, N., Litzenberger, C., Ehrich, E. W., DAVIS, M. M., Bluestone, J. A., Matis, L., Draper, R. K., Chien, Y. H. 1994; 76 (1): 29-37

  Abstract

  Despite intensive efforts, the general rules for gamma delta T cell recognition remain undefined. Here, we take advantage of the detailed knowledge of the molecular structure and biosynthetic pathways of major histocompatibility complex (MHC) molecules to analyze the recognition properties of the gamma delta T cell clones LBK5 (specific for the class II MHC, IEk) and G8 (specific for the nonclassical class I MHC, TL10b). We find that the activation of these clones requires neither class I nor class II antigen-processing and that peptides do not confer specificity. Epitope mapping also shows that the topology of gamma delta T cell receptor interaction with the MHC is distinct from that of alpha beta T cells. These results suggest that the molecular nature of gamma delta T cell recognition is fundamentally different than that of alpha beta T cells.

  View details for Web of Science ID A1994MR49500004

  View details for PubMedID 8287478

• CDR3 LENGTH IN ANTIGEN-SPECIFIC IMMUNE RECEPTORS JOURNAL OF EXPERIMENTAL MEDICINE ROCK, E. P., SIBBALD, P. R., DAVIS, M. M., Chien, Y. H. 1994; 179 (1): 323-328

  Abstract

  In both immunoglobulins (Ig) and T cell receptors (TCR), the rearrangement of V, D, and J region sequence elements during lymphocyte maturation creates an enormous degree of diversity in an area referred to as the complementarity determining region 3 (CDR3) loop. Variations in the particular V, D, and J elements used, precise points of recombination, and random nucleotide addition all lead to extensive length and sequence heterogeneity. CDR3 loops are often critical for antigen binding in Igs and appear to provide the principal peptide binding residues in TCRs. To better understand the physical and selective constraints on these sequences, we have compiled information on CDR3 size variation for Ig H, L (kappa and lambda) and TCR alpha, beta, gamma, and delta. Ig H and TCR delta CDR3s are the most variable in size and are significantly longer than L and gamma chains, respectively. In contrast, TCR alpha and beta chain distributions are highly constrained, with nearly identical average CDR3 lengths, and their length distributions are not altered by thymic selection. Perhaps most significantly, these CDR3 length profiles suggest that gamma/delta TCRs are more similar to Igs than to alpha/beta TCRs in their putative ligand binding region, and thus gamma/delta and alpha/beta T cells may have fundamentally different recognition properties.

  View details for Web of Science ID A1994MP51700033

  View details for PubMedID 8270877

• HOW ALPHA-BETA-T-CELL RECEPTORS SEE PEPTIDE MHC COMPLEXES IMMUNOLOGY TODAY Chien, Y. H., DAVIS, M. M. 1993; 14 (12): 597-602

  Abstract

  Recent results have added new information to our understanding of alpha beta T-cell receptor mediated recognition. In particular, we find that the V(D)J junction or 'CDR3' portion of TCR alpha and beta seem most important in contacting peptides bound to MHC molecules, consistent with previous predictions. Surprisingly, these same CDR3-peptide contacts also appear to have a major influence on the TCR-MHC molecule interactions as well.

  View details for Web of Science ID A1993MK79800005

  View details for PubMedID 8305132

• T-CELL RECEPTOR INTERACTION WITH PEPTIDE MAJOR HISTOCOMPATIBILITY COMPLEX (MHC) AND SUPERANTIGEN MHC LIGANDS IS DOMINATED BY ANTIGEN JOURNAL OF EXPERIMENTAL MEDICINE Ehrich, E. W., DEVAUX, B., ROCK, E. P., JORGENSEN, J. L., DAVIS, M. M., Chien, Y. H. 1993; 178 (2): 713-722

