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Design and characterization of a germ-line targeting soluble, native-like, trimeric HIV-1 Env lacking key glycans from the V1V2-loop.
Biochimica et Biophysica Acta (BBA) - General Subjects ( IF 3 ) Pub Date : 2020-09-16 , DOI: 10.1016/j.bbagen.2020.129733
Shubbir Ahmed 1 , Tripti Shrivastava 1 , Rajesh Kumar 1 , Mohit Kumar 2 , Manidipa Banerjee 2 , Naresh Kumar 1 , Manish Bansal 1 , Supratik Das 1 , Sweety Samal 1
Affiliation  

Background

The HIV-1 envelope glycoprotein (Env) is the primary target for broadly neutralizing antibodies (bNAbs) which can block infection. The current design strategy of soluble forms of Env in native-like trimeric conformation induces neutralizing antibodies with minimal breadth and potency. Extensive shielding by N-glycans on the surface of the HIV-1 Env acts as an immune evasion mechanism by restricting B cell recognition of conserved neutralizing determinants. An alternate approach is to design Env protein with glycan deletion to expose the protein surface.

Methods

A stable native-like trimeric Env with glycan holes at potentially immunogenic locations is expected to elicit better induction of germ-line B-cells due to exposure of the immunogenic regions. However, the extent and consequences of glycan removal from the trimer apex that form an important epitope is not explored. In this work, we have designed a construct with glycans deleted from the trimer apex of an Indian clade C origin Env that has previously been characterized for immunogenicity, to understand the impact of deglycosylation on the structural and functional integrity as well as on the antibody binding properties.

Results

The V1V2 glycan-deleted protein maintains native-like trimeric conformation with improved accessibility of the V1V2-directed germ-line antibodies. Furthermore, we showed that the protein binds specifically to quaternary conformation-dependent bnAbs but minimally to non-neutralizing antibodies.

Conclusions

This study provide an important design aspect of HIV-1 Env-based immunogens with glycan holes in the apex region that could be useful in eliciting apex directed antibodies in immunization studies.



中文翻译:

针对可溶的,类似天然的三聚体HIV-1 Env的种系的设计和表征,该蛋白缺乏来自V1V2环的关键聚糖。

背景

HIV-1包膜糖蛋白(Env)是广泛中和抗体(bNAb)的主要目标,该抗体可以阻止感染。当前的天然形式三聚体构象的Env可溶形式的设计策略以最小的广度和效力诱导中和抗体。通过限制保守的中和决定簇的B细胞识别,HIV-1 Env表面上N聚糖的广泛屏蔽起到了逃避免疫的作用。另一种方法是设计带有聚糖缺失的Env蛋白,以暴露蛋白表面。

方法

由于暴露于免疫原性区域,预期在潜在的免疫原性位置具有聚糖孔的稳定的天然样三聚体Env会引发更好的种系B细胞诱导。然而,尚未探索从形成重要表位的三聚体顶部去除聚糖的程度和后果。在这项工作中,我们设计了从印度进化枝C起源Env的三聚体顶点缺失聚糖的构建体,该构建体以前具有免疫原性特征,以了解去糖基化对结构和功能完整性以及对抗体结合的影响属性。

结果

V1V2聚糖缺失的蛋白保持天然样的三聚体构象,并改善了V1V2定向的种系抗体的可及性。此外,我们表明该蛋白质特异性结合季构象依赖性bnAb,但最少结合非中和抗体。

结论

这项研究提供了一个重要的设计方面,即基于HIV-1 Env的免疫原在先端区域带有聚糖孔,可用于在免疫研究中诱导先导抗体。

更新日期:2020-09-23
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