Selective elimination of a B cell subset having acceptor site(s) for T cell-replacing factor (TRF) with biotinylated antibody to the acceptor site(s) and avidin-ricin A-chain conjugate.
A covalent conjugate of avidin with ricin subunit A-chain (avidin-RA) was prepared by using N-succinimidyl 3-(2-pyridyldithio)propionate as a coupling agent. Selective cytotoxic activity after the combined treatment of spleen cells with biotinylated antibody and avidin-RA was demonstrated by the fact that the responsiveness to LPS was selectively abrogated by pretreatment of the cells with biotinylated rabbit anti-mouse immunoglobulin (MIg) antibody, but not with biotinylated anti-Thy-1.2 antibody. Neither the biotinylated antibody alone nor avidin-RA alone was effective in decreasing the responses to mitogens. Moreover, a high anti-DNP PFC response elicited by DNP-KLH-primed BALB/c mouse spleen cells stimulated in vitro with DNP-KLH was mostly abrogated by the pretreatment of the cells with biotinylated anti-MIg antibody and avidin-RA. Again, neither the biotinylated antibody alone nor avidin-RA alone was effective in decreasing the anti-DNP PFC response. This cell-killing method with the use of biotinylated antibody and avidin-RA was applied and evaluated in experimental systems in which the helper action of T cells on B cells was mediated by T cell-replacing factor (TRF) or was performed by the direct interaction of T cells with B cells (cognate interaction). When DNP-KLH-primed splenic B cells, pretreated with biotinylated F(ab')2 fragment of DCF1 male anti-BALB/c-B IgG antibody against acceptor site(s) for TRF followed by treatment with avidin-RA, were stimulated with DNP-OVA in the presence of monoclonal TRF, the anti-DNP PFC response was significantly decreased, whereas the same treated B cells responded well to stimulation with DNP-PPD in the presence of Tbc-primed T cells (cognate interaction). These results indicate that B cells responsible for the cognate interaction and those having TRF acceptor site(s) belong to a distinct subpopulation of B cells, and that the cytocidal action of the noncovalent conjugate of the antibody and RA formed from the biotinylated antibody and avidin-RA via an avidin-biotin complex has immunologic selectivity, eliminating only the latter subset of B cells recognized by the antibody.
D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000502:physiology*; D001360:Avidin / Q000494:pharmacology; D001402:B-Lymphocytes / Q000145:classification / Q000276:immunology / Q000378:metabolism*; D001710:Biotin / Q000378:metabolism; D005260:Female; D007140:Immunoglobulin Fab Fragments / Q000276:immunology; D007165:Immunosuppression Therapy; D015848:Interleukin-5; D008213:Lymphocyte Activation; D008211:Lymphocyte Cooperation; D008212:Lymphocyte Depletion; D008222:Lymphokines / Q000276:immunology / Q000378:metabolism*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008811:Mice, Inbred DBA; D011947:Receptors, Antigen, B-Cell / Q000378:metabolism; D011971:Receptors, Immunologic / Q000276:immunology*; D015375:Receptors, Interleukin-2; D012276:Ricin / Q000494:pharmacology; D013154:Spleen / Q000166:cytology
D000888:Antibodies, Anti-Idiotypic; D007140:Immunoglobulin Fab Fragments; D015848:Interleukin-5; D008222:Lymphokines; D011947:Receptors, Antigen, B-Cell; D011971:Receptors, Immunologic; D015375:Receptors, Interleukin-2; D001360:Avidin; D001710:Biotin; D012276:Ricin