Immune responses in vitro. IV. Suppression of primary M, G, and A plaque-forming cell responses in mouse spleen cell cultures by class-specific antibody to mouse immunoglobulins

Abstract

The suppressive effects of monospecific goat anti-mouse globulins on primary immunoglobulin class-specific plaque-forming cell responses in mouse spleen cell cultures were investigated. Anti-micro suppressed responses in all immunoglobulin classes, whereas anti-gamma(1) and anti-gamma(2) suppressed the gamma(1) and gamma(2) responses but not gammaM or gammaA responses, and anti-gammaA suppressed only gammaA responses. The mechanism of action of the anti-micro was studied in detail because of its suppression of responses in all immunoglobulin classes. The anti-micro was specific for micro-chain determinants; its activity was dose dependent, but was not mediated by killing cells with surface micro-chain determinants. Free gammaM but not gammaG myeloma proteins in solution effectively competed with micro-bearing cells for the anti-micro. An excess of anti-micro was necessary in the cultures for 48 hr to insure complete suppression of 5-day responses. However, after removal of excess anti-micro at 48 hr, responses could be stimulated by newly added antigen in cultures where incubation was prolonged to 7 days. Anti-micro was most effective when added at the initiation of cultures and had no suppressive effect when added at 48 hr. Excess antigen did not effectively compete with anti-micro for antigen receptors. Precursors of antibody-forming cells were shown to be the cell population where the suppressive activity of anti-micro was mediated. The experiments suggest that anti-micro combines with micro-chain determinants in antigen-specific receptors on the surfaces of antibody-forming cell precursors, prevents effective stimulation by antigen and subsequent antibody production. To explain suppression of responses in all Ig classes by anti-micro, several models were proposed. It is not possible to determine from the data whether stimulation of precursor cells with gammaG or gammaA receptors requires concommitant stimulation of separate cells with only gammaM receptors, or whether cells bearing gammaM receptors are precommitted to or differentiate into cells capable of synthesis of other Ig classes, or whether receptors of gammaM and another Ig class are present on some virgin precursors or the second Ig receptor appears after antigenic stimulation.