a screen against leishmania intracellular amastigotes comparison to a promastigote screen and identification of a host cell-specific hit屏幕对利什曼虫细胞内无鞭毛体相比promastigote屏幕和宿主特异性的识别.pdfVIP

A Screen against Leishmania Intracellular Amastigotes: Comparison to a Promastigote Screen and Identification of a Host Cell-Specific Hit 1 2 2 2 1 Geraldine De Muylder *, Kenny K. H. Ang , Steven Chen , Michelle R. Arkin , Juan C. Engel , James H. McKerrow1 1 Department of Pathology, Sandler Center for Drug Discovery, University of California San Francisco, San Francisco, California, United States of America, 2 Small Molecule Discovery Center, University of California San Francisco, San Francisco, California, United States of America Abstract The ability to screen compounds in a high-throughput manner is essential in the process of small molecule drug discovery. Critical to the success of screening strategies is the proper design of the assay, often implying a compromise between ease/ speed and a biologically relevant setting. Leishmaniasis is a major neglected disease with limited therapeutic options. In order to streamline efforts for the design of productive drug screens against Leishmania, we compared the efficiency of two screening methods, one targeting the free living and easily cultured promastigote (insect–infective) stage, the other targeting the clinically relevant but more difficult to culture intra-macrophage amastigote (mammal-infective) stage. Screening of a 909-member library of bioactive compounds against Leishmania donovani revealed 59 hits in the promastigote primary screen and 27 in the intracellular amastigote screen, with 26 hits shared by both screens. This suggested that screening against the promastigote stage, although more suitable for automation, fails to identify all active compounds and leads to numerous false positive