Peptide:MHC (pMHC) tetramers are reagents that can identify T cells based directly on their antigen receptor specificity. Recent developments in the generation and use of these reagents have made them a powerful new tool with which to study antigen-specific T cell responses, particularly in humans where experimental manipulation of T cells is extremely limited by feasibility issues. The overall objective of the Tetramer Core is to provide researchers from each of the proposed projects with the resources necessary to use tetramer technology to study allergen-specific T cells in mice and humans.
The specific aims of the Tetramer Core will be 1) to generate pMHCII tetramers to defined allergens studied in the proposed projects, and 2) to provide technical expertise in tetramer-based enrichment techniques to enable identification of low frequency allergen-specific T cells. State-of-the-art tetramers will be generated incorporating the most advanced technical refinements to date. Recombinant DNA constructs encoding soluble forms of pMHCII molecules fused to immunogenic peptide epitopes from defined allergens will be cloned and expressed in an insect cell expression system. Purified biotinylated pMHCII complexes will then be tetramerized via their interaction with fluorescently labeled streptavidin molecules. For each allergen studied in human subjects, a panel of tetramers corresponding to several common HLA-DR alleles will be generated to ensure adequate applicability of these reagents to the HLA-diverse study population. Because antigen-specific T cell populations are often present in very low frequencies, tetramers will often be used in conjunction with antibody-coated magnetic beads to enable enrichment of tetramer-positive cells. This will allow for high-resolution detection of allergen-specific T cells from small amounts of human cell samples. The Tetramer Core will provide technical support to researchers so they can quickly master these techniques. Collectively, the services provided by the Tetramer Core will enable researchers to immediately take advantage of this powerful technology that otherwise may not be readily available at a practical scale for many years.

Public Health Relevance

The use of peptide:MHC tetramers to identify allergen-specific T cells elevates the study of allergic immune responses to a powerful new level. This technology fills a longstanding void in our ability to directly study the T cell component of allergy and asthma, particularly in humans, and should contribute greatly to our ultimate goal of understanding and treating these immune disorders.

Agency
National Institute of Health (NIH)
Type
Research Program--Cooperative Agreements (U19)
Project #
5U19AI095261-04
Application #
8707954
Study Section
Special Emphasis Panel (ZAI1)
Project Start
Project End
Budget Start
Budget End
Support Year
4
Fiscal Year
2014
Total Cost
Indirect Cost
Name
Massachusetts General Hospital
Department
Type
DUNS #
City
Boston
State
MA
Country
United States
Zip Code
02199
Cho, Josalyn L; Ling, Morris F; Adams, David C et al. (2016) Allergic asthma is distinguished by sensitivity of allergen-specific CD4+ T cells and airway structural cells to type 2 inflammation. Sci Transl Med 8:359ra132
Urso, Katia; Alvarez, David; Cremasco, Viviana et al. (2016) IL4RA on lymphatic endothelial cells promotes T cell egress during sclerodermatous graft versus host disease. JCI Insight 1:
Hondowicz, Brian D; An, Dowon; Schenkel, Jason M et al. (2016) Interleukin-2-Dependent Allergen-Specific Tissue-Resident Memory Cells Drive Asthma. Immunity 44:155-66
Proekt, Irina; Miller, Corey N; Jeanne, Marion et al. (2016) LYN- and AIRE-mediated tolerance checkpoint defects synergize to trigger organ-specific autoimmunity. J Clin Invest 126:3758-3771
Jhunjhunwala, Siddharth; Alvarez, David; Aresta-DaSilva, Stephanie et al. (2016) Frontline Science: Splenic progenitors aid in maintaining high neutrophil numbers at sites of sterile chronic inflammation. J Leukoc Biol 100:253-60
Ling, Morris F; Luster, Andrew D (2016) Allergen-Specific CD4(+) T Cells in Human Asthma. Ann Am Thorac Soc 13 Suppl 1:S25-30
Leung, John; Mehrzad, Raman; Hundal, Navneet Virk et al. (2015) Longitudinal Perspective on Managing Refractory Eosinophilic Esophagitis. J Allergy Clin Immunol Pract 3:951-6
Patil, Sarita U; Ogunniyi, Adebola O; Calatroni, Agustin et al. (2015) Peanut oral immunotherapy transiently expands circulating Ara h 2-specific B cells with a homologous repertoire in unrelated subjects. J Allergy Clin Immunol 136:125-134.e12
Shade, Kai-Ting C; Platzer, Barbara; Washburn, Nathaniel et al. (2015) A single glycan on IgE is indispensable for initiation of anaphylaxis. J Exp Med 212:457-67
Mattapallil, Mary J; Silver, Phyllis B; Cortes, Lizette M et al. (2015) Characterization of a New Epitope of IRBP That Induces Moderate to Severe Uveoretinitis in Mice With H-2b Haplotype. Invest Ophthalmol Vis Sci 56:5439-49

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