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Title: Structure-Based Design of a Soluble Prefusion-Closed HIV-1 Env Trimer with Reduced CD4 Affinity and Improved Immunogenicity

Abstract

The HIV-1 envelope (Env) trimer is a target for vaccine design as well as a conformational machine that facilitates virus entry by transitioning between prefusion-closed, CD4-bound, and coreceptor-bound conformations by transitioning into a postfusion state. Vaccine designers have sought to restrict the conformation of the HIV-1 Env trimer to its prefusion-closed state as this state is recognized by most broadly neutralizing, but not nonneutralizing, antibodies. We previously identified a disulfide bond, I201C-A433C (DS), which stabilizes Env in the vaccine-desired prefusion-closed state. When placed into the context of BG505 SOSIP.664, a soluble Env trimer mimic developed by Sanders, Moore, and colleagues, the engineered DS-SOSIP trimer showed reduced conformational triggering by CD4. Here, we further stabilize DS-SOSIP through a combination of structure-based design and 96-well-based expression and antigenic assessment. From 103 designs, we identified one, named DS-SOSIP.4mut, with four additional mutations at the interface of potentially mobile domains of the prefusion-closed structure. We also determined the crystal structures of DS-SOSIP.4mut at 4.1-Å resolution and of an additional DS-SOSIP.6mut variant at 4.3-Å resolution, and these confirmed the formation of engineered disulfide bonds. Notably, DS-SOSIP.4mut elicited a higher ratio of tier 2 autologous titers versus tier 1 V3-sensitive titers than BG505 SOSIP.664. DS-SOSIP.4mut alsomore » showed reduced recognition of CD4 and increased thermostability. We conclude the improved antigenicity, thermostability, and immunogenicity of DS-SOSIP.4mut suggest utility as an immunogen or a serologic probe; moreover, the specific four alterations identified here, M154, M300, M302, and L320 (4mut), can also be transferred to other HIV-1 Env trimers of interest to improve their properties.« less

Authors:
 [1];  [1];  [1];  [1];  [1];  [1];  [1];  [1];  [2];  [1];  [1];  [1];  [1];  [1];  [1];  [1]; ORCiD logo [1];  [1];  [1];  [1]
  1. National Inst. of Health, Bethesda, MD (United States). National Institute of Allergy and Infectious Diseases (NIAID)
  2. Frederick National Lab. for Cancer Research, Frederick, MD (United States)
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
Sponsoring Org.:
National Institute of Allergy and Infectious Diseases (NIAID); National Institutes of Health (NIH); International AIDS Vaccine Initiative (IAVI) Neutralizing Antibody Consortium; Frederick National Laboratory for Cancer Research; Leidos Biomedical Research, Inc.; USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22)
OSTI Identifier:
1357627
Grant/Contract Number:  
HHSN261200800001E; W-31-109-Eng-38
Resource Type:
Journal Article: Accepted Manuscript
Journal Name:
Journal of Virology
Additional Journal Information:
Journal Volume: 91; Journal Issue: 10; Journal ID: ISSN 0022-538X
Publisher:
American Society for Microbiology
Country of Publication:
United States
Language:
ENGLISH
Subject:
59 BASIC BIOLOGICAL SCIENCES; HIV-1; immunogen design; protein stabilization