  Abstract

  While recent evidence strongly suggests that the third complementarity determining regions (CDR3s) of T cell receptors (TCRs) directly contact antigenic peptides bound to major histocompatibility complex (MHC) molecules, the nature of other TCR contact(s) is less clear. Here we probe the extent to which different antigens can affect this interaction by comparing the responses of T cells bearing structurally related TCRs to cytochrome c peptides and staphylococcal enterotoxin A (SEA) presented by 13 mutant antigen-presenting cell (APC) lines. Each APC expresses a class II MHC molecule (I-Ek) with a single substitution of an amino acid residue predicted to be located on the MHC alpha helices and to point "up" towards the TCR. We find that very limited changes (even a single amino acid) in either a CDR3 loop of the TCR or in a contact residue of the antigenic peptide can have a profound effect on relatively distant TCR/MHC interactions. The extent of these effects can be as great as that observed between T cells bearing entirely different TCRs and recognizing different peptides. We also find that superantigen presentation entails a distinct mode of TCR/MHC interaction compared with peptide presentation. These data suggest that TCR/MHC contacts can be made in a variety of ways between the same TCR and MHC, with the final configuration apparently dominated by the antigen. These observations suggest a molecular basis for recent reports in which either peptide analogues or superantigens trigger distinct pathways of T cell activation.

  View details for Web of Science ID A1993LP83000037

  View details for PubMedID 8393480

• TOPOLOGY AND AFFINITY OF T-CELL RECEPTOR MEDIATED RECOGNITION OF PEPTIDE MHC COMPLEXES CURRENT OPINION IN IMMUNOLOGY DAVIS, M. M., Chien, Y. H. 1993; 5 (1): 45-49

  Abstract

  Significant progress has been made on several long-standing issues regarding T-cell receptor mediated recognition of antigen-MHC complexes. For one, early data suggest that the affinity of the T-cell receptor for the peptide-MHC complex is extremely low, with a KD of approximately 10(-4)-10(-5)M, much weaker than most antibody-antigen interactions. The fact that this affinity is lower than that of some T-cell adhesion molecules for their ligands could have important implications for immune surveillance. A second area of interest is the topology of T-cell receptor recognition; evidence of direct contact between the third complementarity determining region of the T-cell receptor and peptide determinants has been obtained. In addition, the orientation of the T-cell receptor with respect to several antigen-MHC complexes has been predicted. They suggest that whereas most or all peptides seem to bind in the same orientation in both class I and class II MHC molecules, the orientation of the T-cell receptor over the peptide-MHC complex may not be fixed.

  View details for Web of Science ID A1993KN29300008

  View details for PubMedID 8452673

• DEVELOPMENT OF GAMMA-DELTA T-CELL SUBSETS FROM FETAL HEMATOPOIETIC STEM-CELLS ANNALS OF THE NEW YORK ACADEMY OF SCIENCES Ikuta, K., Kina, T., MacNeil, I., Uchida, N., Peault, B., Chien, Y. H., Weissman, I. L. 1992; 651: 21-32

  Abstract

  Hematopoietic stem cells (HSC) were isolated from mouse fetus, and their developmental potential was compared with adult HSC. Donor-derived V gamma 3+T cells were detected in fetal thymic lobes, repopulated in vitro with fetal liver HSC, but not in those with adult bone marrow HSC. Single clonogenic fetal HSC gave rise to thymic progeny that include V gamma 3+, other gamma delta+, and alpha beta+ T cells. No V gamma 3+ T cells were detected in adult thymus injected intrathymically with either fetal or adult HSC. These results support a hypothesis that only fetal HSC have the capacity to differentiate into V gamma 3+ T cells in the fetal thymic microenvironment, and that the developmental potential of HSC may change during ontogeny.

  View details for Web of Science ID A1992JM21900003

  View details for PubMedID 1599125

• DEVELOPMENT OF GAMMA-DELTA T-CELL SUBSETS FROM FETAL HEMATOPOIETIC STEM-CELLS CONF ON CD5 B-CELLS IN DEVELOPMENT AND DISEASE Ikuta, K., Kina, T., MacNeil, I., Uchida, N., Peault, B., Chien, Y. H., Weissman, I. L. NEW YORK ACAD SCIENCES. 1992: 21–32

  View details for Web of Science ID A1992BW41S00003

• A GAMMA-DELTA T-CELL RECEPTOR HETERODIMER INDUCES THE EXPRESSION OF CD4 AND CD8 IN THYMOCYTES JOURNAL OF EXPERIMENTAL MEDICINE Iwashima, M., DAVIS, M. M., Chien, Y. H. 1991; 174 (1): 293-296

