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- Title
Production and characterization of monoclonal antibodies recognizing the α-chain subunits of human Ia alloantigens.
- Authors
Adams, T. E.; Bodmer, Julia G.; Bodmer, W. F.
- Abstract
Two monoclonal antibodies, TAL-l85 and TAL-3C3. specific for human Ia α-chain subunits have been produced by fusing P3/NSJ/l-Ag4-l mouse myeloma cells with spleen cells from a BA LB/c mouse immunized with purified α-chains. Specificity for the α-chain subunits was initially established using a solid-phase radioimmunoassay. Indirect binding assays demonstrated that TAL-1B5 bound strongly to all human B lymphoblastoid lines tested and to CLLs, but only weakly to PBL-B cells and not to PBL-T cells or the T-cell lines Molt 4 and HSB-2. TAL-3C3 bound only weakly to B lymphoblastoid lines and not to CLLs or PBL-B cells. From 125 I cell surface-labelled lysates TAL-1B5 immunoprecipitated a 33,000(α):28,000(β) Ia dimer, but TAL-3C3 failed to immunoprecipitate cell surface molecules. Under denaturing conditions, however. both TAL-lB5 and TAL-3C3 immunoprecipitated the 33,000 2-chain subunit. Competitive inhibition studies demonstrated that both monoclonal antibodies recognize the same or spatially related a-chain antigenic determinants with some slight cross-reactivity against β-chains. 2D-NEPHGE/SDS-PAGE analysis of TAL-l85 immunoprecipitates from [35S)-methionine biosynthetically labelled cells revealed the presence of a number of α-chain spots in association with β-chain products of three previously described loci (β-I, β-2, β-3) suggesting that this antibody recognizes an antigenic site common to those human Ia s-chains so far identified.
- Subjects
IMMUNOGLOBULINS; MONOCLONAL antibodies; LYMPHOCYTES; B cells; CELL membranes; LYMPHOID tissue; SPLEEN; CELL culture; EPITOPES
- Publication
Immunology, 1983, Vol 50, Issue 4, p613
- ISSN
0019-2805
- Publication type
Article