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HCV Broadly Neutralizing Antibodies Use a CDRH3 Disulfide Motif to Recognize an E2 Glycoprotein Site that Can Be Targeted for Vaccine Design

Flyak, Andrew I. and Ruiz, Stormy and Colbert, Michelle D. and Luong, Tiffany and Crowe, James E., Jr. and Bailey, Justin R. and Bjorkman, Pamela J. (2018) HCV Broadly Neutralizing Antibodies Use a CDRH3 Disulfide Motif to Recognize an E2 Glycoprotein Site that Can Be Targeted for Vaccine Design. Cell Host & Microbe, 24 (5). pp. 703-716. ISSN 1931-3128. PMCID PMC6258177. doi:10.1016/j.chom.2018.10.009.

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Hepatitis C virus (HCV) vaccine efforts are hampered by the extensive genetic diversity of HCV envelope glycoproteins E1 and E2. Structures of broadly neutralizing antibodies (bNAbs) (e.g., HEPC3, HEPC74) isolated from individuals who spontaneously cleared HCV infection facilitate immunogen design to elicit antibodies against multiple HCV variants. However, challenges in expressing HCV glycoproteins previously limited bNAb-HCV structures to complexes with truncated E2 cores. Here we describe crystal structures of full-length E2 ectodomain complexes with HEPC3 and HEPC74, revealing lock-and-key antibody-antigen interactions, E2 regions (including a target of immunogen design) that were truncated or disordered in E2 cores, and an antibody CDRH3 disulfide motif that exhibits common interactions with a conserved epitope despite different bNAb-E2 binding orientations. The structures display unusual features relevant to common genetic signatures of HCV bNAbs and demonstrate extraordinary plasticity in antibody-antigen interactions. In addition, E2 variants that bind HEPC3/HEPC74-like germline precursors may represent candidate vaccine immunogens.

Item Type:Article
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URLURL TypeDescription CentralArticle
Flyak, Andrew I.0000-0002-8722-479X
Crowe, James E., Jr.0000-0002-0049-1079
Bailey, Justin R.0000-0001-5704-3130
Bjorkman, Pamela J.0000-0002-2277-3990
Alternate Title:Broadly neutralizing antibodies against HCV use a CDRH3 disulfide motif to recognize an E2 glycoprotein site that can be targeted for vaccine design
Additional Information:© 2018 Elsevier. Received 3 July 2018, Revised 17 August 2018, Accepted 3 October 2018, Available online 14 November 2018. We thank the Caltech Protein Expression Center for help with protein expression and Jens Kaiser, Christopher Barnes, Beth Stadtmueller, and Harry Gristick for training in crystallography. This research was supported by National Institutes of Health grant R01 AI127469 (to J.R.B. and P.J.B.), U19 AI088791 (to J.R.B.) (content is solely the responsibility of the authors and does not necessarily represent the official views of the NIH) and the Molecular Observatory at Caltech supported by the Gordon and Betty Moore Foundation. A.I.F. is a Cancer Research Institute Irvington Fellow supported by the Cancer Research Institute. This research used resources of the Advanced Photon Source, Advanced Light Source, and Stanford Synchrotron Radiation Lightsource, SLAC National Accelerator Laboratory. Use of the Advanced Photon Source was supported by the U.S. Department of Energy, Office of Science, Office of Basic Energy Sciences, under Contract No. DE-AC02-06CH11357. The Advanced Light Source is a DOE Office of Science User Facility under contract no. DE-AC02-05CH11231. Use of the Stanford Synchrotron Radiation Lightsource, SLAC National Accelerator Laboratory, is supported by the U.S. Department of Energy, Office of Science, Office of Basic Energy Sciences under Contract No. DE-AC02-76SF00515. The SSRL Structural Molecular Biology Program is supported by the DOE Office of Biological and Environmental Research and by NIHGMSP41GM103393. Author Contributions: A.I.F., J.R.B., and P.J.B. conceived the study; A.I.F. and S.R. optimized the crystallography conditions; A.I.F. solved and analyzed the crystal structures; T.L. expressed and purified proteins; S.R. performed ELISA experiments; M.D.C. performed neutralization experiments; J.E.C. provided antibody sequences; A.I.F., S.R., M.D.C., J.R.B., J.E.C., and P.J.B. analyzed the data; A.I.F. and P.J.B. wrote the original draft; all authors reviewed and edited the manuscript. Data and Software Availability: Coordinates for atomic models are deposited in the Protein Data Bank under the accession numbers PDB: 6MED (HEPC3), PDB: 6MEE (HEPC74), PDB: 6MEF (AR3C), PDB: 6MEG (HEPC46), PDB: 6MEH (HEPC74-E2ecto), PDB: 6MEI (HEPC3-E2ecto), PDB: 6MEJ (HEPC3-E2ecto-HEPC46), and PDB: 6MEK (HEPC3-E2core-HEPC46). Declaration of Interests: A.I.F., J.E.C., and J.R.B. are inventors of patents submitted pertaining to some of the antibodies and antigens presented in this paper. J.E.C. has served as a consultant for Takeda Vaccines, Sanofi Pasteur, Pfizer, and Novavax; is on the Scientific Advisory Boards of CompuVax, GigaGen, Meissa Vaccines, and PaxVax; and is Founder of IDBiologics. The other authors declare no competing interests.
Funding AgencyGrant Number
NIHR01 AI127469
NIHU19 AI088791
Gordon and Betty Moore FoundationUNSPECIFIED
Cancer Research InstituteUNSPECIFIED
Department of Energy (DOE)DE-AC02-06CH11357
Department of Energy (DOE)DE-AC02-05CH11231
Department of Energy (DOE)DE-AC02-76SF00515
Subject Keywords:broadly neutralizing antibodies; structure; HCV; E2; glycoprotein; epitope; disulfide bond
Issue or Number:5
PubMed Central ID:PMC6258177
Record Number:CaltechAUTHORS:20181114-153017863
Persistent URL:
Official Citation:Andrew I. Flyak, Stormy Ruiz, Michelle D. Colbert, Tiffany Luong, James E. Crowe, Justin R. Bailey, Pamela J. Bjorkman, HCV Broadly Neutralizing Antibodies Use a CDRH3 Disulfide Motif to Recognize an E2 Glycoprotein Site that Can Be Targeted for Vaccine Design, Cell Host & Microbe, Volume 24, Issue 5, 2018, Pages 703-716.e3, ISSN 1931-3128,
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:90898
Deposited By: George Porter
Deposited On:15 Nov 2018 00:18
Last Modified:16 Feb 2022 23:51

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