  Abstract

  CD4 and CD8 have been useful surface markers for alpha/beta T cell maturation. In an alpha/beta T cell receptor (TCR) transgenic SCID mice system, it has been shown that alpha/beta TCR alone is sufficient to induce CD4 and CD8 surface expression on thymic T cells. Although the late embryonic thymic gamma/delta T cells are predominately single and double positive, it has not been clear if gamma/delta TCR has a similar capacity. In this study, we show that when transgenes encoding the earliest embryonic gamma/delta TCR are coexpressed with the SCID defect, the gamma/delta transgenes promote the appearance of both the CD4-8- and CD4+8+ T cells in the thymus. Furthermore, the expression of CD4 and CD8 does not require continuous surface gamma/delta TCR expression. These results indicate that gamma/delta TCR alone can promote the CD4/8 surface expression, and may suggest a role for gamma/delta T cells in initiating normal thymic ontogeny.

  View details for Web of Science ID A1991FU89700034

  View details for PubMedID 1905341

• RECOGNITION OF SELF ANTIGENS BY SKIN-DERIVED T-CELLS WITH INVARIANT GAMMA-DELTA-ANTIGEN RECEPTORS SCIENCE Havran, W. L., Chien, Y. H., Allison, J. P. 1991; 252 (5011): 1430-1432

  Abstract

  Thy-1+ dendritic epidermal T cells (dECs) express invariant gamma delta antigen receptors and are found in intimate contact with keratinocytes in murine epidermis--thus raising the possibility that keratinocytes express a ligand for the antigen receptor of these T cells. Thy-1+ dECs were stimulated to produce lymphokines by interaction with keratinocytes in vitro. This stimulation was mediated through the dEC antigen receptor and did not appear to be restricted by the major histocompatibility complex. Thus, dECs can recognize self antigens and may participate in immune surveillance for cellular damage rather than for foreign antigens.

  View details for Web of Science ID A1991FP86800036

  View details for PubMedID 1828619

• EXPRESSION OF A FETAL GAMMA-DELTA T-CELL RECEPTOR IN ADULT MICE TRIGGERS A NON-MHC-LINKED FORM OF SELECTIVE DEPLETION INTERNATIONAL IMMUNOLOGY Iwashima, M., Green, A., Bonyhadi, M., DAVIS, M. M., Allison, J. P., Chien, Y. 1991; 3 (4): 385-393

  Abstract

  Day 14 fetal thymocytes and adult dendritic epidermal T cells (dEC) of all mouse strains express a characteristic non-polymorphic gamma delta T-cell receptor which is rarely found in the adult thymus or lymph nodes. We have made transgenic mice expressing this particular set of receptors on T cells in C3H and C57BL/6 mice. In adult mice of the latter strain, a dramatic depletion of transgene expressing T cells occurs and this effect is primarily mediated by thymic radiosensitive cells. The depletion is genetically dominant but not MHC-linked with major factor(s) mapping to chromosome 18. Taken together, our results show that strain-specific developmental changes in the thymic environment may play a role in shaping the gamma delta TCR repertoire.

  View details for Web of Science ID A1991FK38000012

  View details for PubMedID 1831656

• A DEVELOPMENTAL SWITCH IN THYMIC LYMPHOCYTE MATURATION POTENTIAL OCCURS AT THE LEVEL OF HEMATOPOIETIC STEM-CELLS CELL Ikuta, K., Kina, T., MacNeil, I., Uchida, N., Peault, B., Chien, Y. H., Weissman, I. L. 1990; 62 (5): 863-874

  Abstract

  Hematopoietic stem cells (HSCs) isolated from mouse fetal liver, like adult HSCs, are Thy-1lo Lin- Sca-1+. Donor-derived V gamma 3+ T cells were detected in fetal thymic lobes repopulated in vitro with fetal liver HSCs, but not in those with adult bone marrow HSCs. Single clonogenic fetal HSCs gave rise to thymic progeny that include V gamma 3+, other gamma delta+, and alpha beta+ T cells. No V gamma 3+ T cells were detected in adult thymus injected intrathymically with either fetal or adult HSCs. These results support the hypothesis that only fetal HSCs have the capacity to differentiate into V gamma 3+ T cells in the fetal thymic microenvironment and that the developmental potential of HSCs may change during ontogeny.

