This project consists of the creation of important tools for measuring T cell receptor diversity and TCR-mediated signaling, and use of these tools in both patients and experimental models of TCR diversity. The first tool is the use of high-throughput sequencing to measure the diversity of T-cell receptor rearrangements. The strategies and protocol development are ongoing and being done in collaboration with Danny Douek's lab in the NIH VRC. The next generation-based TCR sequencing method and techniques for analysis are completed. We applied this method to the study of patients with rare diseases of leaky severe combined immune deficiency. These patients have mutations in genes known to cause SCID, but either have a profound lymphoproliferative disorder associated with severe dermatitis, elevated IgE and eosinophilia, called Omenn syndrome, or a milder phenotype, associated with milder effects on protein activity, characterized by autoimmunity and granulomatous disease, called hypomorphic SCID. We found that patients with Omenns syndrome due to mutations in recombinase activating gene (Rag) had marked repertoire constriction, substantially smaller CDR3 regions and impaired N-nucleotide addition compared to those with Omenn syndrome due to other causes, suggesting that normal Rag function is critical for T-cell receptor junctional diversity. We also found that while the TCR repertoire diversity in the hypomorphic SCID patients was similar to controls, and while amino acid usage was similar to controls in Omenn Syndrome patients, there were significantly different residues used in the hypomorphic SCID, arguing for a specific set of TCRs which are pathogenic in their disease and not in a disease characterized by even more profound repertoire reduction. The results of this study were published in the Journal of Allergy and Clinical Immunology. Our findings were validated in a companion manuscript in the Journal of Allergy and Clinical Immunology from the Notarangelo lab showing similar results in an in vitro B-cell receptor recombination system, interrogating the various mutations in Rag. We further attempted to interrogate the sequences of patients with graft-vs-host disease soon after transplant to see if they correlated to any sequences found in our patient cohort. No overlap was found.

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7
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2015
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Mitson-Salazar, Alyssa; Yin, Yuzhi; Wansley, Daniel L et al. (2016) Hematopoietic prostaglandin D synthase defines a proeosinophilic pathogenic effector human T(H)2 cell subpopulation with enhanced function. J Allergy Clin Immunol 137:907-18.e9
Li, Hoi Ming; Hiroi, Toyoko; Zhang, Yongqing et al. (2016) TCR? repertoire of CD4+ and CD8+ T cells is distinct in richness, distribution, and CDR3 amino acid composition. J Leukoc Biol 99:505-13
Wan, Chi-Keung; Andraski, Allison B; Spolski, Rosanne et al. (2015) Opposing roles of STAT1 and STAT3 in IL-21 function in CD4+ T cells. Proc Natl Acad Sci U S A 112:9394-9
Knight, John M; Mak, Garbo; Shaw, Joanne et al. (2015) Long-Acting Beta Agonists Enhance Allergic Airway Disease. PLoS One 10:e0142212
Zhang, Yu; Yu, Xiaomin; Ichikawa, Mie et al. (2014) Autosomal recessive phosphoglucomutase 3 (PGM3) mutations link glycosylation defects to atopy, immune deficiency, autoimmunity, and neurocognitive impairment. J Allergy Clin Immunol 133:1400-9, 1409.e1-5
Yu, Xiaomin; Almeida, Jorge R; Darko, Sam et al. (2014) Human syndromes of immunodeficiency and dysregulation are characterized by distinct defects in T-cell receptor repertoire development. J Allergy Clin Immunol 133:1109-15
Barber, John S; Yokomizo, Lauren K; Sheikh, Virginia et al. (2013) Peptide library-based evaluation of T-cell receptor breadth detects defects in global and regulatory activation in human immunologic diseases. Proc Natl Acad Sci U S A 110:8164-9
Lawrence, Monica G; Barber, John S; Sokolic, Robert A et al. (2013) Elevated IgE and atopy in patients treated for early-onset ADA-SCID. J Allergy Clin Immunol 132:1444-6
Paul, William E; Milner, Joshua D; Grossman, Zvi (2013) Pathogen-sensing, regulatory T cells, and responsiveness-tuning collectively regulate foreign- and self-antigen mediated T-cell responses. Cold Spring Harb Symp Quant Biol 78:265-76
Hirahara, Kiyoshi; Poholek, Amanda; Vahedi, Golnaz et al. (2013) Mechanisms underlying helper T-cell plasticity: implications for immune-mediated disease. J Allergy Clin Immunol 131:1276-87

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