A fusion intermediate gp41 immunogen elicits neutralizing antibodies to HIV-1 [Protein Structure and Folding]

August 26th, 2014 by Lai, R. P. J., Hock, M., Radzimanowski, J., Tonks, P., Lutje Hulsik, D., Effantin, G., Seilly, D. J., Dreja, H., Kliche, A., Wagner, R., Barnett, S. W., Tumba, N., Morris, L., LaBranche, C. C., Montefiori, D. C., Seaman, M. S., Heeney, J. L., Weissen

The membrane proximal external region (MPER) of the HIV-1 Env glycoprotein subunit gp41 is targeted by potent broadly neutralizing antibodies (bnAb) 2F5, 4E10 and 10E8. These antibodies recognize linear epitopes and have been suggested to target the fusion intermediate conformation of gp41 that bridges viral and cellular membranes. Anti-MPER antibodies exert different degrees of membrane interaction, which is considered to be the limiting factor for the generation of such antibodies by immunization. Here we characterize a fusion intermediate conformation of gp41 (gp41int-cys) and show that it folds into an elongated ~12 nm long extended structure based on small angle X-ray scattering data. Gp41int-cys was covalently linked to liposomes via its C-terminal cysteine and employed as immunogen. The gp41int-cys proteoliposomes were administered alone or in prime-boost regimen with trimeric Env gp140CA018 in guinea pigs and elicited high anti-gp41 IgG titers. The sera interacted with a peptide spanning the MPER region, demonstrated competition with bnAbs 2F5 and 4E10 and exerted modest lipid binding, indicating the presence of MPER-specific antibodies. Although the neutralization potency generated solely by gp140CA018 was higher than that induced by gp41int-cys, the majority of animals immunized with gp41int-cys proteoliposomes induced modest breadth and potency in neutralizing tier 1 pseudoviruses and replication-competent SHIVs in the TZM-bl assay as well as responses against tier 2 HIV-1 in the A3R5 neutralization assay. Our data thus demonstrate that liposomal gp41 MPER formulation can induce neutralization activity and the strategy serves to improve breadth and potency of such antibodies by improved vaccination protocols.