an inducible cell-cell fusion system with integrated ability to measure the efficiency and specificity of hiv-1 entry inhibitors诱导信息融合系统与综合能力来衡量的效率和特异性hiv - 1进入抑制剂.pdfVIP

An Inducible Cell-Cell Fusion System with Integrated Ability to Measure the Efficiency and Specificity of HIV-1 Entry Inhibitors 1 1 2 2 2 Alon Herschhorn , Andres Finzi , David M. Jones , Joel R. Courter , Akihiro Sugawara , Amos B. Smith III2, Joseph G. Sodroski1,3* 1 Department of Immunology Cancer and AIDS, Dana-Farber Cancer Institute and Department of Microbiology and Immunobiology, Harvard Medical School, Boston, Massachusetts, United States of America, 2 Department of Chemistry, University of Pennsylvania, Philadelphia, Pennsylvania, United States of America, 3 Department of Immunology and Infectious Diseases, Harvard School of Public Health, Boston, Massachusetts, United States of America Abstract HIV-1 envelope glycoproteins (Envs) mediate virus entry by fusing the viral and target cell membranes, a multi-step process that represents an attractive target for inhibition. Entry inhibitors with broad-range activity against diverse isolates of HIV-1 may be extremely useful as lead compounds for the development of therapies or prophylactic microbicides. To facilitate the identification of such inhibitors, we have constructed a cell-cell fusion system capable of simultaneously monitoring inhibition efficiency and specificity. In this system, effector cells stably express a tetracycline-controlled transactivator (tTA) that enables tightly inducible expression of both HIV-1 Env and the Renilla luciferase (R-Luc) reporter protein. Target cells express the HIV-1 receptors, CD4 and CCR5, and carry the firefly luciferase (F-Luc) reporter gene under the control of a tTA- responsive promoter. Thus, Env-mediated fusion of these two cell types allows the tTA to diffuse to the target cell and activate the expression of the F-Luc protein. The eff