ELDKWA-epitope-specific monoclonal antibodies inhibit HIV env-mediated syncytium formation

Recent studies demonstrated that the N- and C-domains of HIV-1 gp41 are involved in virus-mediated membrane fusion resulting in HIV-entry into target cells. Synthetic N- and C-domain peptides, such as DP107 and DP178, potently inhibit membrane fusion induced by both laboratory-adapted strains and primary isolates of HIV-1. Monoclonal antibody (mAb) 2F5 recognizing ELDKWA-epitope on the gp41 C-domain has shown broad neutralizing activity to many HIV stains even primary isolates. To test the neutralizing mechanism of the ELDKWA-epitope-specific antibody, mAbs with predefined ELDKWA-epitope-specificity were induced by synthetic epitope-peptide instead of a natural or recombinant gp41 bearing this epitope. All of five mAbs were identified to recognize ELDKWA-epitope on recombinant soluble gp41 in ELISA-assay. In flow cytometry analysis, four out of five mAbs could bind to HIV-Env+ CHO-WT cells and such binding could be inhibited by ELDKWA-epitope peptide, which suggests that these mAbs could recognize native envelope protein expressed on cell membranes. Interestingly, two out of five mAbs could inhibit membrane fusion between the CXCR4- and CD4-expressing 3T3.T4.CXCR4 cells and HIV-Env+ CHO-WT cells in a dose-dependent manner, consistent with their potent binding capability to HIV-Env+ CHO-WT cells. Parren et al. (1998) suggested that neutralization of HIV-1 by antibody may be determined primarily by occupancy of sites and spatial obstruction which blocks virus entry. Based on this theory, a model was suggested to explain the neutralizing activities of mAbs against the epitope ELDKWA.