Document Detail

A sensitive HIV-1 envelope induced fusion assay identifies fusion enhancement of thrombin.
MedLine Citation:
PMID:  20045672     Owner:  NLM     Status:  MEDLINE    
To evaluate the interaction between HIV-1 envelope glycoprotein (Env) and target cell receptors, various cell-cell-fusion assays have been developed. In the present study, we established a novel fusion system. In this system, the expression of the sensitive reporter gene, firefly luciferase (FL) gene, in the target cells was used to evaluate cell fusion event. Simultaneously, constitutively expressed Renilla luciferase (RL) gene was used to monitor effector cell number and viability. FL gave a wider dynamic range than other known reporters and the introduction of RL made the assay accurate and reproducible. This system is especially beneficial for investigation of potential entry-influencing agents, for its power of ruling out the false inhibition or enhancement caused by the artificial cell-number variation. As a case study, we applied this fusion system to observe the effect of a serine protease, thrombin, on HIV Env-mediated cell-cell fusion and have found the fusion enhancement activity of thrombin over two R5-tropic HIV strains.
De-Chun Cheng; Guo-Cai Zhong; Ju-Xiang Su; Yan-Hong Liu; Yan Li; Jia-Ye Wang; Toshio Hattori; Hong Ling; Feng-Min Zhang
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2010-01-05
Journal Detail:
Title:  Biochemical and biophysical research communications     Volume:  391     ISSN:  1090-2104     ISO Abbreviation:  Biochem. Biophys. Res. Commun.     Publication Date:  2010 Jan 
Date Detail:
Created Date:  2010-01-27     Completed Date:  2010-03-05     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0372516     Medline TA:  Biochem Biophys Res Commun     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1780-4     Citation Subset:  IM    
Copyright Information:
Copyright 2010 Elsevier Inc. All rights reserved.
Department of Microbiology, Harbin Medical University, 194 Xuefu Road, Harbin, Heilongjiang 150081, China.
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MeSH Terms
Cell Fusion / methods
Cell Line
HIV Envelope Protein gp160 / metabolism*
HIV Fusion Inhibitors / pharmacology
HIV-1 / drug effects,  physiology*
Luciferases, Renilla / biosynthesis
Membrane Fusion*
Receptors, HIV / metabolism*
Thrombin / metabolism*
Virus Internalization*
Reg. No./Substance:
0/HIV Envelope Protein gp160; 0/HIV Fusion Inhibitors; 0/Receptors, HIV; EC, Renilla; EC

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