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Title: Inhibition of EBV-mediated membrane fusion by anti-gHgL antibodies

Abstract

Herpesvirus entry into cells requires the coordinated action of multiple virus envelope glycoproteins, including gH, gL, and gB. For EBV, the gp42 protein assembles into complexes with gHgL heterodimers and binds HLA class II to activate gB-mediated membrane fusion with B cells. EBV tropism is dictated by gp42 levels in the virion, as it inhibits entry into epithelial cells while promoting entry into B cells. The gHgL and gB proteins are targets of neutralizing antibodies and potential candidates for subunit vaccine development, but our understanding of their neutralizing epitopes and the mechanisms of inhibition remain relatively unexplored. Here we studied the structures and mechanisms of two anti-gHgL antibodies, CL40 and CL59, that block membrane fusion with both B cells and epithelial cells. We determined the structures of the CL40 and CL59 complexes with gHgL using X-ray crystallography and EM to identify their epitope locations. CL59 binds to the C-terminal domain IV of gH, while CL40 binds to a site occupied by the gp42 receptor binding domain. CL40 binding to gHgL/gp42 complexes is not blocked by gp42 and does not interfere with gp42 binding to HLA class II, indicating that its ability to block membrane fusion with B cells represents amore » defect in gB activation. Furthermore, these data indicate that anti-gHgL neutralizing antibodies can block gHgL-mediated activation of gB through different surface epitopes and mechanisms.« less

Authors:
ORCiD logo [1];  [2];  [3];  [4];  [2];  [4];  [1]
+ Show Author Affiliations
  1. Stanford Univ. School of Medicine, Stanford, CA (United States)
  2. Univ. of California, Los Angeles, CA (United States)
  3. Northwestern Univ., Chicago, IL (United States); Technische Univ. Munchen, Munich (Germany)
  4. Northwestern Univ., Chicago, IL (United States)
Publication Date:
Research Org.:
SLAC National Accelerator Laboratory (SLAC), Menlo Park, CA (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1439107
Grant/Contract Number:  
AC02-76SF00515; AI119480; AI076183; CA117794
Resource Type:
Accepted Manuscript
Journal Name:
Proceedings of the National Academy of Sciences of the United States of America
Additional Journal Information:
Journal Volume: 114; Journal Issue: 41; Journal ID: ISSN 0027-8424
Publisher:
National Academy of Sciences
Country of Publication:
United States
Language:
English
Subject:
70 PLASMA PHYSICS AND FUSION TECHNOLOGY; herpesvirus; EBV; membrane fusion; neutralizing antibody; glycoprotein complex

Citation Formats

Sathiyamoorthy, Karthik, Jiang, Jiansen, Mohl, Britta S., Chen, Jia, Zhou, Z. Hong, Longnecker, Richard, and Jardetzky, Theodore S. Inhibition of EBV-mediated membrane fusion by anti-gHgL antibodies. United States: N. p., 2017. Web. doi:10.1073/pnas.1704661114.
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Sathiyamoorthy, Karthik, Jiang, Jiansen, Mohl, Britta S., Chen, Jia, Zhou, Z. Hong, Longnecker, Richard, & Jardetzky, Theodore S. Inhibition of EBV-mediated membrane fusion by anti-gHgL antibodies. United States. https://doi.org/10.1073/pnas.1704661114
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Sathiyamoorthy, Karthik, Jiang, Jiansen, Mohl, Britta S., Chen, Jia, Zhou, Z. Hong, Longnecker, Richard, and Jardetzky, Theodore S. Fri . "Inhibition of EBV-mediated membrane fusion by anti-gHgL antibodies". United States. https://doi.org/10.1073/pnas.1704661114. https://www.osti.gov/servlets/purl/1439107.
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@article{osti_1439107,
title = {Inhibition of EBV-mediated membrane fusion by anti-gHgL antibodies},
author = {Sathiyamoorthy, Karthik and Jiang, Jiansen and Mohl, Britta S. and Chen, Jia and Zhou, Z. Hong and Longnecker, Richard and Jardetzky, Theodore S.},
abstractNote = {Herpesvirus entry into cells requires the coordinated action of multiple virus envelope glycoproteins, including gH, gL, and gB. For EBV, the gp42 protein assembles into complexes with gHgL heterodimers and binds HLA class II to activate gB-mediated membrane fusion with B cells. EBV tropism is dictated by gp42 levels in the virion, as it inhibits entry into epithelial cells while promoting entry into B cells. The gHgL and gB proteins are targets of neutralizing antibodies and potential candidates for subunit vaccine development, but our understanding of their neutralizing epitopes and the mechanisms of inhibition remain relatively unexplored. Here we studied the structures and mechanisms of two anti-gHgL antibodies, CL40 and CL59, that block membrane fusion with both B cells and epithelial cells. We determined the structures of the CL40 and CL59 complexes with gHgL using X-ray crystallography and EM to identify their epitope locations. CL59 binds to the C-terminal domain IV of gH, while CL40 binds to a site occupied by the gp42 receptor binding domain. CL40 binding to gHgL/gp42 complexes is not blocked by gp42 and does not interfere with gp42 binding to HLA class II, indicating that its ability to block membrane fusion with B cells represents a defect in gB activation. Furthermore, these data indicate that anti-gHgL neutralizing antibodies can block gHgL-mediated activation of gB through different surface epitopes and mechanisms.},
doi = {10.1073/pnas.1704661114},
journal = {Proceedings of the National Academy of Sciences of the United States of America},
number = 41,
volume = 114,
place = {United States},
year = {Fri Sep 22 00:00:00 EDT 2017},
month = {Fri Sep 22 00:00:00 EDT 2017}
}
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Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record
https://doi.org/10.1073/pnas.1704661114
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Cited by: 21 works
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Figures / Tables:

Fig. 1 Fig. 1: CL40 and CL59 inhibition of membrane fusion. (A and B) Inhibition of fusion activity with purified CL40 mAb (lighter color shade) and Fab (darker color shade) using epithelial cells (A) and B cells (B). (C and D) Inhibition of fusion activity with purified CL59 mAb (lighter color shade)more » and Fab (darker color shade) using epithelial cells (C) and B cells (D). For these panels, the x-axis indicates the amount of purified antibody as final concentration in nanomoles. Empty pSG5 plasmid with no gH or gL insert serves as the negative control, and fusion activity with no antibody is set to 100% for HEK 293 epithelial cells (maroon) and Daudi B cells (blue). Each data point is represented by a single experiment titrating the antibody (except negative control). (E and F) Fusion activity expressed as the average from three independent experiments (biological replicates) with a 200 nM final antibody concentration, with fusion activity expressed as percentage with WT and no antibody data point set at 100% for HEK 293 epithelial cells (E) and Daudi B cells (F). In each experiment, the negative control is pSG5 plasmid with no gH or gL insert.« less
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