  View details for Web of Science ID A1990DY10000006

  View details for PubMedID 1975515

• A POSSIBLE BASIS FOR MAJOR HISTOCOMPATIBILITY COMPLEX-RESTRICTED T-CELL RECOGNITION PHILOSOPHICAL TRANSACTIONS OF THE ROYAL SOCIETY OF LONDON SERIES B-BIOLOGICAL SCIENCES DAVIS, M. M., Chien, Y. H., BJORKMAN, P. J., ELLIOTT, J. F., Iwashima, M., ROCK, E. P., PATTEN, P. A. 1989; 323 (1217): 521-524

  Abstract

  Four distinct T-cell antigen-receptor gene loci have now been identified and partly characterized: alpha, beta, gamma and delta. All of these loci can rearrange in an immunoglobulin-like fashion and express polypeptides that contribute to either alpha:beta or gamma:delta T-cell receptor-CD3 complexes. Surprisingly, the T-cell receptor (TCR) delta coding regions are located entirely, or almost entirely, within the TCR alpha locus and share at least some of the V region gene segments, thus at least partly linking the two different types of receptor heterodimers. Analysis of potential T-cell receptor diversity, particularly that of the delta chain, indicates a striking concentration of somatic polymorphism in the V-J junctional region of the two heterodimers, four to six orders of magnitude higher than similar calculations for immunoglobulin light- and heavy-chain combinations. In contrast, the number of possible V region combinations in T-cell receptors is one hundredth to one thousandth that of immunoglobulins. TCR alpha: beta heterodimers are known to recognize many possible fragments of antigens embedded in the peptide-binding clefts of a relatively small number of major histocompatibility complex (MHC) molecules. Thus it is attractive to speculate that the V-J junctional portions of both types of T-cell receptor contact peptide antigens, whereas the remaining diversity regions contact the MHC. This contention is supported by molecular modelling studies and has interesting implications for the evolution of antigen-receptor genes.

  View details for Web of Science ID A1989AB41200008

  View details for PubMedID 2569209

• VARIABLE REGION (V-SIGMA) GENE SEGMENT MOST FREQUENTLY UTILIZED IN ADULT THYMOCYTES IS 3' OF THE CONSTANT (C-SIGMA) REGION PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA Iwashima, M., Green, A., DAVIS, M. M., Chien, Y. H. 1988; 85 (21): 8161-8165

  Abstract

  A variable region (V delta 5) of the T-cell receptor (TcR) delta chain that is preferentially expressed in adult murine thymocytes is located 2.5 kilobases 3' of the constant region (C delta) element. The V delta 5 coding sequence is in a transcriptional orientation opposite the J delta (joining region) and C delta coding elements and rearranges by inversion. The C delta is divided into four exons, three of which encode amino acids of TcR delta polypeptide, and the fourth comprises the entire 3' untranslated region. In this respect, C delta resembles C alpha rather than C beta or C gamma.

  View details for Web of Science ID A1988Q834100070

  View details for PubMedID 3263646

• THE ADULT T-CELL RECEPTOR DELTA-CHAIN IS DIVERSE AND DISTINCT FROM THAT OF FETAL THYMOCYTES NATURE ELLIOTT, J. F., ROCK, E. P., PATTEN, P. A., DAVIS, M. M., Chien, Y. H. 1988; 331 (6157): 627-631

