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Rational design of a CD4 mimic that inhibits HIV-1 entry and exposes cryptic neutralization epitopes

Nature Biotechnology volume 21, pages 71–76 (2003)Cite this article

Abstract

The conserved surfaces of the human immunodeficiency virus (HIV)-1 envelope involved in receptor binding represent potential targets for the development of entry inhibitors and neutralizing antibodies. Using structural information on a CD4-gp120-17b antibody complex, we have designed a 27-amino acid CD4 mimic, CD4M33, that presents optimal interactions with gp120 and binds to viral particles and diverse HIV-1 envelopes with CD4-like affinity. This mini-CD4 inhibits infection of both immortalized and primary cells by HIV-1, including primary patient isolates that are generally resistant to inhibition by soluble CD4. Furthermore, CD4M33 possesses functional properties of CD4, including the ability to unmask conserved neutralization epitopes of gp120 that are cryptic on the unbound glycoprotein. CD4M33 is a prototype of inhibitors of HIV-1 entry and, in complex with envelope proteins, a potential component of vaccine formulations, or a molecular target in phage display technology to develop broad-spectrum neutralizing antibodies.

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Figure 1: Structure, sequence, and computed complex of CD4 miniprotein.
Figure 2: Binding activity of the CD4M33 miniprotein.
Figure 3: Conformational changes in gp120 and HIV-1 envelope induced by CD4M33 binding.

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Acknowledgements

We thank N. Schulke (Progenics, Tarrytown, NY) for four-domain sCD4 and gp120 from R5-tropic JRFL; I. Srivastava (Chiron Co., Emeryville, CA) for recombinant gp120 from X4-tropic SF2; S. Leow (Pharmacia, Kalamazoo, MI) for two-domain sCD4; M. Parmentier (University of Brussels, Belgium) for CHO-K1 CCR5 cell line. We thank E.A. Berger (Laboratory of Viral Diseases (LVD), NIH, Bethesda, MD) for providing vaccinia virus constructs, G. Scarlatti (Unit of Viral Biology and Transmission, San Raffaele Scientific Institute, Milan, Italy) for providing primary HIV-1 isolates, and O. Combes, D. Seiller (Commissariat Energie Atomique (CEA), Saclay, France), M. Cerutti, M. Secondy (Institut Recherche Developpement (IRD), Montpellier, France), for their assistance and comments. We thank the late J.C. Mani for his assistance and discussion in optical biosensor experiments. We acknowledge the Centralised Facility for AIDS Reagents, NIBSC, UK (EU Programme EVA/MRC, Grant Nr QLK2-CT-1999-00609 and GP828102) for recombinant gp120 from R5 W61D, anti-human CD4 L120.3 and 17b, 48d monoclonal antibody. We thank the French Agence Nationale de Recherches sur le SIDA (ANRS) and Ensemble Contre le SIDA—SIDACTION for financial support.

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Authors and Affiliations

  1. Department of Protein Engineering and Research, CEA Saclay, Gif-sur-Yvette, 91191, France

    Loïc Martin, François Stricher, Isabelle Freulon, André Ménez & Claudio Vita

  2. Retroviral and Molecular Immunology Laboratory, IRD/CNRS, Montpellier, 34094, France

    Dorothée Missé, Xavier Magne & Francisco Veas

  3. Unit of Human Virology, San Raffaele Scientific Institute, Milan, 20132, Italy

    Francesca Sironi & Paolo Lusso

  4. Biotechnology and Pharmacology Institute, Faculty of Pharmacy, Montpellier, 34093, France

    Martine Pugnière

  5. Structural Biochemistry Center, Faculty of Pharmacy, Montpellier, 34093, France

    Philippe Barthe, Rafael Prado-Gotor & Christian Roumestand

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Correspondence to Claudio Vita.

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Martin, L., Stricher, F., Missé, D. et al. Rational design of a CD4 mimic that inhibits HIV-1 entry and exposes cryptic neutralization epitopes. Nat Biotechnol 21, 71–76 (2003). https://doi.org/10.1038/nbt768

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