The Study Of The Mechanism IMB-LA Inhibits Vpu Mediated Down-regulation BST-2

Host restriction factors are capable of inhibiting the replication of a series of viruses. BST-2(Tetherin/CD317) has been recently recognized as a potent interferon-induced antiviral molecule that inhibits the release of diverse mammalian enveloped virus particles from infected cells. However, several viruses have evolved countermeasures that inactivate BST-2, with the prototype being the HIV-1Vpu protein. Blocking Vpu-mediated down-regulation of cell surface BST-2provides clearly an opportunity for developing anti-HIV-1drugs.In this study, we applied epMotion5070into Cell-ELISA assay. The criteria or parameters for evaluating the suitability of epMotion5070in an HTS assay are proved to meet the requirement. The results demonstrated that epMotion5070in Cell-ELISA assay is sensitive, specific and can be used to a high-throughput screening for novel anti-HIV-1compounds.We screened50,000compounds using Cell-ELISA assay dependent on epMotion5070. Finally, we identified3leading compounds IMB-AZ, IMB-LA, IMB-DLA, which can restore BST-2surface expression by one fold in the present of Vpu. We chose one of leading compound IMB-LA to further study. In IMB-LA treated cells BST-2accumulated in CD63+late endosomes. This suggested that IMB-LA can specifically inhibit BST-2transfer from late endosomes to lysosomes, then inhibit the degradation of BST-2. HIV-1replication in cell lines expressing BST-2is effectively inhibited by the treatment of IMB-LA, this result suggests IMB-LA inhibits HIV-1replication in a BST-2dependent manner.