  Abstract

  T lymphocytes recognize foreign molecules using the T-cell receptor (TCR), a disulphide-linked heterodimer closely associated with the CD3 polypeptide complex on the cell surface. The TCR alpha beta heterodimers seem largely responsible for the recognition properties of both helper (TH) and cytotoxic (TC) T cells. Recently, a second CD3-associated T-cell receptor heterodimer, gamma delta, has been described. Cells bearing the gamma delta receptor appear before those bearing alpha beta during thymic ontogeny and persist as a minor component (1-10%) of mature peripheral T cells. Their function is unknown. As there are a limited number of functional TCR V gamma gene segments, the size and potential diversity of the V delta repertoire is important for the number of different antigens that may be recognized by gamma delta heterodimers. The delta-chain locus is located 75 kilobases (kb) 5' to the TCR C alpha coding region, raising the possibility that the alpha and delta V-region repertoires may overlap. Also, analysis of rearrangements at the delta-chain locus in developing thymocytes shows distinct fetal and adult patterns indicating that there may be differences between the fetal and adult V delta repertoires. To address these questions, we have characterized a large number of delta-containing complementary DNA clones from adult double-negative thymocytes (CD4-8-), an immature population that is enriched for gamma delta-bearing cells. We find that a limited number of V delta sequences are used, showing little overlap with known adult V alpha s and differing significantly from fetal V delta s. But as two D elements may participate simultaneously in V delta gene assembly, and random nucleotides may be added at any one of three junctional points, the potential number of different delta chains that can be made in the adult thymus is very large (approximately 10(13)).

  View details for Web of Science ID A1988M118000061

  View details for PubMedID 2963227

• T-CELL RECEPTOR DELTA-GENE REARRANGEMENTS IN EARLY THYMOCYTES NATURE Chien, Y. H., Iwashima, M., Wettstein, D. A., Kaplan, K. B., ELLIOTT, J. F., Born, W., DAVIS, M. M. 1987; 330 (6150): 722-727

  Abstract

  The T-cell receptor delta-chain variable region can be assembled from as many as four distinct gene segments, V, D1, D2 and J, more than any other antigen-receptor gene. In fetal thymocytes V----D joinings are as common as D----J or VDJ rearrangements and one V gene segment predominates. Analysis of rearrangements at TCR gamma and delta loci during fetal ontogeny suggests abrupt changes and possible coordinate control in the rearrangement and expression of these loci.

  View details for PubMedID 2961997

• IDENTIFICATION AND SEQUENCE OF A 4TH HUMAN T-CELL ANTIGEN RECEPTOR CHAIN NATURE Loh, E. Y., Lanier, L. L., Turck, C. W., Littman, D. R., DAVIS, M. M., Chien, Y. H., WEISS, A. 1987; 330 (6148): 569-572

  Abstract

  Thymus-derived lymphocytes (T cells) use clonally distributed antigen receptors to recognize peptide fragments associated with products of the major histocompatibility complex (MHC) (refs 1-4). On most murine and human T cells the T cell receptor (TCR) is composed of disulphide-linked alpha and beta chains (TCR alpha/beta), each of which contains constant and variable domains, and which are associated with the invariant chains of the CD3 complex. It has been demonstrated, however, that a distinct CD3-associated TCR is expressed on a small subset of T cells or immature thymocytes which fail to express either CD4 or CD8 (refs 7-14), the molecules associated with class II or class I MHC antigen recognition. Instead of TCR alpha/beta, these cells express heterodimers of gamma and delta chains (TRC gamma/delta). The genes encoding alpha, beta, and gamma have been isolated and characterized. A new murine T cell receptor (Cx) gene which undergoes rearrangement and expression early during T cell ontogeny has recently been identified 5' of the murine J alpha C alpha gene locus. Here we isolate and sequence the homologous transcript from PEER, a human cell line that expresses a TCR gamma/delta, and show that it encodes a protein with characteristic V, D, J, and C segments. Using probes derived from this transcript, we have shown that both PEER and MOLT-13, another TCR gamma/delta-expressing cell line, rearrange this locus and express two sizes of transcripts differing in the 3' untranslated region. Using a synthetic peptide derived from the deduced C region sequence, we have prepared antisera that precipitates the delta chain of the TCR from both PEER and MOLT-13, thus demonstrating that Cx and its human homologue code for the delta chain of the TCR.

