Tat-based cell-cell fusion method for screening HIV-1 fusion inhibitors.

Xiaoli Wang; Yishu Yang; Sisi Shen; Xianliang Wang; Tian Feng; Qin HU; Yi Zeng

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Tat-based cell-cell fusion method for screening HIV-1 fusion inhibitors.

Author: Xiaoli WANG ¹ ; Yishu YANG ¹ ; Sisi SHEN ¹ ; Xianliang WANG ² ; Tian FENG ¹ ; Qin HU ¹ ; Yi ZENG ¹
Author Information

1. College of Life Science and Bioengineering, Beijing University of Technology, Beijing 100124, China.
2. National Institute of Environmental Health Chinese Center for Disease Control and Prevention, Beijing 100123, China.
Publication Type:Journal Article
Keywords: HIV-1; cell-cell fusion; drug screening; entry inhibition
From: Chinese Journal of Biotechnology 2018;34(3):429-439
CountryChina
Language:Chinese
Abstract: An HIV-1 cell-cell fusion system was developed to screen HIV-1 entry inhibitors that block cell-cell fusion. In this system, the pEGFP-Tat plasmid was constructed and cotransfected into effector cells (HEK-293T) with HIV-1 envelope plasmid. TZM-bl cell, a genetically engineered cell line that expresses CD4, CXCR4, CCR5 as well as Tat-inducible β-galactosidase and luciferase reporter gene, was used as target cell. Thus, the co-culture of target cells and effector cells allows the cell fusion via Env and the activity of the fusion inhibitor can be quantified by measuring the reporter protein expression. The experimental parameters were optimized and 11 anti-HIV-1 agents including CCR5 antagonist maraviroc, reverse transcription inhibitor zidovudine (AZT) and integrase inhibitor raltegravir were tested. The result showed that the system exhibited high specificity and sensitivity. Two of eight tested anti-HIV-1 agents were found to block the cell-cell fusion. The system is suitable for efficient screening of HIV-1 cell-cell fusion inhibitors.