LuSIV cells: A reporter cell line for the detection and quantitation of a single cycle of HIV and SIV replication

A single cycle of viral replication is the time required for a virus to ent er the host cell, replicate its genome, and produce infectious progeny viri ons. The primate lentiviruses, human immunodeficiency virus (HIV) and simia n immunodeficiency virus (SIV), require on average 24 h to complete one cyc le of replication. We have now developed and characterized a reporter assay system in CEMx174 cells for the quantitative measurement of HIV/SIV infect ion within a single replication cycle. The SIV(mac)239 LTR (-225 --> +149) was cloned upstream of the firefly luciferase reporter gene and this report er plasmid is maintained in CEMx174 cells under stable selection. This cell line, designated LuSIV, is highly sensitive to infection by primary and la boratory strains of HIV/SIV, resulting in Tat-mediated expression of lucife rase, which correlates with viral infectivity. Furthermore, manipulation of LuSIV cells for the detection of luciferase activity is easy to perform an d requires a minimal amount of time as compared to current HIV/SIV detectio n systems. The LuSIV system is a powerful tool for the analysis of HIV/SIV infection that provides a unique assay system that can detect Virus replica tion prior to 24 h and does not require virus to spread from cell to cell. Thus these cells can be used for the study of replication-deficient viruses and the high throughput screening of antivirals, or other inhibitors of in fection. (C) 2000 Academic Press.