The ability of thrombin to enhance the intrusion of HIV-1 pristine detached strain into target cells

Objective To observe the effects and reasons of thrombin on HIV envelope V3 area crown sequence in different strain by using cell-cell fusion experimental system and expend the range of strain.Methods In order to prepare theEnv-293T cell by transfecting 293T cells,plasmid pSVⅢ-ADA,plasmid pSM-06057c3 and plasmid pSM-03009c34 are selected.And these three kinds of plasmids must carry HIV-1 ADA,06057c3 and 03009c34 env genes.239T cell are transfected by plasmid pSVⅢ-ADA and pcTat,pSM-06057c3 and pcTat pSM-03009c34 and pcTat.The ratio of plamid and pcTat is 2∶ 1.After that there is a cell fusion experiment between the Env-293T cell and Magic5A cell in different Th concentrations and count the fusion cells and observe the effect of cell fusion.Results The treated cell can promote ADA and 06057c3 Env mediated fusion,whose V3 crown sequences are GPGRAF.Th can not promote 03009c34 Env mediated fusion,whose V3 crown sequence is GPGQAW strain.Conclusion Th can promote HIV meditated cell fusion,and the reason which might be related with the V3 crown GPGRAF sequences. Key words: HIV-1 ;  Thrombin ;  Pristine detached strain