A gp41 MPER-specific Llama VHH Requires a Hydrophobic CDR3 for Neutralization but not for Antigen Recognition

PLoS Pathogens

Alexandre M. J. J. Bonvin¹  Nika M. Strokappe²  Ying-ying Liu²  Simone Battella²  Mohamed El Khattabi²  Robin A. Weiss³  Lucy Rutten³  Winfried Weissenhorn⁴  Jean-Pierre Simorre⁴  Pascal Poignard⁵  Alejandra Ramos⁵  David Davis⁶  Michael S. Seaman⁷  Adrien Favier⁸  Laura E. McCoy⁹  Anna Forsman Quigley⁹  Marlén M. I. Aasa-Chapman⁹  Johannes P. M. Langedijk^1,10  C. Theo Verrips^1,11  David Lutje Hulsik^1,12  Andreas Hinz^1,12  Miriam Hock^1,12  Pauline Macheboeuf^1,12  Charles Sabin^1,12 

[1] Bijvoet Center for Biomolecular Research, Faculty of Science, Utrecht University, Utrecht, The Netherlands;Biomolecular Imaging (BMI), Faculty of Science, Utrecht University, Utrecht, The Netherlands;CEA, Institut de Biologie Structurale-Jean-Pierre Ebel, Grenoble Cedex, France;CNRS, Institut de Biologie Structurale-Jean-Pierre Ebel, Grenoble Cedex, France;Department of Immunology and Microbial Science, International AIDS Vaccine Initiative Neutralizing Antibody Center, The Scripps Research Institute, La Jolla, California, United States of America;Department of Virology, Biomedical Primate Research Centre (BPRC), Rijswijk, The Netherlands;Division of Viral Pathogenesis, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts, United States of America;International AIDS Vaccine Initiative, New York, New York, United States of America;MRC/UCL Centre for Medical Molecular Virology, Division of Infection and Immunity, University College London, London, United Kingdom;Pepscan Therapeutics BV, Lelystad, The Netherlands;UJF-Grenoble-1, Institut de Biologie Structurale-Jean-Pierre Ebel, Grenoble Cedex, France;Unit of Virus Host Cell Interactions (UVHCI), UMI 3265, Université Joseph Fourier-EMBL-CNRS, Grenoble, France

关键词: Antibodies;    HIV-1;    Bacteriophages;    Crystal structure;    Lipids;    Enzyme-linked immunoassays;    Membrane fusion;    Antibody production;   

DOI  :  10.1371/journal.ppat.1003202

学科分类：生物科学（综合）

来源: Public Library of Science

【 摘 要 】

The membrane proximal external region (MPER) of the HIV-1 glycoprotein gp41 is targeted by the broadly neutralizing antibodies 2F5 and 4E10. To date, no immunization regimen in animals or humans has produced HIV-1 neutralizing MPER-specific antibodies. We immunized llamas with gp41-MPER proteoliposomes and selected a MPER-specific single chain antibody (VHH), 2H10, whose epitope overlaps with that of mAb 2F5. Bi-2H10, a bivalent form of 2H10, which displayed an approximately 20-fold increased affinity compared to the monovalent 2H10, neutralized various sensitive and resistant HIV-1 strains, as well as SHIV strains in TZM-bl cells. X-ray and NMR analyses combined with mutagenesis and modeling revealed that 2H10 recognizes its gp41 epitope in a helical conformation. Notably, tryptophan 100 at the tip of the long CDR3 is not required for gp41 interaction but essential for neutralization. Thus bi-2H10 is an anti-MPER antibody generated by immunization that requires hydrophobic CDR3 determinants in addition to epitope recognition for neutralization similar to the mode of neutralization employed by mAbs 2F5 and 4E10.

【 授权许可】

CC BY