Antigen-specific activation of B cells in vitro after recruitment of T cell help with superantigen

Human B cells can proliferate in vitro after stimulation with anti-Ig and via the CD40 molecule. Superantigens like SEA which bind to MHC class II antigens on, e.g. B cells can polyclonally activate T cells via interaction with their TcR. The activated T cell subsequently activates the B cells to proliferation and Ig-production. To investigate whether superantigen could be used to direct polyclonal T cell help to human B cells stimulated by antigen in a restricted manner resulting in production of antigen-specific antibodies in vitro. Purified B cells were preincubated with the antigen in manners allowing crosslinking of surface-Ig. The antigen exposed B cells were then cultured together with autologous CD4+ helper T cells and in the presence of various concentrations of SEA. Antibody production was measured by ELISA after 7-12 days of culture. Antigen-specific activation of B cells could be obtained after stimulating the B cells with antigen or anti-surface-Ig antibodies in the presence of T helper cells and SEA. The degree of B cell activation (proliferation as well as antibody production) depended on the dose of antigen as well as on the dose of SEA used. Increased crosslinking of surface-Ig on antigen-specific B cells enhanced Ig production. Specific antibody production to a secondary recall antigen (tetanus toxoid) and to primary antigens (DNP and GM2) were obtained. The specific B cell response was dependent on contact between T and B cells. the results obtained demonstrate that the superantigen SEA can recruit T cell help to human B cells specifically stimulated by antigens, resulting in production of antigen reactive antibodies in vitro.