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Synthetic glycopeptides reveal the glycan specificity of HIV-neutralizing antibodies

Nature Chemical Biology volume 9, pages 521526 (2013) | Download Citation

Abstract

A new class of glycan-reactive HIV-neutralizing antibodies, including PG9 and PG16, has been recently discovered that seem to recognize previously uncharacterized glycopeptide epitopes on HIV-1 gp120. However, further characterization and reconstitution of the precise neutralizing epitopes are complicated by the heterogeneity of glycosylation. We report here the design, synthesis and antigenic evaluation of new cyclic V1V2 glycopeptides carrying defined N-linked glycans at the conserved glycosylation sites (Asn160 and Asn156 or Asn173) derived from gp120 of two HIV-1 isolates. Antibody binding studies confirmed the necessity of a Man5GlcNAc2 glycan at Asn160 for recognition by PG9 and PG16 and further revealed a critical role of a sialylated N-glycan at the secondary site (Asn156 or Asn173) in the context of glycopeptides for antibody binding. In addition to defining the glycan specificities of PG9 and PG16, the identified synthetic glycopeptides provide a valuable template for HIV-1 vaccine design.

  • Compound C38H63NO30

    2-Methyl-[α-D-mannopyranosyl-(1-3)-[α-D-mannopyranosyl-(1-6)-]-α-D-mannopyranosyl-(1-6)-[α-D-mannopyranosyl-(1-3)-]-β-D-mannopyranosyl-(1-4)-1,2-dideoxy-α-D-glucopyrano]-[2,1-d]-oxazoline

  • Compound C62H103NO50

    2-Methyl-{α-D-mannopyranosyl-(1-2)-α-D-mannopyranosyl-(1-6)-[α-D-mannopyranosyl-(1-2)-[α-D-mannopyranosyl-(1-3)]-α-D-mannopyranosyl-(1-6)-[α-D-mannopyranosyl-(1-2)-α-D-mannopyranosyl-(1-2)-α-D-mannopyranosyl-(1-3)-]-β-D-mannopyranosyl-(1-4)-1,2-dideoxy-α-D-glucopyrano}-[2,1-d]-oxazoline

  • Compound C76H123N5O56

    2-Methyl-{(5-acetamido-3,5-dideoxy-D-glycero-α-D-galacto-2-nonulopyranosylonic acid)-(2-6)-β-D-galactopyranosyl-(1-4)-2-acetamido-2-deoxy-β-D-glucopyranosyl-(1-2)-α-D-mannopyranosyl-(1-6)-[(5-acetamido-3,5-dideoxy-D-glycero-α-D-galacto-2-nonulopyranosylonic acid)-(2-6)-β-D-galactopyranosyl-(1-4)-2-acetamido-2-deoxy-β-D-glucopyranosyl-(1-2)-α-D-mannopyranosyl-(1-3)-]-β-D-mannopyranosyl-(1-4)-1,2-dideoxy-α-D-glucopyrano}-[2,1-d]-oxazoline

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Acknowledgements

We thank S. Fan (University of Maryland) for providing the recombinant Endo-D and K. Moremen and Y. Xiang (University of Georgia) for providing the recombinant mouse α-1,2-mannosidase. This work is supported in parts by grants from the National Institute of Allergy and Infectious Diseases (NIAID) (US National Institutes of Health (NIH) grant 1R21AI101035 to L.-X.W.), the International AIDS Vaccine Initiative's Neutralizing Antibody Consortium and by the Intramural Research Program of the Vaccine Research Center, NIAID-NIH.

Author information

Affiliations

  1. Institute of Human Virology, University of Maryland School of Medicine, Baltimore, Maryland, USA.

    • Mohammed N Amin
    • , Wei Huang
    • , Jared Orwenyo
    •  & Lai-Xi Wang
  2. Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine, Baltimore, Maryland, USA.

    • Mohammed N Amin
    • , Wei Huang
    • , Jared Orwenyo
    •  & Lai-Xi Wang
  3. Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland, USA.

    • Jason S McLellan
    •  & Peter D Kwong
  4. Department of Immunology and Microbial Science, International AIDS Vaccine Initiative (IAVI) Neutralizing Antibody Center and Center for HIV/AIDS Vaccine Immunology and Immunogen Discovery, The Scripps Research Institute, La Jolla, California, USA.

    • Dennis R Burton
  5. Ragon Institute of Massachusetts General Hospital, Massachusetts Institute of Technology and Harvard, Cambridge, Massachusetts, USA.

    • Dennis R Burton
  6. IAVI, New York, New York, USA.

    • Wayne C Koff

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Contributions

M.N.A., J.S.M., W.H., P.D.K. and L.-X.W. designed the research and analyzed the data; M.N.A., J.S.M., W.H. and J.O. performed the research; L.-X.W. conceived the idea and supervised the research; D.R.B. and W.C.K. contributed PG9 and PG16 antibodies; L.-X.W. and M.N.A. wrote the manuscript; all of the authors contributed to revisions of the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Lai-Xi Wang.

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DOI

https://doi.org/10.1038/nchembio.1288

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