Citation Formats

Chuang, Gwo-Yu, Geng, Hui, Pancera, Marie, Xu, Kai, Cheng, Cheng, Acharya, Priyamvada, Chambers, Michael, Druz, Aliaksandr, Tsybovsky, Yaroslav, Wanninger, Timothy G., Yang, Yongping, Doria-Rose, Nicole A., Georgiev, Ivelin S., Gorman, Jason, Joyce, M. Gordon, O'Dell, Sijy, Zhou, Tongqing, McDermott, Adrian B., Mascola, John R., and Kwong, Peter D. Structure-Based Design of a Soluble Prefusion-Closed HIV-1 Env Trimer with Reduced CD4 Affinity and Improved Immunogenicity. United States: N. p., 2017. Web. doi:10.1128/JVI.02268-16.
Chuang, Gwo-Yu, Geng, Hui, Pancera, Marie, Xu, Kai, Cheng, Cheng, Acharya, Priyamvada, Chambers, Michael, Druz, Aliaksandr, Tsybovsky, Yaroslav, Wanninger, Timothy G., Yang, Yongping, Doria-Rose, Nicole A., Georgiev, Ivelin S., Gorman, Jason, Joyce, M. Gordon, O'Dell, Sijy, Zhou, Tongqing, McDermott, Adrian B., Mascola, John R., & Kwong, Peter D. Structure-Based Design of a Soluble Prefusion-Closed HIV-1 Env Trimer with Reduced CD4 Affinity and Improved Immunogenicity. United States. doi:10.1128/JVI.02268-16.
Chuang, Gwo-Yu, Geng, Hui, Pancera, Marie, Xu, Kai, Cheng, Cheng, Acharya, Priyamvada, Chambers, Michael, Druz, Aliaksandr, Tsybovsky, Yaroslav, Wanninger, Timothy G., Yang, Yongping, Doria-Rose, Nicole A., Georgiev, Ivelin S., Gorman, Jason, Joyce, M. Gordon, O'Dell, Sijy, Zhou, Tongqing, McDermott, Adrian B., Mascola, John R., and Kwong, Peter D. Wed . "Structure-Based Design of a Soluble Prefusion-Closed HIV-1 Env Trimer with Reduced CD4 Affinity and Improved Immunogenicity". United States. doi:10.1128/JVI.02268-16. https://www.osti.gov/servlets/purl/1357627.
@article{osti_1357627,
title = {Structure-Based Design of a Soluble Prefusion-Closed HIV-1 Env Trimer with Reduced CD4 Affinity and Improved Immunogenicity},
author = {Chuang, Gwo-Yu and Geng, Hui and Pancera, Marie and Xu, Kai and Cheng, Cheng and Acharya, Priyamvada and Chambers, Michael and Druz, Aliaksandr and Tsybovsky, Yaroslav and Wanninger, Timothy G. and Yang, Yongping and Doria-Rose, Nicole A. and Georgiev, Ivelin S. and Gorman, Jason and Joyce, M. Gordon and O'Dell, Sijy and Zhou, Tongqing and McDermott, Adrian B. and Mascola, John R. and Kwong, Peter D.},
abstractNote = {The HIV-1 envelope (Env) trimer is a target for vaccine design as well as a conformational machine that facilitates virus entry by transitioning between prefusion-closed, CD4-bound, and coreceptor-bound conformations by transitioning into a postfusion state. Vaccine designers have sought to restrict the conformation of the HIV-1 Env trimer to its prefusion-closed state as this state is recognized by most broadly neutralizing, but not nonneutralizing, antibodies. We previously identified a disulfide bond, I201C-A433C (DS), which stabilizes Env in the vaccine-desired prefusion-closed state. When placed into the context of BG505 SOSIP.664, a soluble Env trimer mimic developed by Sanders, Moore, and colleagues, the engineered DS-SOSIP trimer showed reduced conformational triggering by CD4. Here, we further stabilize DS-SOSIP through a combination of structure-based design and 96-well-based expression and antigenic assessment. From 103 designs, we identified one, named DS-SOSIP.4mut, with four additional mutations at the interface of potentially mobile domains of the prefusion-closed structure. We also determined the crystal structures of DS-SOSIP.4mut at 4.1-Å resolution and of an additional DS-SOSIP.6mut variant at 4.3-Å resolution, and these confirmed the formation of engineered disulfide bonds. Notably, DS-SOSIP.4mut elicited a higher ratio of tier 2 autologous titers versus tier 1 V3-sensitive titers than BG505 SOSIP.664. DS-SOSIP.4mut also showed reduced recognition of CD4 and increased thermostability. We conclude the improved antigenicity, thermostability, and immunogenicity of DS-SOSIP.4mut suggest utility as an immunogen or a serologic probe; moreover, the specific four alterations identified here, M154, M300, M302, and L320 (4mut), can also be transferred to other HIV-1 Env trimers of interest to improve their properties.},
doi = {10.1128/JVI.02268-16},
journal = {Journal of Virology},
issn = {0022-538X},
number = 10,
volume = 91,
place = {United States},
year = {2017},
month = {3}
}

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