Abstract

This work covers several areas of study of the structure of the murine Ia antigens, which are integral membrane glycoproteins encoded by genes in the I-region of the H-2 complex (MHC) of the mouse. The first objective is the elucidation of the structures of the oligosaccharides carried by Ia antigens. We will complete our studies of the carbohydrate moieties of the alpha and beta chains of the I-A?k? antigen using radiochemical techniques. The second objective is the localization of the antigenic determinants of the I-A?k? antigen. Monoclonal antibodies will be used to define the relationship between the antigenic structure and the primary structure of the I-A?k? antigen, since a large number of reagents are available for this antigen. If possible, this approach will be extended to the I-A?d? antigen. The third objective is to carry out a detailed examination of the orientation of the Ia antigens in the lipid bilayer of the plasma membrane, including the development of markers for the extracellular, cytoplasmic, and membrane-integrating portions of the molecules. Within this general objective are experiments designed to search for potential interactions of Ia antigens with other membrane proteins or with cytoskeletal elements. A corollary of these experiments will be a search for potential interactions between Ia molecules, the foreign antigen, and the antigen-specific receptor of T cells. Experiments to accomplish the three objectives fall within the areas of immunochemistry and biochemistry. Ia antigens will be purified by affinity chromatography on monoclonal antibody columns, chains will be separated by SDS-PAGE or by ion exchange chromatography in acid-urea, and antigenic fragments will be monitored by indirect immunoprecipitation or by RIA. The primary approach to be employed in documenting the interaction of Ia antigens with other molecules will be the use of chemical cross-linkers. (CS)

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA036700-05
Application #
3174274
Study Section
Allergy and Immunology Study Section (ALY)
Project Start
1983-07-01
Project End
1988-06-30
Budget Start
1987-07-01
Budget End
1988-06-30
Support Year
5
Fiscal Year
1987
Total Cost
Indirect Cost

  Institution

Name
National Jewish Health
Department
Type
DUNS #
City
Denver
State
CO
Country
United States
Zip Code
80206

  Publications

Wade, W F; Ward, E D; Rosloniec, E F et al. (1994) Truncation of the A alpha chain of MHC class II molecules results in inefficient antigen presentation to antigen-specific T cells. Int Immunol 6:1457-65
Rosloniec, E F; Beard, K S; Freed, J H (1993) Functional analysis of the antigen binding region of an MHC class II molecule. Mol Immunol 30:491-501
Freed, J H; Marrack, P (1993) Tissue-specific expression of self peptides bound by major histocompatibility complex class II molecules. Chem Immunol 57:88-112
Newell, M K; VanderWall, J; Beard, K S et al. (1993) Ligation of major histocompatibility complex class II molecules mediates apoptotic cell death in resting B lymphocytes. Proc Natl Acad Sci U S A 90:10459-63
Rosloniec, E F; Vitez, L J; Buus, S et al. (1990) MHC class II-derived peptides can bind to class II molecules, including self molecules, and prevent antigen presentation. J Exp Med 171:1419-30
Wade, W F; Freed, J H; Edidin, M (1989) Translational diffusion of class II major histocompatibility complex molecules is constrained by their cytoplasmic domains. J Cell Biol 109:3325-31
Rosloniec, E F; Vitez, L J; Beck, B N et al. (1989) I-Ak polymorphisms define a functionally dominant region for the presentation of hen egg lysozyme peptides. J Immunol 143:50-8
Wade, W F; Chen, Z Z; Maki, R et al. (1989) Altered I-A protein-mediated transmembrane signaling in B cells that express truncated I-Ak protein. Proc Natl Acad Sci U S A 86:6297-301
Rosloniec, E F; Gay, D; Freed, J H (1989) Epitopic analysis by anti-I-Ak monoclonal antibodies of I-Ak-restricted presentation of lysozyme peptides. J Immunol 142:4176-83
Newell, M K; Justement, L B; Miles, C R et al. (1988) Biochemical characterization of proteins that co-purify with class II antigens of the murine MHC. J Immunol 140:1930-8

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