The main goal of this proposal is to purify the Mr 40K colonic epithelial protein to homogeneity, characterize it biochemically, investigate its polymorphism in different tissue and examine its biological role by establishment of an experimental autoimmune model for ulcerative colitis (UC) in mice. Recent data strongly suggest that this protein is a target self-antigen that triggers an autoimmune reaction which may be important in the pathogenesis of UC. The protein is also present in experimental animals, immunogenic in mice and preliminary studies demonstrated gross and histological changes in colonic mucosa upon injection in the appropriate adjuvant. We will characterize the T cell response to the purified Mr 40K protein and will analyze the T cell proliferative responses to different peptides derived from the Mr 40K protein by chemical or enzymatic cleavage followed by purification on HPLC. We will then determine which of the immunogenic peptide(s) induces an immune response leading to inflammation in the colon. The MHC restriction and T cell receptor usage by these T cell clones will be analyzed by two general approaches: a) The Mr 40K protein/peptide(s) will be injected with various adjuvants into susceptible mouse strains (high responder strains) using various protocols developed for the induction of other autoimmune diseases. b) T cell lines/clones will be adoptively transferred into normal and into immune-compromised mice including SCID mice. Disease induction will be assessed by gross examination and histology. Two main questions concerning the immune mechanisms responsible for the onset of the experimental disease will be addressed: a) What is the phenotype of the antigen presenting cell (APC) that presents the Mr 40K protein to the antigen-specific T cell that mediates tissue damage? Irradiation bone marrow chimeras will be constructed such that the hematopoietic compartment is of a different haplotype than the rest of the host's tissues. T cell clones histocompatible with either the hematopoietic or the host tissue will be adoptively transferred into these chimeras to determine whether the APC is a classical bone marrow-derived cell or a colon epithelial cell. b) How is tissue damage inflicted upon the colon epithelium? Three possible mechanisms will be tested: antibody-dependent cell-mediated cytotoxicity.

National Institute of Health (NIH)
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
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General Medicine A Subcommittee 2 (GMA)
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University of Medicine & Dentistry of NJ
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Griffel, L H; Amenta, P S; Das, K M (2000) Use of a novel monoclonal antibody in diagnosis of Barrett's esophagus. Dig Dis Sci 45:40-8
Biancone, L; Mandal, A; Yang, H et al. (1995) Production of immunoglobulin G and G1 antibodies to cytoskeletal protein by lamina propria cells in ulcerative colitis. Gastroenterology 109:3-12
Hassan, T; Kanisawa, Y; Meyers, S et al. (1995) Expression of a unique protein on colon cancer cells that reacts with a novel monoclonal antibody and ulcerative colitis serum. Clin Exp Immunol 100:457-62
Shah, U; Dasgupta, A; Das, K M (1995) Development of ulcerative colitis-associated anti-idiotype antibody using a novel monoclonal antibody against a colonic autoantigen. Cell Immunol 166:154-7
Das, K M; Prasad, I; Garla, S et al. (1994) Detection of a shared colon epithelial epitope on Barrett epithelium by a novel monoclonal antibody. Ann Intern Med 120:753-6
Bhagat, S; Das, K M (1994) A shared and unique peptide in the human colon, eye, and joint detected by a monoclonal antibody. Gastroenterology 107:103-8
Mandal, A; Dasgupta, A; Jeffers, L et al. (1994) Autoantibodies in sclerosing cholangitis against a shared peptide in biliary and colon epithelium. Gastroenterology 106:185-92
Dasgupta, A; Mandal, A; Das, K M (1994) Circulating immunoglobulin G1 antibody in patients with ulcerative colitis against the colonic epithelial protein detected by a novel monoclonal antibody. Gut 35:1712-7
Halstensen, T S; Das, K M; Brandtzaeg, P (1993) Epithelial deposits of immunoglobulin G1 and activated complement colocalise with the M(r) 40 kD putative autoantigen in ulcerative colitis. Gut 34:650-7
Biancone, L; Das, K M; Roberts, A I et al. (1993) Ulcerative colitis serum recognizes the M(r) 40K protein on colonic adenocarcinoma cells for antibody-dependent cellular cytotoxicity. Digestion 54:237-42

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