Article | Published:

Distinct conformational states of HIV-1 gp41 are recognized by neutralizing and non-neutralizing antibodies

Nature Structural & Molecular Biology volume 17, pages 14861491 (2010) | Download Citation

Abstract

HIV-1 envelope glycoprotein gp41 undergoes large conformational changes to drive fusion of viral and target cell membranes, adopting at least three distinct conformations during the viral entry process. Neutralizing antibodies against gp41 block HIV-1 infection by targeting gp41′s membrane-proximal external region in a fusion-intermediate state. Here we report biochemical and structural evidence that non-neutralizing antibodies, capable of binding with high affinity to an immunodominant segment adjacent to the neutralizing epitopes in the membrane-proximal region, recognize a gp41 conformation that exists only when membrane fusion is complete. We propose that these non-neutralizing antibodies are induced in HIV-1–infected individuals by gp41 in a triggered, postfusion form and contribute to production of ineffective humoral responses. These results have important implications for gp41-based vaccine design.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Accessions

Primary accessions

Protein Data Bank

Referenced accessions

Protein Data Bank

References

  1. 1.

    Viral membrane fusion. Nat. Struct. Mol. Biol. 15, 690–698 (2008).

  2. 2.

    & The HIV-1 envelope glycoproteins: fusogens, antigens, and immunogens. Science 280, 1884–1888 (1998).

  3. 3.

    et al. Major glycoprotein antigens that induce antibodies in AIDS patients are encoded by HTLV-III. Science 228, 1091–1094 (1985).

  4. 4.

    et al. Characterization of gp41 as the transmembrane protein coded by the HTLV-III/LAV envelope gene. Science 229, 1402–1405 (1985).

  5. 5.

    Mechanism of membrane fusion by viral envelope proteins. Adv. Virus Res. 64, 231–261 (2005).

  6. 6.

    & HIV entry and its inhibition. Cell 93, 681–684 (1998).

  7. 7.

    & Novel therapies based on mechanisms of HIV-1 cell entry. N. Engl. J. Med. 348, 2228–2238 (2003).

  8. 8.

    , , , & A synthetic peptide inhibitor of human immunodeficiency virus replication: correlation between solution structure and viral inhibition. Proc. Natl. Acad. Sci. USA 89, 10537–10541 (1992).

  9. 9.

    et al. A fusion-intermediate state of HIV-1 gp41 targeted by broadly neutralizing antibodies. Proc. Natl. Acad. Sci. USA 105, 3739–3744 (2008).

  10. 10.

    , , & Core structure of gp41 from the HIV envelope glycoprotein. Cell 89, 263–273 (1997).

  11. 11.

    , , , & Atomic structure of the ectodomain from HIV-1 gp41. Nature 387, 426–430 (1997).

  12. 12.

    , , & Rapid evolution of the neutralizing antibody response to HIV type 1 infection. Proc. Natl. Acad. Sci. USA 100, 4144–4149 (2003).

  13. 13.

    et al. HIV-1 evades antibody-mediated neutralization through conformational masking of receptor-binding sites. Nature 420, 678–682 (2002).

  14. 14.

    et al. Antibody neutralization and escape by HIV-1. Nature 422, 307–312 (2003).

  15. 15.

    , , & Neutralizing antibodies generated during natural HIV-1 infection: good news for an HIV-1 vaccine? Nat. Med. 15, 866–870 (2009).

  16. 16.

    et al. Rational design of envelope identifies broadly neutralizing human monoclonal antibodies to HIV-1. Science 329, 856–861 (2010).

  17. 17.

    et al. Structural basis for broad and potent neutralization of HIV-1 by antibody VRC01. Science 329, 811–817 (2010).

  18. 18.

    et al. Broad and potent neutralizing antibodies from an African donor reveal a new HIV-1 vaccine target. Science 326, 285–289 (2009).

  19. 19.

    et al. Human monoclonal antibody 2G12 defines a distinctive neutralization epitope on the gp120 glycoprotein of human immunodeficiency virus type 1. J. Virol. 70, 1100–1108 (1996).

  20. 20.

    et al. Efficient neutralization of primary isolates of HIV-1 by a recombinant human monoclonal antibody. Science 266, 1024–1027 (1994).

  21. 21.

    et al. Anti-V3 monoclonal antibodies display broad neutralizing activities against multiple HIV-1 subtypes. PLoS ONE 5, e10254 (2010).

  22. 22.

    & Structure-function relationships of HIV-1 envelope sequence-variable regions refocus vaccine design. Nat. Rev. Immunol. 10, 527–535 (2010).

  23. 23.

    et al. A potent cross-clade neutralizing human monoclonal antibody against a novel epitope on gp41 of human immunodeficiency virus type 1. AIDS Res. Hum. Retroviruses 17, 1757–1765 (2001).

