SOS and IP Modifications Predominantly Affect the Yield but Not Other Properties of SOSIP.664 HIV-1 Env Glycoprotein Trimers.
SOS and IP Modifications Predominantly Affect the Yield but Not Other Properties of SOSIP.664 HIV-1 Env Glycoprotein Trimers.
复制标题
SOS 和 IP 修饰主要影响 SOSIP.664 HIV-1 Env 糖蛋白三聚体的产量，但不影响其他特性。
DOI：
10.1128/jvi.01521-19
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发表时间：
2019
影响因子：
5.4
通讯作者：
Moore,JohnP
中科院分区：
文献类型：
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作者：
Ringe,RajeshP;Colin,Philippe;Torres,JonathanL;Yasmeen,Anila;Lee,Wen-Hsin;Cupo,Albert;Ward,AndrewB;Klasse,PJ;Moore,JohnP
正在开发各种人类免疫缺陷病毒1型（HIV-1）基因型的可溶性重组天然样（NL）包膜糖蛋白（Env）三聚体，作为旨在诱导广泛中和抗体（bNAb）的候选疫苗。原型设计（命名为BG 505 SOSIP.664）包含一个亚基间二硫键（SOS），以共价连接gp 120和gp 41胞外域（gp 41 ECTO）亚基，并包含一个点取代I559 P（IP），以进一步稳定gp 41 ECTO组分。在没有SOS和IP变化的情况下，蛋白水解切割的三聚体倾向于分解成它们的组成gp 120和gp 41 ECTO亚基。然而，我们发现，缺乏SOS和/或IP变化的NL三聚体可以被亲和纯化，其量足以分析其抗原性和热稳定性。一般而言，这些三聚体变体具有与完全稳定的SOSIP.664版本高度相当的性质。我们的结论是，SOS和IP变化的主要影响是大幅增加表达过程期间和之后的三聚体稳定性，从而允许产生有用的量。然而，一旦三聚体被纯化，SOS和IP的变化对bNAb和其他表位的热稳定性和抗原性只有微妙的影响。重要的是，基于HIV-1 env基因的三聚体蛋白正在开发用于人类疫苗试验。这些蛋白质的一个特征是它们模仿中和抗体靶向的病毒颗粒上的包膜糖蛋白结构，即，防止细胞被感染的抗体。一个正在探索的疫苗概念是重组三聚体在作为免疫原递送时可能能够引发病毒中和抗体。一种常用的设计被命名为SOSIP.664，这是一个反映序列变化的术语，用于稳定三聚体并允许它们以实际有用的量生产。在这里，我们表明这些稳定化变化可以增加生产细胞内生物合成过程中的三聚体产量，但对纯化三聚体的性质影响很小。
Soluble recombinant native-like (NL) envelope glycoprotein (Env) trimers of various human immunodeficiency virus type 1 (HIV-1) genotypes are being developed as vaccine candidates aimed at the induction of broadly neutralizing antibodies (bNAbs). The prototypic design, designated BG505 SOSIP.664, incorporates an intersubunit disulfide bond (SOS) to covalently link the gp120 and gp41 ectodomain (gp41ECTO) subunits and a point substitution, I559P (IP), to further stabilize the gp41ECTOcomponents. Without the SOS and IP changes, proteolytically cleaved trimers tend to disintegrate into their constituent gp120 and gp41ECTOsubunits. We show, however, that NL trimers lacking the SOS and/or IP change can be affinity purified in amounts sufficient for analyses of their antigenicity and thermal stability. In general, these trimer variants have properties highly comparable to those of the fully stabilized SOSIP.664 version. We conclude that the major effect of the SOS and IP changes is to substantially increase trimer stability during and after the expression process, thereby allowing useful amounts to be produced. However, once the trimers have been purified, the SOS and IP changes have only subtle impacts on thermostability and the antigenicity of bNAb and other epitopes.IMPORTANCERecombinant trimeric proteins based on HIV-1envgenes are being developed for vaccine trials in humans. A feature of these proteins is their mimicry of the envelope glycoprotein structure on virus particles that is targeted by neutralizing antibodies, i.e., antibodies that prevent cells from becoming infected. One vaccine concept under exploration is that recombinant trimers may be able to elicit virus-neutralizing antibodies when delivered as immunogens. A commonly used design is designated SOSIP.664, a term reflecting the sequence changes that are used to stabilize the trimers and allow their production in practically useful amounts. Here, we show that these stabilizing changes act to increase trimer yield during the biosynthesis process within the producer cell but have little impact on the properties of purified trimers.