  View details for Web of Science ID A1987L184100062

  View details for PubMedID 2825032

• TRANSIENT REARRANGEMENTS OF THE T-CELL ANTIGEN RECEPTOR ALPHA-LOCUS IN EARLY THYMOCYTES JOURNAL OF EXPERIMENTAL MEDICINE LINDSTEN, T., FOWLKES, B. J., Samelson, L. E., DAVIS, M. M., Chien, Y. H. 1987; 166 (3): 761-775

  Abstract

  The dull Ly-1 double-negative (Ly-1dull, Lyt-2-, L3T4-) subpopulation appears to be the major precursor group of T lymphocytes in the thymus. In examining the status of the alpha, beta, and gamma chain genes for T cell receptors (TCR) in this population of cells and hybridomas made from them, we find that all of these loci appear to begin DNA rearrangements in a nearly simultaneous fashion. In the case of the gamma genes, these involve V gamma----J gamma C gamma gene rearrangements; with the beta chain genes, both D beta----J beta C beta rearrangement and V beta----D beta J beta C beta rearrangements are evident; and in the case of the alpha locus, assayed in part by pulsed-field gel electrophoresis, they take the form of a novel series of rearrangements occurring 80 kb or more 5' to the C alpha gene. These alpha locus rearrangements are well away from any of the J alpha gene segments found in cDNA clones to date and are deleted in most mature thymocytes and functional T cell lines. Therefore they appear to represent a distinct class of rearrangement that occurs before V alpha----J alpha joining. These distinctions between the character of the TCR gene rearrangements in these cells represent useful markers in further distinguishing different stages of T cell differentiation within this compartment of early T cells. In addition, the unexpected discovery of clonal rearrangements so far away from any of the expressed J alpha gene segments, and at a stage where there is little or no stable C alpha RNA present, has interesting implications for the hierarchy of TCR gene expression.

  View details for Web of Science ID A1987J927400012

  View details for PubMedID 3040885

  View details for PubMedCentralID PMC2188689

• A NEW T-CELL RECEPTOR GENE LOCATED WITHIN THE ALPHA-LOCUS AND EXPRESSED EARLY IN T-CELL DIFFERENTIATION NATURE Chien, Y. H., Iwashima, M., Kaplan, K. B., ELLIOTT, J. F., DAVIS, M. M. 1987; 327 (6124): 677-682

  Abstract

  A new T-cell receptor gene lies just 5' to the J alpha C alpha coding regions. Its placement in this location suggests a novel mechanism for the regulation of expression of one T-cell receptor polypeptide to another during ontogeny. Rearrangement of this locus occurs very early in thymic differentiation and its RNA expression parallels that of the gamma-chain in thymic subpopulations, making this a possible candidate for the recently described delta-chain of the T-cell receptor.

  View details for Web of Science ID A1987H882700046

  View details for PubMedID 2439914

• T-CELL RECEPTOR GENE STRUCTURE AND FUNCTION CELLULAR IMMUNOLOGY DAVIS, M. M., LINDSTEN, T., Gascoigne, N. R., Goodnow, C., Chien, Y. H. 1986; 99 (1): 24-28

  Abstract

  Recent progress in the serology, biochemistry, and now the molecular genetics of T-cell receptor molecules has brought within reach the prospect of solving some of the most basic questions about the nature of T-cell recognition. These include the exact nature of the receptor-major histocompatibility complex (MHC)-antigen recognition event and the sequential expression of T-cell receptor molecules in the thymus.

  View details for Web of Science ID A1986C016400005

  View details for PubMedID 3489561

• PARADIGM REGAINED - SIMILARITIES AND DIFFERENCES BETWEEN T-CELL RECEPTOR AND IMMUNOGLOBULIN GENES BIOCHEMICAL SOCIETY SYMPOSIA DAVIS, M. M., LINDSTEN, T., Gascoigne, N. R., Goodnow, C., Chien, Y. H. 1986: 205-209

  View details for Web of Science ID A1986D410900019

  View details for PubMedID 3101698

• VARIABILITY AND REPERTOIRE SIZE OF T-CELL RECEPTOR V-ALPHA GENE SEGMENTS NATURE Becker, D. M., Patten, P., Chien, Y. H., Yokota, T., ESHHAR, Z., Giedlin, M., Gascoigne, N. R., Goodnow, C., Wolf, R., Arai, K., DAVIS, M. M. 1985; 317 (6036): 430-434