  24. 24.

    et al. A conserved neutralizing epitope on gp41 of human immunodeficiency virus type 1. J. Virol. 67, 6642–6647 (1993).

  25. 25.

    et al. Broadly neutralizing antibodies targeted to the membrane-proximal external region of human immunodeficiency virus type 1 glycoprotein gp41. J. Virol. 75, 10892–10905 (2001).

  26. 26.

    et al. Role of HIV membrane in neutralization by two broadly neutralizing antibodies. Proc. Natl. Acad. Sci. USA 106, 20234–20239 (2009).

  27. 27.

    , , , & Epitope mapping of two immunodominant domains of gp41, the transmembrane protein of human immunodeficiency virus type 1, using ten human monoclonal antibodies. J. Virol. 65, 4832–4838 (1991).

  28. 28.

    et al. Neutralization of HIV-1 primary isolates by polyclonal and monoclonal human antibodies. Int. Immunol. 9, 1281–1290 (1997).

  29. 29.

    et al. Nonneutralizing antibodies are able to inhibit human immunodeficiency virus type 1 replication in macrophages and immature dendritic cells. J. Virol. 80, 6177–6181 (2006).

  30. 30.

    , , & Generation of human monoclonal antibodies to human immunodeficiency virus. Proc. Natl. Acad. Sci. USA 86, 1624–1628 (1989).

  31. 31.

    , , , & Effects of oligomerization on the epitopes of the human immunodeficiency virus type 1 envelope glycoproteins. Virology 267, 220–228 (2000).

  32. 32.

    et al. Oligomeric structure of gp41, the transmembrane protein of human immunodeficiency virus type 1. J. Virol. 63, 2674–2679 (1989).

  33. 33.

    & Recognition by human monoclonal antibodies of free and complexed peptides representing the prefusogenic and fusogenic forms of human immunodeficiency virus type 1 gp41. J. Virol. 74, 6186–6192 (2000).

  34. 34.

    et al. Oligomer-specific conformations of the human immunodeficiency virus (HIV-1) gp41 envelope glycoprotein ectodomain recognized by human monoclonal antibodies. AIDS Res. Hum. Retroviruses 25, 319–328 (2009).

  35. 35.

    et al. An affinity-enhanced neutralizing antibody against the membrane-proximal external region of human immunodeficiency virus type 1 gp41 recognizes an epitope between those of 2F5 and 4E10. J. Virol. 81, 4033–4043 (2007).

  36. 36.

    , & Crystal structure of an isoleucine-zipper trimer. Nature 371, 80–83 (1994).

  37. 37.

    et al. Structure of rotavirus outer-layer protein VP7 bound with a neutralizing Fab. Science 324, 1444–1447 (2009).

  38. 38.

    et al. Three-dimensional solution structure of the 44kDa ectodomain of SIV gp41. EMBO J. 17, 4572–4584 (1998).

  39. 39.

    et al. The crystal structure of the SIV gp41 ectodomain at 1.47 A resolution. J. Struct. Biol. 126, 131–144 (1999).

  40. 40.

    et al. Structure and mechanistic analysis of the anti-human immunodeficiency virus type 1 antibody 2F5 in complex with its gp41 epitope. J. Virol. 78, 10724–10737 (2004).

  41. 41.

    et al. Randomized, double-blind, placebo-controlled efficacy trial of a bivalent recombinant glycoprotein 120 HIV-1 vaccine among injection drug users in Bangkok, Thailand. J. Infect. Dis. 194, 1661–1671 (2006).

  42. 42.

    et al. Vaccination with ALVAC and AIDSVAX to prevent HIV-1 infection in Thailand. N. Engl. J. Med. 361, 2209–2220 (2009).

  43. 43.

    et al. Conserved and exposed epitopes on intact, native, primary human immunodeficiency virus type 1 virions of group M. J. Virol. 74, 7096–7107 (2000).

  44. 44.

    et al. Oligomeric and conformational properties of a proteolytically mature, disulfide-stabilized human immunodeficiency virus type 1 gp140 envelope glycoprotein. J. Virol. 76, 7760–7776 (2002).

  45. 45.

    , , , & Dilation of the human immunodeficiency virus-1 envelope glycoprotein fusion pore revealed by the inhibitory action of a synthetic peptide from gp41. J. Cell Biol. 140, 315–323 (1998).

  46. 46.

    & Kinetic dependence to HIV-1 entry inhibition. J. Biol. Chem. 281, 25813–25821 (2006).

  47. 47.

    et al. Nature of nonfunctional envelope proteins on the surface of human immunodeficiency virus type 1. J. Virol. 80, 2515–2528 (2006).

  48. 48.

