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

被引:77
作者
Chuang, Gwo-Yu [1 ]
Geng, Hui [1 ]
Pancera, Marie [1 ]
Xu, Kai [1 ]
Cheng, Cheng [1 ]
Acharya, Priyamvada [1 ]
Chambers, Michael [1 ]
Druz, Aliaksandr [1 ]
Tsybovsky, Yaroslav [2 ]
Wanninger, Timothy G. [1 ]
Yang, Yongping [1 ]
Doria-Rose, Nicole A. [1 ]
Georgiev, Ivelin S. [1 ]
Gorman, Jason [1 ]
Joyce, M. Gordon [1 ]
O'Dell, Sijy [1 ]
Zhou, Tongqing [1 ]
McDermott, Adrian B. [1 ]
Mascola, John R. [1 ]
Kwong, Peter D. [1 ]
机构
[1] NIAID, Vaccine Res Ctr, NIH, 9000 Rockville Pike, Bethesda, MD 20892 USA
[2] Leidos Biomed Res Inc, Frederick Natl Lab Canc Res, Canc Res Technol Program, Electron Microscopy Lab, Frederick, MD USA
基金
美国国家卫生研究院;
关键词
HIV-1; immunogen design; protein stabilization; IMMUNODEFICIENCY-VIRUS TYPE-1; ENVELOPE GLYCOPROTEIN COMPLEX; NEUTRALIZING ANTIBODY; CRYSTAL-STRUCTURE; MONOCLONAL-ANTIBODIES; VACCINE DESIGN; GP140; TRIMERS; GP120; EPITOPES; MODEL;
D O I
10.1128/JVI.02268-16
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
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-wellbased 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-angstrom resolution and of an additional DS-SOSIP. 6mut variant at 4.3-angstrom 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. 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. IMPORTANCE One approach to elicit broadly neutralizing antibodies against HIV-1 is to stabilize the structurally flexible HIV-1 envelope (Env) trimer in a conformation that displays predominantly broadly neutralizing epitopes and few to no nonneutralizing epitopes. The prefusion-closed conformation of HIV-1 Env has been identified as one such preferred conformation, and a current leading vaccine candidate is the BG505 DS-SOSIP variant, comprising two disulfides and an Ile-to-Pro mutation of Env from strain BG505. Here, we introduced additional mutations to further stabilize BG505 DS-SOSIP in the vaccine-preferred prefusion-closed conformation. In guinea pigs, our best mutant, DS-SOSIP. 4mut, elicited a significantly higher ratio of autologous versus V3-directed neutralizing antibody responses than the SOSIP-stabilized form. We also observed an improvement in thermostability and a reduction in CD4 affinity. With improved antigenicity, stability, and immunogenicity, DS-SOSIP. 4mut-stabilized trimers may have utility as HIV-1 immunogens or in other antigen-specific contexts, such as with B-cell probes.
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页数:18
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