T cells play a central role in the human immune system and their depletion leads to increased susceptibility to infections and some malignancies. T cell development requires a period of education in the thymus, an organ that involutes in late adolescence. If T cells suitable for use in human patients could be generated in vitro, it could lead to novel immunologic therapies for infections, immunodeficiencies and malignancies. Existing In vitro systems that support T cell development use co-culture with allogeneic human or animal tissues, making them less suitable for the production of T cells for use in humans. There are remarkable similarities between the epithelial and stromal cells of the thymus and keratinocytes and fibroblasts of skin. I present evidence that human skin cells arrayed on a three-dimensional matrix support the full process of ? human T cell development. Newly generated T cells are diverse, functionally mature, and tolerant to ? self-MHC. To extend this work, I propose to 1) determine if monolayer cultures of skin cells can support T cell development, 2) to examine what stages of development are supported by keratinocytes vs. fibroblasts and to determine if skin cells transduced with the Notch ligand delta-like 1 support augmented T cell production, 3) to examine self-tolerance in T cells generated in cultures with skin cells transduced to express AIRE, 4) to demonstrate that fully autologous T cells can be generated using skin and bone marrow from a single individual and 5) to examine the ability of T cells generated in skin cell cultures to reconstitute immunodeficient mice. My goal is to develop a system in which samples of adult skin and bone marrow can be used to generate autologous, self-tolerant T cells for the treatment of human disease. This proposal describes a mentored four-year career development plan designed to facilitate my transition from post-doctoral fellow'to independent investigator. It takes advantage of the many resources available to me in the laboratory of Dr. Thomas Kupper and in the Harvard Medical School environment. ? ?

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Clinical Investigator Award (CIA) (K08)
Project #
5K08AI060890-02
Application #
7172583
Study Section
Allergy & Clinical Immunology-1 (AITC)
Program Officer
Prograis, Lawrence J
Project Start
2006-02-01
Project End
2010-01-31
Budget Start
2007-02-01
Budget End
2008-01-31
Support Year
2
Fiscal Year
2007
Total Cost
$135,000
Indirect Cost
Name
Brigham and Women's Hospital
Department
Type
DUNS #
030811269
City
Boston
State
MA
Country
United States
Zip Code
02115
Clark, Rachael A (2013) Human skin in the game. Sci Transl Med 5:204ps13
Seneschal, Julien; Clark, Rachael A; Gehad, Ahmed et al. (2012) Human epidermal Langerhans cells maintain immune homeostasis in skin by activating skin resident regulatory T cells. Immunity 36:873-84
Gehad, Ahmed E; Lichtman, Michael K; Schmults, Chrysalyne D et al. (2012) Nitric oxide-producing myeloid-derived suppressor cells inhibit vascular E-selectin expression in human squamous cell carcinomas. J Invest Dermatol 132:2642-51
Trimble, Cornelia L; Clark, Rachael A; Thoburn, Christopher et al. (2010) Human papillomavirus 16-associated cervical intraepithelial neoplasia in humans excludes CD8 T cells from dysplastic epithelium. J Immunol 185:7107-14
Clark, Rachael A (2010) Skin-resident T cells: the ups and downs of on site immunity. J Invest Dermatol 130:362-70
Clark, Rachael A (2009) Regulation gone wrong: a subset of Sezary patients have malignant regulatory T cells. J Invest Dermatol 129:2747-50
Clark, Rachael A; Fuhlbrigge, Robert C (2009) Immunology and skin disease 2009: frontiers in cutaneous immunology. J Invest Dermatol 129:1849-51
Huang, Susan J; Hijnen, Dirkjan; Murphy, George F et al. (2009) Imiquimod enhances IFN-gamma production and effector function of T cells infiltrating human squamous cell carcinomas of the skin. J Invest Dermatol 129:2676-85
Clark, Rachael A; Huang, Susan J; Murphy, George F et al. (2008) Human squamous cell carcinomas evade the immune response by down-regulation of vascular E-selectin and recruitment of regulatory T cells. J Exp Med 205:2221-34
Clark, Rachael A; Kupper, Thomas S (2007) IL-15 and dermal fibroblasts induce proliferation of natural regulatory T cells isolated from human skin. Blood 109:194-202

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