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.
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.
Chiba
H
H
School of Pharmaceutical Sciences, Kitasato University, Tokyo, Japan.
Asanuma
S
S
Okamoto
M
M
Inokoshi
J
J
Tanaka
H
H
Fujita
K
K
Omura
S
S
D019380:Anti-HIV Agents / Q000494:pharmacology*; D015153:Blotting, Western; D015704:CD4 Antigens / Q000378:metabolism; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D004353:Drug Evaluation, Preclinical / Q000379:methods; D015686:Gene Products, env / Q000096:biosynthesis*; D017930:Genes, Reporter / Q000235:genetics; D015756:Genes, tat / Q000235:genetics; D015726:Giant Cells / Q000187:drug effects*; D006367:HeLa Cells; D006801:Humans; D006822:Hybrid Cells; D007763:Lac Operon / Q000235:genetics; D008264:Macrophages / Q000187:drug effects / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics; D015203:Reproducibility of Results; D013601:T-Lymphocytes / Q000187:drug effects / Q000378:metabolism*
D019380:Anti-HIV Agents; D015704:CD4 Antigens; D015686:Gene Products, env