T cell 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 #
5K08AI060890-03
Application #
7357513
Study Section
Allergy & Clinical Immunology-1 (AITC)
Program Officer
Prograis, Lawrence J
Project Start
2006-02-01
Project End
2010-01-31
Budget Start
2008-02-01
Budget End
2009-01-31
Support Year
3
Fiscal Year
2008
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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