Abstract
Normal BALB/c spleen cells are unresponsive in vitro to the phosphorylcholine (PC) determinant in the presence of anti-idiotype antibodies specific for the TEPC-15 myeloma protein (T15) which carries an idiotypic determinant indistinguishable from that of most anti-PC antibodies in BALB/c mice. The possibility that idiotype-specific suppressor cells may be generated during the culture period was examined by coculturing the cells with untreated syngeneic spleen cells. Cells that had been preincubated with anti-T15 idiotype (anti-T15id) antibodies and a PC-containing antigen, R36a for 3 d, were capable of specifically suppressing the anti-PC response of fresh normal spleen cells, indicating that idiotype-specific suppressor cells were generated during the culture period. The presence of specific antigen also appeared to be necessary because anti-T15id antibodies and a control antigen, DNP-Lys-Ficoll, were not capable of generating such suppressor cells. Suppressor cells were induced only in the population of spleen cells nonadherent to nylon wool and the suppressive activity was abrogated by treatment with anti-Thy 1.2 serum and complement. These results indicate that anti-idiotype antibodies and specific antigen can generate idiotype-specific suppressor T cells in vitro. These in vitro results may reflect in vivo mechanisms of idiotype suppression.
Highlights
Materials and MethodsMice. 7- to 10-wk-old BALB/e mice were purchased from Cumberland View Farms, Clinton, Tenn
The presence of specific antigen appeared to be necessary because anti-T 15id antibodies and a control antigen, DNP-Lys-FicolI, were not capable of generating such suppressor cells
Suppressor cells were induced only in the population of spleen cells nonadherent to nylon wool and the suppressive activity was abrogated by treatment with anti-Thy 1.2 serum and complement. These results indicate that anti-idiotype antibodies and specific antigen can generate idiotype-specific suppressor T cells in vitro
Summary
Mice. 7- to 10-wk-old BALB/e mice were purchased from Cumberland View Farms, Clinton, Tenn. Anti-Tl5 antibodies were prepared in the ascitic fluid of A/He mice by multiple injections ofT15 protein according to the method of Tung et al [10]. Pretreatment of normal BALB/e spleen cells with the anti-Ig serum (1:80) plus complement resulted in 92% reduction of the PFC to PC after in vitro stimulation. Normal BALB/c spleen cells (1.2 × 107) were cultured for 3 d in a multiwell tissue culture plate (Falcon 3008, Falcon Labware, Div. of Becton, Dickinson & Co., Oxnard, Calif.) with a 1:2,000 final dilution of anti-T15id ascites and 2.5 × 106 R36a cells as described above. Spleen ceils immunized with R36a for 4 d in vitro were incubated for 1 h with a 1:400 final dilution of anti-T15id ascites in BSS. The number of plaques was counted after an additional 1-h incubation with complement, and compared with the number of plaques developed in the absence of anti-T 15id antibody
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