The overall objective of this proposal is to define the immunoregulatory role of anti-idiotypic antibodies to anti-HLA. Using a microcytotoxicity inhibition assay, we have recently obtained evidence for development of anti-HLA antibodies followed by anti-idiotypic antibodies to HLA class 1 and 11 antibodies in human liver allograft recipients. The proposed research will define the fine specificities of anti-anti-HLA activity using multiple monospecific anti-HLA reagents in order to determine the extent of idiotypic cross-reactivity that exists between antibodies that define distinct polymorphisms of MHC class 1 and class 11 antigens. In particular, these studies will define the extent to which antibodies directed to cross-reactive HLA epitopes (CREGs) express cross-reactive idiotopes. Our preliminary studies Indicate that these anti-idiotypic antibodies to anti-HLA antibodies bind to autologous stimulated T cell and possibly to the clonotypic T cell structures. We will determine the ability of pre- and post- transplant sera to bind to autologous normal and activated T lymphocytes, including biopsy derived T cell clones, using immunofluorescence and flow cytometry. We will use selected sera which bind to T cells in immunoprecipitation analysis, to define the immunochemical nature of the membrane molecules reactive to the anti-idiotypic antibodies. Since anti-idiotypic antibodies to anti-HLA antibodies are reactive with T cells possibly via T cell clonotypic structures, we will determine the immunoregulatory effects of these antibodies on specific cellular immune functions. We will analyze the capacity of anti-idiotypic antibodies to block mixed lymphocyte culture responses of recipients against donor antigens, to block cytotoxic T lymphocyte generation and killing, and to Inhibit proliferation and/or killing by biopsy derived cloned T lymphocytes. Finally, we will develop methods with which to establish anti-anti-HLA secreting B cells; these will include transformation of B cells secreting antibodies using Epstein-Barr virus and/or fusion with mouse myeloma partners. The long term goal of this study is to further define the role of anti-idiotypic antibodies in the regulation of human immune responses so that idiotypic manipulation of immune response to transplantation and tumor antigens can become a viable goal.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI028356-05
Application #
3142824
Study Section
Surgery, Anesthesiology and Trauma Study Section (SAT)
Project Start
1989-07-01
Project End
1994-06-30
Budget Start
1993-07-01
Budget End
1994-06-30
Support Year
5
Fiscal Year
1993
Total Cost
Indirect Cost
Name
Washington University
Department
Type
Schools of Medicine
DUNS #
062761671
City
Saint Louis
State
MO
Country
United States
Zip Code
63130
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Chauhan, B; Phelan, D L; Marsh, J W et al. (1993) Characterization of antiidiotypic antibodies to donor HLA that develop after liver transplantation. Transplantation 56:443-8
Ratner, L E; Phelan, D; Brunt, E M et al. (1993) Probable antibody-mediated failure of two sequential ABO-compatible hepatic allografts in a single recipient. Transplantation 55:814-9
Mathew, J M; Marsh, J W; Susskind, B et al. (1993) Analysis of T cell responses in liver allograft recipients. Evidence for deletion of donor-specific cytotoxic T cells in the peripheral circulation. J Clin Invest 91:900-6
Duffy, B F; Mathew, J M; Flye, M W et al. (1993) Development of autoantibodies to T cell clonotypic structures in a liver-kidney allograft recipient. Transplantation 56:212-6
Phelan, D; Hadley, G; Duffy, B et al. (1991) Antiidiotypic antibodies to HLA class I alloantibodies in normal individuals: a mechanism of tolerance to noninherited maternal HLA antigens. Hum Immunol 31:1-6
Hadley, G A; Phelan, D; Duffy, B F et al. (1990) Lack of T-cell tolerance of noninherited maternal HLA antigens in normal humans. Hum Immunol 28:373-81