Endodomain truncation of the HIV-1 envelope protein improves the packaging efficiency of pseudoviruses

Hui Zhang; Tingting Deng; Qianjiao Fang; Shaoyong Li; Shuangquan Gao; Wenling Jiang; Gege Chen; Kunyu Yu; Lizhi Zhou; Tingting Li; Qingbing Zheng; Hai Yu; Shaowei Li; Ningshao Xia; Ying Gu

doi:10.1016/j.virol.2022.07.003

Endodomain truncation of the HIV-1 envelope protein improves the packaging efficiency of pseudoviruses

Virology. 2022 Sep:574:1-8. doi: 10.1016/j.virol.2022.07.003. Epub 2022 Jul 12.

Authors

Hui Zhang¹, Tingting Deng¹, Qianjiao Fang¹, Shaoyong Li², Shuangquan Gao², Wenling Jiang¹, Gege Chen¹, Kunyu Yu¹, Lizhi Zhou¹, Tingting Li¹, Qingbing Zheng¹, Hai Yu¹, Shaowei Li¹, Ningshao Xia³, Ying Gu⁴

Affiliations

¹ State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, School of Public Health, Xiamen University, Xiamen, Fujian, 361102, China; National Institute of Diagnostics and Vaccine Development in Infectious Diseases, Xiamen University, Xiamen, Fujian, 361102, China.
² National Institute of Diagnostics and Vaccine Development in Infectious Diseases, Xiamen University, Xiamen, Fujian, 361102, China.
³ State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, School of Public Health, Xiamen University, Xiamen, Fujian, 361102, China; National Institute of Diagnostics and Vaccine Development in Infectious Diseases, Xiamen University, Xiamen, Fujian, 361102, China; The Research Unit of Frontier Technology of Structural Vaccinology of Chinese Academy of Medical Sciences, Xiamen, Fujian, 361102, China.
⁴ State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, School of Public Health, Xiamen University, Xiamen, Fujian, 361102, China; National Institute of Diagnostics and Vaccine Development in Infectious Diseases, Xiamen University, Xiamen, Fujian, 361102, China. Electronic address: guying@xmu.edu.cn.

PMID: 35858511
DOI: 10.1016/j.virol.2022.07.003

Abstract

HIV-1 remains one of the most devastating infectious pathogens without available vaccines. A valid neutralization assay using multiple representative virus strains is prerequisite for antibody response analysis in HIV-1 vaccine development, where HIV pseudoviruses (PsVs) commonly serve as surrogate agents for the authentic HIV, offering a safer manipulation in Biosafety Level 2+. However, PsV production is of low efficiency and is unstable in this field. Here, we optimize PsV production conditions via the use of alternative host cells, packaging ratios and gene truncation. We show that a 153-aa truncation of the endodomain substantially enhances the packaging efficiency of HIV PsVs, providing 4 to 25 times higher infection titers than the full-length Env. Further, we obtained a robust HIV-1 PsV panel covering 12 representative global strains for neutralization assay testing. This work sheds light on how to optimize HIV PsV packaging, and provides functional insight into the cytoplasmic domain of HIV-1.

Keywords: Endodomain truncation; HIV-1; Neutralization assay; Pseudovirus; Virus packaging.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Antibodies, Neutralizing
HIV Antibodies
HIV Infections*
HIV-1* / genetics
Humans
Neutralization Tests

Substances

Antibodies, Neutralizing
HIV Antibodies