    & Antibody-dependent cellular cytotoxicity directed against cells expressing human immunodeficiency virus type 1 envelope of primary or laboratory-adapted strains by human and chimpanzee monoclonal antibodies of different epitope specificities. J. Virol. 72, 286–293 (1998).

  49. 49.

    et al. Identification of sites within gp41 that serve as targets for antibody-dependent cellular cytotoxicity by using human monoclonal antibodies. J. Immunol. 145, 3276–3282 (1990).

  50. 50.

    , , , & Functional activities of 20 human immunodeficiency virus type 1 (HIV-1)-specific human monoclonal antibodies. AIDS Res. Hum. Retroviruses 11, 1095–1099 (1995).

  51. 51.

    et al. Anti-gp41 antibodies cloned from HIV-infected patients with broadly neutralizing serologic activity. J. Virol. 84, 5032–5042 (2010).

  52. 52.

    & Molecular replacement with MOLREP. Acta Crystallogr. D Biol. Crystallogr. 66, 22–25 (1997).

  53. 53.

    et al. Phaser crystallographic software. J. Appl. Crystallogr. 40, 658–674 (2007).

  54. 54.

    et al. Toward the structural genomics of complexes: crystal structure of a PE/PPE protein complex from Mycobacterium tuberculosis. Proc. Natl. Acad. Sci. USA 103, 8060–8065 (2006).

  55. 55.

    & Electron-density map interpretation. Methods Enzymol. 277, 173–208 (1997).

  56. 56.

    , , & Features and development of Coot. Acta Crystallogr. D Biol. Crystallogr. 66, 486–501 (2010).

  57. 57.

    et al. PHENIX: a comprehensive Python-based system for macromolecular structure solution. Acta Crystallogr. D Biol. Crystallogr. 66, 213–221 (2010).

  58. 58.

    Refinement of macromolecular structures by the maximum-likelihood method. Acta Crystallogr. D Biol. Crystallogr. 53, 240–255 (1997).

  59. 59.

    , , & PROCHECK: A program to check the stereochemical quality of protein structures. J. Appl. Crystallogr. 26, 283–291 (1993).

  60. 60.

    The PyMOL User's Manual (DeLano Scientific, San Carlos, California, USA, 2002).

Download references

Acknowledgements

We thank S. Harrison, H. Peng, A. Carfi, A. Dey, J. Mascola, M. Alam and B.F. Haynes for advice and assistance, and the staff of the Northeastern Collaborative Access Team at Advanced Photon Source, Argonne National Laboratory, for assistance with X-ray data collection. We acknowledge support from US National Institutes of Health grants GM083680 (to B.C.), AI084794 (to B.C. and Dan H. Barouch) and AI36085 (to S.Z.-P.); a Collaboration for AIDS Vaccine Discovery grant (to Barton F. Haynes) from the Bill and Melinda Gates Foundation; the Center for HIV/AIDS Vaccine Immunology (to Barton F. Haynes); and the Department of Veterans Affairs. J.C. is supported by a fellowship from the Ragon Institute of MGH, MIT and Harvard.

Author information

Author notes

    • Gary Frey
    •  & Jia Chen

    These authors contributed equally to this work.

Affiliations

  1. Division of Molecular Medicine, Children's Hospital, Harvard Medical School, Boston, Massachusetts, USA.

    • Gary Frey
    • , Jia Chen
    • , Sophia Rits-Volloch
    • , Michael M Freeman
    •  & Bing Chen
  2. Department of Pediatrics, Harvard Medical School, Boston, Massachusetts, USA.

    • Gary Frey
    • , Jia Chen
    • , Sophia Rits-Volloch
    • , Michael M Freeman
    •  & Bing Chen
  3. Department of Pathology, New York University School of Medicine, New York, New York, USA.

    • Susan Zolla-Pazner
  4. New York Veterans Affairs Medical Center, New York, New York, USA.

    • Susan Zolla-Pazner

Authors

  1. Search for Gary Frey in:

  2. Search for Jia Chen in:

  3. Search for Sophia Rits-Volloch in:

  4. Search for Michael M Freeman in:

  5. Search for Susan Zolla-Pazner in:

  6. Search for Bing Chen in:

Contributions

G.F., J.C. and B.C. designed research; G.F., J.C., S.R.-V. and M.M.F. performed the experiments; S.Z.-P. provided antibodies; G.F., J.C., S.R.-V., M.M.F. and B.C. analyzed data; and G.F., J.C., S.Z.-P. and B.C. wrote the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Bing Chen.

Supplementary information

PDF files

  1. 1.

    Supplementary Text and Figures

    Supplementary Figures 1–5, Supplementary Table 1 and Supplementary Methods

About this article

Publication history

Received

Accepted

Published

DOI

https://doi.org/10.1038/nsmb.1950

Further reading