Our objective is to develop a rational approach to specific desensitization of HLA class I-sensitized patients by means of anti-idiotypic antibodies (Ab-2). The project is divided into two parts: I) characterization of the memory B cell response to HLA-A2 and II) development of therapeutic anti- idiotypic reagents. The first section, comprising the bulk of the work to be done, consists of 4 inter-related components: 1) production of mg quantities of soluble HLA class I antigens by transfer into a class I """"""""null"""""""" mutant human lymphoblastoid cell of HLA-A2 alpha chain genes encoding a culture supernatant; 2) coupling of purified HLA-A2 to solid supports for affinity purification of A2-specific IgG antibodies and B cells from A2-sensitized patients; 3) analysis using HLA-A2 subtype gene transferent LCL, mouse monoclonal antibodies to HLA-A2 antigens, and polyclonal anti-idiotypic antibodies (Ab-2) of the epitope and idiotope specificity of the antibody response to HLA-A2; and 4) in vitro monitoring of anti-HLA-A2 memory B cell responses in transfused patients, and leukapheresis of selected A2-sensitized patients to provide sufficient B cells for in vitro testing of Ab-2 preparations. The information generated by part I of the project will be used to develop therapeutic Ab-2 preparations. The goal of part II will be to generate mouse monoclonal antibodies with specificity for """"""""public"""""""" or cross-reactive idiotopes of A2-specific memory B cells. These Ab-2 will be tested as is, or coupled to ricin-A chain and tested for efficacy as immunotoxins using EBV-transformed, HLA-A2-specific memory B cell lines, and bulk cultures of HLA-A2-specific B cells and T helper cells for sensitized patients.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI026941-03
Application #
3141033
Study Section
Surgery, Anesthesiology and Trauma Study Section (SAT)
Project Start
1989-04-01
Project End
1994-03-31
Budget Start
1991-04-01
Budget End
1992-03-31
Support Year
3
Fiscal Year
1991
Total Cost
Indirect Cost
Name
University of Wisconsin Madison
Department
Type
Schools of Medicine
DUNS #
161202122
City
Madison
State
WI
Country
United States
Zip Code
53715
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Burlingham, W J; Jankowska-Gan, E; DeVito-Haynes, L et al. (1998) HLA (A*0201) mimicry by anti-idiotypic monoclonal antibodies. J Immunol 161:6705-14
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Niguma, T; DeVito, L D; Grailer, A P et al. (1993) HLA-A2-specific antibody production in severe combined immunodeficient mice reconstituted with human peripheral blood leukocytes from HLA-presensitized donors. Transplant Proc 25:239-40
De Vito, L D; Mason, B P; Jankowska-Gan, E et al. (1993) Epitope fine specificity of human anti-HLA-A2 antibodies. Identification of four epitopes including a haptenlike epitope on HLA-A2 at lysine 127. Hum Immunol 37:165-77

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