1. A gp41 MPER-specific Llama VHH Requires a Hydrophobic CDR3 for Neutralization but not for Antigen Recognition
- Author
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Laura E. McCoy, Simone Battella, Charles Sabin, Alexandre M. J. J. Bonvin, Andreas Hinz, Robin A. Weiss, Michael S. Seaman, David Lutje Hulsik, Lucy Rutten, Miriam Hock, C. Theo Verrips, Alejandra Ramos, Winfried Weissenhorn, Nika M. Strokappe, Pauline Macheboeuf, Johannes P. M. Langedijk, Mohamed El Khattabi, Adrien Favier, Pascal Poignard, Ying-ying Liu, David Davis, Marlén M. I. Aasa-Chapman, Jean-Pierre Simorre, Anna Forsman Quigley, Unit for Virus Host-Cell Interactions [Grenoble] (UVHCI), Université Joseph Fourier - Grenoble 1 (UJF)-European Molecular Biology Laboratory [Grenoble] (EMBL)-Centre National de la Recherche Scientifique (CNRS), Departments of Applied Physics [New Haven], Yale University [New Haven], Research Department of Infection and Population Health [London], University College of London [London] (UCL), Independent Department for Medical Psychology and Medical Sociology, Universität Leipzig, Beth Israel Deaconess Medical Center [Boston] (BIDMC), Harvard Medical School [Boston] (HMS), Centre d'Immunologie de Marseille - Luminy (CIML), Aix Marseille Université (AMU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Institut de biologie structurale (IBS - UMR 5075 ), Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), ANR-10-LABX-0049,GRAL,Grenoble Alliance for Integrated Structural Cell Biology(2010), Centre National de la Recherche Scientifique (CNRS)-European Molecular Biology Laboratory [Grenoble] (EMBL)-Université Joseph Fourier - Grenoble 1 (UJF), Universität Leipzig [Leipzig], Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)
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Macromolecular Assemblies ,MESH: HIV Envelope Protein gp41 ,MESH: Amino Acid Sequence ,Complementarity determining region ,Adaptive Immunity ,HIV Antibodies ,MESH: Base Sequence ,Antibody production ,Neutralization ,Epitope ,MESH: Antibodies, Neutralizing ,Epitopes ,Immunodeficiency Viruses ,Bacteriophages ,MESH: Animals ,Biomacromolecule-Ligand Interactions ,Enzyme-linked immunoassays ,lcsh:QH301-705.5 ,chemistry.chemical_classification ,0303 health sciences ,[SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM] ,MESH: Hydrophobic and Hydrophilic Interactions ,MESH: Neutralization Tests ,030302 biochemistry & molecular biology ,Antivirals ,Immunizations ,Lipids ,HIV Envelope Protein gp41 ,MESH: Surface Plasmon Resonance ,3. Good health ,MESH: Mutagenesis, Site-Directed ,International (English) ,Viral Envelope ,MESH: Camelids, New World ,MESH: Complementarity Determining Regions ,MESH: Immunization ,Antibody ,Camelids, New World ,Hydrophobic and Hydrophilic Interactions ,Research Article ,MESH: Proteolipids ,lcsh:Immunologic diseases. Allergy ,MESH: Epitopes ,medicine.drug_class ,Proteolipids ,Molecular Sequence Data ,Immunology ,Biophysics ,Membrane fusion ,Viral Structure ,Biology ,Monoclonal antibody ,Gp41 ,MESH: Single-Domain Antibodies ,Microbiology ,Antibodies ,Cell Line ,03 medical and health sciences ,Viral envelope ,Neutralization Tests ,ddc:570 ,Virology ,Genetics ,medicine ,Animals ,Humans ,Amino Acid Sequence ,Microbial Pathogens ,Molecular Biology ,030304 developmental biology ,MESH: Humans ,MESH: Molecular Sequence Data ,Base Sequence ,MESH: HIV Antibodies ,Crystal structure ,Immunity ,Viral Vaccines ,Single-Domain Antibodies ,Surface Plasmon Resonance ,Antibodies, Neutralizing ,Complementarity Determining Regions ,Molecular biology ,MESH: Cell Line ,lcsh:Biology (General) ,chemistry ,Mutagenesis, Site-Directed ,HIV-1 ,biology.protein ,Immunization ,Parasitology ,lcsh:RC581-607 ,Glycoprotein - Abstract
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., Author Summary Due to the absence of an effective vaccine or cure for acquired immunodeficiency syndrome (AIDS), HIV-1 infections still result in high mortality. Two antibodies, 2F5 and 4E10, previously isolated from HIV-1 infected patients, prevent infections by binding to the MPER of gp41, a part of the virus that is difficult to access and only transiently exposed. Here, we immunized llamas with a gp41-based immunogen and subsequently isolated a small antibody fragment (VHH) that can easily access and recognize the MPER. We showed that a unit of two VHH, named bi-2H10, was indeed capable of preventing HIV-1 from infecting cells. We determined the three dimensional structure of the VHH and mapped its interaction site to an MPER region that overlaps with the 2F5 epitope. The 2H10 VHH displays a membrane binding component important for neutralization that resembles that of 2F5. In conclusion, we have developed an immunogen and a small antibody that may have great potential for development of novel anti-HIV/AIDS vaccines and treatments.
- Published
- 2013
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