IN-VITRO IMMUNIZATION OF MOUSE SPLEEN-CELLS FOR THE PRODUCTION OF MONOCLONAL IGG1 ANTIBODIES USING AN ANTIGEN-SPECIFIC T-HELPER CELL CLONE (D.10.G4.1)

The in vitro immunization of mouse spleen cells for the production of antigen-specific monoclonol antibodies has been investigated in detail . Using various, previous published in vitro immunization protocols wi th and without antigen, we were able to show that IgM antibody product ion was not induced by the antigen. Nevertheless we developed a new in vitro immunization protocol for eliciting antigen-specific monoclonal IgG1 antibodies. Specific antibodies were generated by an antigen-dri ven immune response which included an immunoglobulin class switch. Co- culture of mouse spleen cells with an activated conalbumin specific T helper cell clone (T(H)2 D.10.G4.1) in the presence of conalbumin elic ited three monoclonal IgG1 antibodies that bound specifically to conal bumin. We conclude that for the in vitro generation of monoclonal anti bodies against T cell dependent proteins the addition of antigen activ ated specific T(H)2 cell clones is essential.