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.　©　2018,　Science　Press.　All　right　reserved.