  Abstract

  The immune system of higher organisms is composed largely of two distinct cell types, B lymphocytes and T lymphocytes, each of which is independently capable of recognizing an enormous number of distinct entities through their antigen receptors; surface immunoglobulin in the case of the former, and the T-cell receptor (TCR) in the case of the latter. In both cell types, the genes encoding the antigen receptors consist of multiple gene segments which recombine during maturation to produce many possible peptides. One striking difference between B- and T-cell recognition that has not yet been resolved by the structural data is the fact that T cells generally require a major histocompatibility determinant together with an antigen whereas, in most cases, antibodies recognize antigen alone. Recently, we and others have found that a series of TCR V beta gene sequences show conservation of many of the same residues that are conserved between heavy- and light-chain immunoglobulin V regions, and these V beta sequences are predicted to have an immunoglobulin-like secondary structure. To extend these studies, we have isolated and sequenced eight additional alpha-chain complementary cDNA clones and compared them with published sequences. Analyses of these sequences, reported here, indicate that V alpha regions have many of the characteristics of V beta gene segments but differ in that they almost always occur as cross-hybridizing gene families. We conclude that there may be very different selective pressures operating on V alpha and V beta sequences and that the V alpha repertoire may be considerably larger than that of V beta.

  View details for Web of Science ID A1985ARS3100051

  View details for PubMedID 2995827

• GENOMIC ORGANIZATION AND SEQUENCE OF T-CELL RECEPTOR BETA-CHAIN CONSTANT-REGION AND JOINING-REGION GENES NATURE Gascoigne, N. R., Chien, Y. H., Becker, D. M., Kavaler, J., DAVIS, M. M. 1984; 310 (5976): 387-391

  Abstract

  The genomic structure of the joining (J) and constant (C) regions of the locus encoding the beta-chain of the murine T-cell receptor has been analysed. The gene segments are arranged tandemly (J-C/J-C) within a 15-kilobase region. The two constant-region genes are almost identical, differing by only four amino acids, all in carboxy-terminal portions. Each C region comprises four exons encoding an external globular domain, a small hinge-like region, a transmembrane region and a cytoplasmic tail plus 3'-untranslated region. The two clusters of J regions each contain 7 distinct elements, 12 of which may be functional.

  View details for Web of Science ID A1984TC54400034

  View details for PubMedID 6336329

• A 3RD-TYPE OF MURINE T-CELL RECEPTOR GENE NATURE Chien, Y., Becker, D. M., LINDSTEN, T., Okamura, M., Cohen, D. I., DAVIS, M. M. 1984; 312 (5989): 31-35

  Abstract

  By subtractive cDNA hybridizations, we have isolated a new species of T-cell receptor cDNA clone whose predicted amino acid sequence has homology to variable, constant, joining and diversity segments of immunoglobulins and T-cell receptors. The corresponding genomic sequence is also rearranged in several T-cell DNAs. The four potential N-linked glycosylation sites, frequency of expression and predicted molecular weight (27,800) of this molecule make it a likely candidate for the alpha-chain of the T-cell receptor. Expression data also indicate that this gene may be activated at a later stage of T-cell differentiation than the beta-chain.

  View details for Web of Science ID A1984TQ42600035

  View details for PubMedID 6548551

• A MURINE T-CELL RECEPTOR GENE-COMPLEX - ISOLATION, STRUCTURE AND REARRANGEMENT IMMUNOLOGICAL REVIEWS DAVIS, M. M., Chien, Y., Gascoigne, N. R., HEDRICK, S. M. 1984; 81: 235-258

  Abstract

  We describe here our efforts to develop a systematic strategy for the enumeration and isolation of genes encoding T cell-specific, membrane-associated polypeptides. Of particular importance among the cDNA clones that we have isolated is one which encodes the beta chain of the murine T cell receptor for antigen. The gene product is strikingly similar to immunoglobulin in its variable (V), constant (C), joining (J), and diversity (D) - like elements and the ability of the latter three types of element to assort independently during differentiation. The genomic organization of the constant region locus which encodes this molecule consists of two tandemly arrayed units of the form JC/JC, where the J clusters consist of fourteen distinct elements, many of which may be functional. The two constant regions differ by only four amino acids and both are apparently expressed in T cells of the same phenotype, so they probably do not represent isotypes in the classical sense. The sequences flanking the V, D and J elements are also similar to the conserved heptamer and nonomer sequences of immunoglobulins thought to be important in the recombination of the specific gene segments. These similarities indicate both the common evolutionary origins of these loci and the strong selective pressure that must be operative. The T cell receptor locus differs somewhat from immunoglobulins in the large number of possible J region elements and to some extent in the hypervariable-framework regions of the V region. We and others have mapped this gene to chromosome 6 of the mouse, in the same portion of the chromosome but not closely linked the kappa locus. The significance of the predicted protein structure of the beta chain and its support of the one-receptor model of MHC-antigen recognition are discussed.

