Antigen-specific memory and virgin B cells differ in their requirements for conjugation to T cells

The physical interaction between carrier-specific T hybridoma cells and long-term primed, TNP-specific memory B cells (TNP-MABC) exposed to TNP-OVA was compared to that of unprimed, TNP-specific virgin B cells (TNP-ABC). The direct conjugation of the T and B cells was visualized at the light microscopic level and the number of T/B conjugates was quantified directly. The results demonstrate that the TNP-MABC, as compared to TNP-ABC, formed T/B conjugates after a shorter exposure time to antigen and at a 10-fold lower concentration of antigen. Conjugate formation was inhibited almost completely by treating the TNP-MABC with concentrations of chloroquine that only partially inhibited the ability of the TNP-ABC to form conjugates. Exposure of the T hybridoma cells or the TNP-ABC to monoclonal antibodies directed against cell surface antigens prior to conjugation indicated that L3T4, Thy-1.2, and LFA-1 antigens on the T cells and LFA-1 and I-A antigens on the TNP-ABC are involved in conjugate formation. However, in contrast to the TNP-ABC where treatment of the B cells with anti-LFA-1 blocked T/B conjugate formation, pretreatment of the TNP-MABC with anti-LFA-1 had no effect.