A simple screening system for anti-HIV drugs:: Syncytium formation assay using T-cell line tropic and macrophage tropic HIV env expressing cell lines -: Establishment and validation

被引:29
作者
Chiba, H
Asanuma, S
Okamoto, M
Inokoshi, J
Tanaka, H [1 ]
Fujita, K
Ömura, S
机构
[1] Kitasato Univ, Sch Pharmaceut Sci, Minato Ku, Tokyo 1088641, Japan
[2] Showa Univ, Sch Med, Shinagawa Ku, Tokyo 1140064, Japan
[3] Kitasato Univ, Kitasato Inst Life Sci, Minato Ku, Tokyo 1088641, Japan
关键词
D O I
10.7164/antibiotics.54.818
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
The first step in cellular entry of HIV involves binding of the viral envelope glycoprotein complex (gp120/gp41) to specific receptor molecules on the target cells. The cell-cell fusion (syncytium formation) between env expressing cells and CD4+ cells mimics the viral infection of the host cells. To search for anti-HIV substances preventing this process, we constructed the recombinant cell lines, HeLa/CD4/Lac-Z -and HeLa/T-env/Tat for T-cell tropic (HIV-1(NL4-3)) system, and HOS/CD4/CCR5/Lac-Z and HeLa/M-env/Tat for macrophage tropic (HIV-1(SF162)) system. When each pair of cells were co-incubated for 20 hours, the multinuclear giant cells (syncytia) were formed and beta -galactosidase was expressed. These systems are less biohazardous because no infectious virus particles are used. Their validity in screening for anti-HIV substances which inhibit syncytium formation was confirmed using various known HIV entry inhibitors.
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收藏
页码:818 / 826
页数:9
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