  View details for Web of Science ID A1984TT81400012

  View details for PubMedID 6096259

• LOCALIZATION OF A T-CELL RECEPTOR DIVERSITY-REGION ELEMENT NATURE Kavaler, J., DAVIS, M. M., Chien, Y. H. 1984; 310 (5976): 421-423

  Abstract

  Recently, complementary DNA clones encoding one chain of the T-cell receptor for antigen have been isolated from both murine and human cell lines. Sequence analysis of these clones indicates that they encode elements analogous to the variable (V), constant (C) and joining (J) regions of immunoglobins and that this corresponds to the beta-chain subunit of the T-cell receptor complex. These genes are rearranged in the genomes of specific T-cell lines and hybrids but not in other cell types. Analysis of the components of one such rearranged gene, 2B4, isolated from the pigeon cytochrome c-specific, H-2E-restricted T helper (TH) hybridoma, and its unrearranged (liver) counterpart, indicate that an 8-nucleotide sequence 3' to the rearranged variable region is not derived from either the germ-line V- or J-region gene segments. As this sequence lies at a similar position to the diversity (D) region in immunoglobulin heavy-chain genes, we postulated the existence of an array of germ-line D-region elements that would contribute significantly to the number of different beta-chain molecules which could be created. Here we describe the localization and sequence of one such D-region element, found approximately 650 nucleotides 5' to the first JT cluster. This element seems to be involved in both functional (V-D-J) and non-functional (D-J) rearrangements. Our observations, combined with previous results, indicate that variable-region formation (V-D-J joining) in the T-cell receptor beta-chain gene follows the 12/23-base pair (bp) rule of rearrangement established for the recombination of immunoglobulin gene segments, but that the organization of the heptamer and nonamer element found surrounding the D region is significantly different.

  View details for Web of Science ID A1984TC54400049

  View details for PubMedID 6611508

• STRUCTURE, EXPRESSION AND DIVERGENCE OF T-CELL RECEPTOR BETA-CHAIN VARIABLE REGIONS NATURE Patten, P., Yokota, T., Rothbard, J., Chien, Y., Arai, K., DAVIS, M. M. 1984; 312 (5989): 40-46

  Abstract

  Analysis of three new T-cell receptor beta-chain variable regions together with those in the literature indicates that they have both remarkable similarities and differences with those of immunoglobulin. Less than 10 V regions appear to predominate in the thymus. V beta sequences are much more heterogeneous at the amino acid level than are immunoglobulin V regions and they appear to diverge between species much more quickly, apparently the result of additional hypervariable regions. Three of these putative new hypervariable regions lie outside of the classical immunoglobulin binding site, an indication that important interactions may be occurring in these regions with polymorphic MHC determinants.

  View details for Web of Science ID A1984TQ42600037

  View details for PubMedID 6092964

• SOMATIC RECOMBINATION IN A MURINE T-CELL RECEPTOR GENE NATURE Chien, Y. H., Gascoigne, N. R., Kavaler, J., Lee, N. E., DAVIS, M. M. 1984; 309 (5966): 322-326

  Abstract

  A putative T-cell receptor gene was isolated from the DNA of the helper hybridoma, 2B4 . Analysis of the sequence components of this rearranged gene indicates the presence of separate variable (V), diversity (D) and joining (J) region elements analogous to those of heavy-chain immunoglobulins. These findings further support the belief that the products of this locus are involved in antigen recognition by T cells. There is no evidence of somatic mutation between the putative germ-line and the expressed variable-region gene.

  View details for Web of Science ID A1984SS83000032

  View details for PubMedID 6328306