Published April 13, 2021
| Published + Supplemental Material + Submitted
Journal Article
Open
Convergence of a common solution for broad ebolavirus neutralization by glycan cap-directed human antibodies
- Creators
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Murin, Charles D.
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Gilchuk, Pavlo
- Ilinykh, Philipp A.
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Huang, Kai
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Kuzmina, Natalia
- Shen, Xiaoli
- Brunh, Jessica F.
- Bryan, Aubrey L.
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Davidson, Edgar
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Doranz, Benjamin J.
- Williamson, Lauren E.
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Copps, Jeffrey
- Alkutkar, Tanwee
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Flyak, Andrew I.
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Bukreyev, Alexander
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Crowe, James E., Jr.
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Ward, Andrew B.
Abstract
Antibodies that target the glycan cap epitope on the ebolavirus glycoprotein (GP) are common in the adaptive response of survivors. A subset is known to be broadly neutralizing, but the details of their epitopes and basis for neutralization are not well understood. Here, we present cryoelectron microscopy (cryo-EM) structures of diverse glycan cap antibodies that variably synergize with GP base-binding antibodies. These structures describe a conserved site of vulnerability that anchors the mucin-like domains (MLDs) to the glycan cap, which we call the MLD anchor and cradle. Antibodies that bind to the MLD cradle share common features, including use of IGHV1-69 and IGHJ6 germline genes, which exploit hydrophobic residues and form β-hairpin structures to mimic the MLD anchor, disrupt MLD attachment, destabilize GP quaternary structure, and block cleavage events required for receptor binding. Our results provide a molecular basis for ebolavirus neutralization by broadly reactive glycan cap antibodies.
Additional Information
© 2021 The Author(s). This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). Received 4 November 2020, Revised 19 February 2021, Accepted 23 March 2021, Available online 13 April 2021. This work was supported by NIH grants U19 AI109762 and U19 AI142785. T.A. is supported by a Kellogg Graduate Student Fellowship from Scripps Research. We would like to thank the Joint Center for Structural Genomics at Scripps Research and Henry Tien for assistance with setting up crystal trays. We would like to thank Dr. Robyn Stanfield for assistance with looping and shipping crystals, collecting X-ray diffraction data, reducing the data, and phasing the crystal structure. We also thank Dr. Ian Wilson for generously sharing synchrotron time to collect X-ray diffraction data. Use of the Stanford Synchrotron Radiation Lightsource, SLAC National Accelerator Laboratory, is supported by the US Department of Energy, Office of Science, Office of Basic Energy Sciences under contract DE-AC02-76SF00515. The SSRL Structural Molecular Biology Program is supported by the DOE Office of Biological and Environmental Research and the National Institutes of Health, National Institute of General Medical Sciences (including P41GM103393). The contents of this publication are solely the responsibility of the authors and do not necessarily represent the official views of NIGMS or NIH. The Jurkat-EBOV GP cell line was a kind gift form Carl Davis and Rafi Ahmed. We thank Hannah Turner, Bill Anderson, Jonathan Torres, Gabriel Ozorowski, and Charles Bowman from Scripps Research for assistance with cryo-EM sample preparation, microscope operation, and data collection and processing. Author contributions: C.D.M. performed protein production for all cryo-EM and kinetic experiments, performed cryo-EM experiments and analyses, crystallized the BDBV289 Fab, performed kinetic experiments and analyses, and collected the crystal data and phased the data. J.F.B. built and validated the crystal structure. J.C. produced Fab for the crystallography trials. P.G. performed synergy, GP cleavage, sGP binding, and mAb autoreactivity assays. T.A. helped to perform GP stability assays and collect cryo-EM data. L.E.W. expressed and purified EBOV-237 and BDBV-329 Fab. P.A.I., K.H., N.K., X.S., A.I.F., and A.B. isolated and characterized EBOV-293 and EBOV-296. A.L.B., E.D., and B.J.D. performed alanine scanning and characterization of EBOV-293 and EBOV-296. C.D.M., P.G., A.L.B., J.E.C., and A.B.W. designed the experiments. C.D.M., P.G., A.B., J.E.C., and A.B.W. wrote and edited the manuscript. Declaration of interests: A.L.B., E.D., and B.J.D. are employees of Integral Molecular. B.J.D. is a shareholder of Integral Molecular. J.E.C. has served as a consultant for Lilly and Luna Biologics, is a member of the Scientific Advisory Boards of CompuVax and Meissa Vaccines, and is the founder of IDBiologics. The Crowe laboratory at Vanderbilt University Medical Center has received sponsored research agreements from and IDBiologics and AstraZeneca. Vanderbilt University has applied for a patent that is related to antibodies discussed in this work. All other authors declare no competing interests. Inclusion and diversity: One or more of the authors of this paper self-identifies as an underrepresented ethnic minority in science. One or more of the authors of this paper self-identifies as a member of the LGBTQ+ community. One or more of the authors of this paper received support from a program designed to increase minority representation in science. While citing references scientifically relevant for this work, we also actively worked to promote gender balance in our reference list. The author list of this paper includes contributors from the location where the research was conducted who participated in the data collection, design, analysis, and/or interpretation of the work.Attached Files
Published - 1-s2.0-S2211124721002989-main.pdf
Submitted - 2020.10.14.340026v1.full.pdf
Supplemental Material - 1-s2.0-S2211124721002989-mmc1.pdf
Files
2020.10.14.340026v1.full.pdf
Additional details
- PMCID
- PMC8133395
- Eprint ID
- 106092
- Resolver ID
- CaltechAUTHORS:20201015-152733216
- U19 AI109762
- NIH
- U19 AI142785
- NIH
- Scripps Research Institute
- DE-AC02-76SF00515
- Department of Energy (DOE)
- P41GM103393
- NIH
- Created
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2020-10-16Created from EPrint's datestamp field
- Updated
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2023-07-18Created from EPrint's last_modified field