Primary biliary cirrhosis (PBC) and autoimmune hepatitus (AI-CAH) ar two chronic diseases of the liver, believed to be autoimmune in nature, with etiologies and pathogenesis that remain unknown. These disorders affect predominantly women, mostly in th prime of their life. For example, over 90% of patients with PBC are women between the ages of 20-70. Liver transplantation ha s proven to be an effective treatment for both AI-CAH and PBC. However, disease recurrence remains an important concern. The goal of this project is to elucidate the role of cytokines and cytotoxic mediators in autoimmune liver disease. Specifically, we propose to: 1) characterize the cytokine profiles within the liver of patients with PBC and AI-CAH 20 identify the cytotoxic mediators 30 characterize the functional properties of lymphoid cells that have infiltrated the liver and 40 apply this information to establish immunological parameters that distinguish disease recurrence from allograft rejection. Molecular methodologies will be utilized to determine and compare the cytokine arrays in normal liver and liver explant tissue from patients with PBC and AI-CAH. RNA obtained from liver tissue will be reverse transcribed, and analyzed for cytokine gene expression using semi- quantitative PCR. In situ hybridization and immunohistochemical techniques will identify the cellular source of cytokine message within the liver. Evidence for specific cellular pathways of liver tissue injury will be obtained by immunohistochemical techniques and Western blot hybridization analysis for serine esterases, perforin, eosinophil cytotoxic proteins and autoantibodies. Direct in vitro examination of infiltrating cells recovered from the PBC and AI-CAH liver will establish the diversity of T cells within the infiltrate, the cytotoxic activity and specificity of T lymphocytes, the frequency of specific cytokine producing cells, the arrays of intrahepatic autoantibodies, and the presence of non-specific effector cells including NK cells and eosinophils. The effect of recombinant cytokines on NHC antigen expression by target tissue will be tested on epithelial and hepatocyte- derived cell lines. Finally, patterns of cytokine production in PBC and AI-CAH will be compared with those of patients who have received liver transplants because of autoimmune liver disease in an effort to distinguish disease recurrence from allograft rejection. Taken together, these studies will provide important mechanistic insights into these two enigmatic autoimmune diseases as well as clarify th issue of disease recurrence post-orthotopic transplantation.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
First Independent Research Support & Transition (FIRST) Awards (R29)
Project #
5R29AI035994-05
Application #
2517252
Study Section
Immunological Sciences Study Section (IMS)
Project Start
1993-09-01
Project End
1999-08-31
Budget Start
1997-09-01
Budget End
1999-08-31
Support Year
5
Fiscal Year
1997
Total Cost
Indirect Cost
Name
Stanford University
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
800771545
City
Stanford
State
CA
Country
United States
Zip Code
94305
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Krams, S M; Martinez, O M (1998) Apoptosis as a mechanism of tissue injury in liver allograft rejection. Semin Liver Dis 18:153-67
Krams, S M; Fox, C K; Beatty, P R et al. (1998) Human hepatocytes produce an isoform of FAS that inhibits apoptosis. Transplantation 65:713-21
Hayashi, M; Keeffe, E B; Krams, S M et al. (1998) Allograft rejection after liver transplantation for autoimmune liver diseases. Liver Transpl Surg 4:208-14
Krams, S M; Cao, S; Hayashi, M et al. (1996) Elevations in IFN-gamma, IL-5, and IL-10 in patients with the autoimmune disease primary biliary cirrhosis: association with autoantibodies and soluble CD30. Clin Immunol Immunopathol 80:311-20
Martinez, O M; Villanueva, J C; Gershwin, M E et al. (1995) Cytokine patterns and cytotoxic mediators in primary biliary cirrhosis. Hepatology 21:113-9
Hayashi, M; Martinez, O M; Garcia-Kennedy, R et al. (1995) Expression of cytokines and immune mediators during chronic liver allograft rejection. Transplantation 60:1533-8
Krams, S M; Egawa, H; Quinn, M B et al. (1995) Apoptosis as a mechanism of cell death in liver allograft rejection. Transplantation 59:621-5
Krams, S M; Villanueva, J C; Quinn, M B et al. (1995) Expression of the cytotoxic T cell mediator granzyme B during liver allograft rejection. Transpl Immunol 3:162-6