Inhibition of Human Immunodeficiency Virus Type 1 Entry by a Keggin Polyoxometalate

Wang, Xiaoli; Wang, Jiao; Zhang, Wenmei; Li, Boye; Zhu, Ying; Hu, Qin; Yang, Yishu; Zhang, Xiaoguang; Yan, Hong; Zeng, Yi

The anti-HIV-1 activity of PT-1 was performed using the TZM-bl assay [31] . The HIV-1 Env-pseudotyped viruses were prepared by co-transfection HEK-293T cells with HIV-1 pREJO4541.67 Env expression plasmid and pSG3∆env using FuGENE transfection reagent (Promega, Madison, WI, USA). The recombinant viruses were collected from cell supernatant after centrifugation at 2500 rpm for 5 min and stored in liquid nitrogen. The titers of HIV-1 Env-pseudotyped viruses were examined by the 50% tissue culture infective dose (TCID 50 ) assay using Reed-Muench method. Late, TZM-bl cells were seeded in the 96-well plate and infected with 1000 TCID 50 /mL virus diluted in diethylaminoethanol (DEAE) dextran (Sigma, Shanghai, China) in the presence of PT-1. Two days after infection, the infectivity of viruses was evaluated using the Bright-Glo luciferase assay kit (Promega) under the manufacturer's instructions. The luciferase activity was analyzed using the Enspire ® Multi-mode Plate Reader (PerkinElmer) and expressed in terms of relative luciferase units (RLUs). The half maximal inhibitory concentration (IC 50 ) was calculated as a 50% inhibitory concentration using GraphPad Prism 6 software.