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Broad and potent neutralization of HIV-1 by a gp41-specific human antibody

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Abstract

Characterization of human monoclonal antibodies is providing considerable insight into mechanisms of broad HIV-1 neutralization. Here we report an HIV-1 gp41 membrane-proximal external region (MPER)-specific antibody, named 10E8, which neutralizes 98% of tested viruses. An analysis of sera from 78 healthy HIV-1-infected donors demonstrated that 27% contained MPER-specific antibodies and 8% contained 10E8-like specificities. In contrast to other neutralizing MPER antibodies, 10E8 did not bind phospholipids, was not autoreactive, and bound cell-surface envelope. The structure of 10E8 in complex with the complete MPER revealed a site of vulnerability comprising a narrow stretch of highly conserved gp41-hydrophobic residues and a critical arginine or lysine just before the transmembrane region. Analysis of resistant HIV-1 variants confirmed the importance of these residues for neutralization. The highly conserved MPER is a target of potent, non-self-reactive neutralizing antibodies, suggesting that HIV-1 vaccines should aim to induce antibodies to this region of HIV-1 envelope glycoprotein.

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Figure 1: Analyses of 10E8 sequence and neutralization.
Figure 2: Binding specificity of 10E8.
Figure 3: Analysis of 10E8 autoreactivity.
Figure 4: Crystal structure of 10E8 Fab in complex with its gp41 MPER epitope.
Figure 5: A site of gp41 vulnerability.

Accession codes

Primary accessions

GenBank/EMBL/DDBJ

Protein Data Bank

Data deposits

The nucleotide sequence of 10E8 heavy and light chains have been submitted to GenBank under accession numbers JX645769 and JX645770. Coordinates and structure factors for 10E8 Fab in complex with the gp41 MPER have been deposited with the Protein Data Bank under accession code 4G6F.

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Acknowledgements

We thank C. W. Hallahan for statistical analyses. We thank K. Lloyd. R. Parks, J. Eudailey and J. Blinn for performing autoantibody assays. We also thank M. Zwick for providing us with the HIV-1 JR2 MPER alanine mutant pseudovirus plasmids. HIV-2/HIV-1 chimaeras were provided by G. Shaw and L. Morris. We thank J. Stuckey for assistance with figures, and members of the Structural Biology Section and Structural Bioinformatics Core, Vaccine Research Center, for discussions and comments on the manuscript. This project has been funded in part with federal funds from the Intramural Research Programs of NIAID and the National Cancer Institute, National Institutes of Health, under Contract no. HHSN261200800001E. Use of sector 22 (Southeast Region Collaborative Access team) at the Advanced Photon Source was supported by the US Department of Energy, Basic Energy Sciences, Office of Science, under contract number W-31-109-Eng-38. The content of this publication does not necessarily reflect the views or policies of the Department of Health and Human Services, nor does mention of trade names, commercial products, or organizations imply endorsement by the US Government.

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M.C., J.H., L.L., G.O., J.R.M. and P.D.K. designed the study, analysed the data, and prepared this manuscript. J.H. and L.L. performed B-cell sorting, antibody cloning, epitope mapping assay, MPER-specific neutralizing sera screening and assessed the impact of sequence variation on 10E8 neutralization. M.K.L. and J.R.M. tested the breadth and potency of 10E8. B.C., S.K.S. and R.W. performed the infected cell surface staining and antibody-virion washout assays. S.M.A. and B.F.H. performed the autoreactivity assays. G.O., Y.Y. and P.D.K. performed 10E8 structural analysis, with T.W. and B.Z. assisting with paratope alanine scanning. R.T.B. screened the B-cell culture supernatants for neutralization activity. H.I. sequenced the patient N152 virus. S.A.M. led the clinical care of the patients. M.C., L.L., N.A.D.-R. and N.S.L. optimized B-cell culture protocol.

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Correspondence to Mark Connors.

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Huang, J., Ofek, G., Laub, L. et al. Broad and potent neutralization of HIV-1 by a gp41-specific human antibody. Nature 491, 406–412 (2012). https://doi.org/10.1038/nature11544

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