Characterization of the bovine secondary in vitro antibody response

In the present report, the bovine secondary in vitro antibody response to keyhole limpet hemocyanin is described. The induction of anti-KLH antibody was not dependent upon the presence of mitogen but was antigen specific (KLH vs ovalbumin). Furthermore, this response was dependent upon cell density (10(6) per well), antigen dose (1 to 10(-5) ug/culture) and time in culture (5 days). The antibody produced was specific for KLH as measured in several binding assays. An unresponsive state was detected with high concentrations of KLH (more than 10 ug per culture) which was not due to the formation of immune complexes but to the inactivation of B and/or T cells. The induction of the antibody response was dependent on the presence of macrophages (syngeneic or allogeneic) and their presence could not be replaced with 2-mercaptoethanol.