Cleavage-independent HIV-1 Env trimers engineered as soluble native spike mimetics for vaccine design.
Cleavage-independent HIV-1 Env trimers engineered as soluble native spike mimetics for vaccine design.
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DOI：
10.1016/j.celrep.2015.03.047
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发表时间：
2015-04-28
期刊：
影响因子：
8.8
通讯作者：
Wyatt RT
中科院分区：
文献类型：
--
作者：
Sharma SK;de Val N;Bale S;Guenaga J;Tran K;Feng Y;Dubrovskaya V;Ward AB;Wyatt RT
病毒糖蛋白通过 pH 激活或受体结合激活介导进入，并以亚稳态融合前状态存在，可以通过定向理性设计来稳定该状态。正如最近报道的，融合前状态 (SOSIP) 的构象固定的 HIV-1 包膜糖蛋白 (Env) 三聚体在相对较高的分辨率下表现出分子同质性和结构完整性。然而，SOSIP 需要完全的 Env 前体裂解，这需要内源性弗林蛋白酶过度表达。在这里，我们开发了一种替代策略，使用 Env 子结构域的柔性肽共价连接来生产可溶性、均质且不依赖于裂解的 Env 模拟物，称为天然柔性连接 (NFL) 三聚体，作为候选疫苗。这种简化的设计避免了弗林蛋白酶共表达的需要，并且在一种情况下不需要抗体亲和纯化，以加速临床前和临床应用的三聚体放大。我们已成功将 NFL 设计转化为多种 HIV-1 亚型，有望成为一种为 HIV-1 或其他病毒生产类似天然、有序的 Env 三聚体的通用方法。
Viral glycoproteins mediate entry by pH-activated or receptor-engaged activation and exist in metastable pre-fusogenic states that may be stabilized by directed rational design. As recently reported, the conformationally fixed HIV-1 envelope glycoprotein (Env) trimers in the pre-fusion state (SOSIP) display molecular homogeneity and structural integrity at relatively high levels of resolution. However, the SOSIPs necessitate full Env precursor cleavage, which requires endogenous furin over-expression. Here, we developed an alternative strategy using flexible peptide covalent linkage of Env subdomains to produce soluble, homogeneous and cleavage-independent Env mimics, called native flexibly linked (NFL) trimers, as vaccine candidates. This simplified design avoids the need for furin co-expression and, in one case, antibody affinity purification to accelerate trimer scale-up for preclinical and clinical applications. We have successfully translated the NFL design to multiple HIV-1 subtypes, establishing the potential to become a general method of producing native-like, well-ordered Env trimers for HIV-1 or other viruses.