Primary in vitro immunization with multimeric synthetic peptides of HIV-1 envelope glycoproteins: generation of neutralizing human monoclonal antibodies.

Peripheral blood lymphocytes from healthy HIV-1 seronegative donors were immunized in vitro with the following synthetic peptides: (i) an octameric poly-L-lysine conjugated peptide of the HIV-1MN V3 loop and (ii) a resin bound synthetic peptide aa642-665 of HIV-1 gp41. Lymphoblastoid cell lines (LCL) were obtained by immortalization with Epstein-Barr virus (EBV). We produced four LCL secreting human monoclonal antibodies (HuMoAbs) of the IgM isotype: three were directed against the V3 domain (FC10, FC81 and CF41) and one against aa642-665 (CA45C). Two of these HuMoAbs (FC81 and CA45C) reacted to viral surface antigen on HIV-1-infected cells. All the HuMoAbs inhibited 40-53% of cell fusion induced by HIV-1-infected H9 cells at 5 micrograms/ml. They also neutralized, at lower concentrations, cell-free infection with HIV-1MN, HIV-1IIIB and four primary clinical HIV-1 isolates. No enhancing activity of the HuMoAbs in the presence of complement was observed. The results presented here show the feasibility of generating neutralizing human monoclonal antibodies against HIV-1 by primary in vitro immunization with selected synthetic peptides of HIV-1 envelope glycoproteins. This approach has provided tools for further studies of synergistic neutralization assays, and generated potential immunoglobulin candidates for passive immunotherapy.