Under the influence of the thymic environment, which is comprised of bone marrow derived cells and epithelium, several important aspects of T cell differentiation occur, including antigen receptor expression, establishment of MHC-restricted antigen recognition, and self-tolerance. Understanding the contributions of thymic epithelium (TE) to T cell differentiation has been hampered by a lack of information regarding thymic epithelium. The proposed work will generate TE cell lines and use these cell lines to characterize TE biochemically and functionally. Cell lines representative of cortical and medullary TE will be examined for their ability to physically associate with thymocyte subpopulations in vitro, to secrete cytokines affecting thymocyte proliferation/ differentiation, and to present antigen in the context of major histocompatibility complex molecules.These cell lines will be used to generate a panel of monoclonal antibodies specific for cell surface molecules expressed by cortical and medullary TE populations. Anti-TE antibodies will be used to immunochemically characterize the cell surface molecules expressed by different populations of thymic epithelium cells and to identify molecules unique to cortical and medullary thymic epithelial cells. Immunochemical studies of these molecules will be complemented by immunoelectron microscopic studies to precisely define their in situ distribution within the thymus. Finally, by testing the effects of these anti-epithelial antibodies on thymic function in vivo and vitro, the participation of these epithelial cell surface molecules in the process of T cell differentiation will be assessed. These studies will utilize multi-parameter flow cytometry to analyze thymocyte populations differentiating in the presence of anti-TE monoclonal antibodies specific for cortical or medullary TE. The significance of the proposed work is that it will define the thymic environment in terms of the epithelial cell surface and/or cytokines that collectively constitute the thymic environment. Characterization of the constituent components of the thymic environment will lead to a clearer understanding of the process of T cell differentiation and may lead to the design of therapeutic approaches to enhance thymic function.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
2R01AI024137-04A1
Application #
3136830
Study Section
Immunobiology Study Section (IMB)
Project Start
1986-12-01
Project End
1993-06-30
Budget Start
1990-07-01
Budget End
1991-06-30
Support Year
4
Fiscal Year
1990
Total Cost
Indirect Cost
Name
University of Washington
Department
Type
Schools of Medicine
DUNS #
135646524
City
Seattle
State
WA
Country
United States
Zip Code
98195
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Dooley, James; Erickson, Matthew; Larochelle, William J et al. (2007) FGFR2IIIb signaling regulates thymic epithelial differentiation. Dev Dyn 236:3459-71
Gillard, Geoffrey O; Dooley, James; Erickson, Matthew et al. (2007) Aire-dependent alterations in medullary thymic epithelium indicate a role for Aire in thymic epithelial differentiation. J Immunol 178:3007-15
Dooley, James; Erickson, Matthew; Gillard, Geoffrey O et al. (2006) Cervical thymus in the mouse. J Immunol 176:6484-90
Gillard, Geoffrey O; Farr, Andrew G (2006) Features of medullary thymic epithelium implicate postnatal development in maintaining epithelial heterogeneity and tissue-restricted antigen expression. J Immunol 176:5815-24
Fontenot, Jason D; Dooley, James L; Farr, Andrew G et al. (2005) Developmental regulation of Foxp3 expression during ontogeny. J Exp Med 202:901-6
Rodriguez-Galan, Maria Cecilia; Bream, Jay H; Farr, Andrew et al. (2005) Synergistic effect of IL-2, IL-12, and IL-18 on thymocyte apoptosis and Th1/Th2 cytokine expression. J Immunol 174:2796-804
Dooley, James; Erickson, Matt; Farr, Andrew G (2005) An organized medullary epithelial structure in the normal thymus expresses molecules of respiratory epithelium and resembles the epithelial thymic rudiment of nude mice. J Immunol 175:4331-7
Dooley, James; Erickson, Matthew; Roelink, Henk et al. (2005) Nude thymic rudiment lacking functional foxn1 resembles respiratory epithelium. Dev Dyn 233:1605-12
Cooper, Cristine J; Turk, Gail L; Sun, Mingyi et al. (2004) Cutting edge: TCR revision occurs in germinal centers. J Immunol 173:6532-6

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