Abstract
Complement receptor lymphocytes (CRL) were isolated from lymph nodes of rabbits immunized 6 days earlier with keyhole limpet hemocyanin. Isolation was accomplished by rosetting with sheep erythrocytes sensitized with antibody and horse complement (EAC) followed by Centrifugation on Ficoll-Hypaque (F-H). The CRL population comprised about 30% of the starting LNC and was highly enriched for surface immunoglobulin (SIg) bearing cells. By the criterion of [ 3H]thymidine incorporation, CRL responded poorly to Concanavalin A (ConA), but extensively to goat anti-rabbit Fab′ (AFab′) ; indeed, the CRL responded better than unfractionated LNC to AFab′. Upon addition of KLH, the CRL produced much less antibody and total Ig than unfractionated LNC. KLH-challenged T cells prepared on anti-Fab′ columns also synthesized minimal amounts of antibody and Ig. Antigenic challenge of a mixture of CRL and T cells synthesized 3 to 5 times more antibody and Ig than the sum of the individual CRL- and T cell-mediated syntheses. Moreover, the KLH concentration dependency, kinetics, and ratio of 19S 7S antibody responses were similar for the unfractionated LNC and the reconstituted mixture of CRL plus T cells. Thus in all examined parameters the reconstituted responses were indistinguishable from the responses of the unfractionated LNC. Therefore, the CRL and T cell populations can be utilized individually and in combination to study their roles in the in vitro induction and regulation of the anamnestic response to KLH.
Published Version
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