GENERATION OF HUMAN MONOCLONAL-ANTIBODIES AGAINST HIV-1 PROTEINS - ELECTROFUSION AND EPSTEIN-BARR-VIRUS TRANSFORMATION FOR PERIPHERAL-BLOODLYMPHOCYTE IMMORTALIZATION

Electrofusion and EBV transformation were studied by immortalizing hum an PBLs from blood of HIV-1-positive volunteers. A panel of 33 cell li nes producing human monoclonal antibodies (Hu-MAbs) against HIV-1 was established by cell fusion or EBV transformation. For the first fusion experiments the source of B lymphocytes was peripheral blood of HIV-1 -infected donors in CDC stages II or III with CD4 cell counts higher t han 500/mm(3). Later on, from these patients only, those with high ant i-HIV titers were chosen as blood donors. By that means the yield of s table specific hybridomas was increased twofold. In our experiments el ectrofusion turned out to be a more efficient immortalization method t han EBV transformation, due to a high and constant immortalization rat e. The hybridomas were stable after intensive subcloning and could be cultivated over a period of 8 months without loss in monoclonal antibo dy production. Immunoglobulin class, subtype, reactivity against HIV-1 proteins, Western blot patterns, immunofluorescence, and epitopes wer e characterized. The subtype of all antibodies was IgG(1) or IgG(3). T he light chain was predominantly kappa. All antibodies showed reactivi ty against HIV-1 envelope or core protein. All hybridomas were stable and suited for mass production. Several Hu-MAbs are becoming an import ant tool in the field of diagnosis, research, and immunotherapy.