Relative frequencies of secondary B cells activated by cognate vs. other mechanisms

Three distinct mechanisms for the activation of secondary B cells to antibody-forming cells have been examined in splenic fragment cultures. The clonal response to a protein antigen [cytochrome c (cyt)] was quantified in terms of both the number of B cells activated and the amount of antibody produced by each clone. The vast majority of memory B cells required surface immunoglobulin (sIg) receptor-mediated uptake of antigen followed by cognate interactions with an antigen-specific T helper cell. Nevertheless, a significant number (as many as 12%) could be activated via soluble factor-mediated bystander activation, involving occupation of neither the sIg receptor nor class II major histocompatibility complex (MHC) molecules. This pathway, however, was relatively inefficient in that individual clones secreted less than half as much antibody as clones activated as a result of cognate collaboration with a T helper cell. A similar number of secondary B cells were activated following nonspecific uptake of high concentrations of antigen for which splenic fragment T cells had been primed [i.e., hemocyanin (Hy)], independent of sIg receptor occupancy. Antibody levels were similar to those in cultures where B cells were activated in a cognate manner with sIg receptor occupancy. When Hy-stimulated fragment cultures were supplemented with polymerized cyt, the frequency of activation via this latter pathway increased fourfold, despite the fact that no cyt-primed T cells were present. This observation supports the idea that receptor-mediated uptake of antigen serves not just to focus antigen but also provides an important signal in activating B cells. Bystander B cell activation, however, was not enhanced by providing a sIg cross-linking signal with polymerized cyt. The lower level of antibody production by B cells activated in a bystander fashion and the inability to enhance their frequency of activation with sIg receptor occupancy suggest that there is indeed a fundamental difference between soluble factor-mediated bystander activation and activation via T cell determinant-mediated cognate T helper-B cell interactions, perhaps involving signaling through class II MHC molecules.