Testing of VIF Inhibitors Against Clinical Isolates and Animal Study Support

Sanders-Beer, Brigitte

Abstract

Core B will support the overall grant application by providing specific in vitro and in vivo tests of promising Vifinhibitor candidates.
The first aim i s to test the efficacy and toxicity of promising Vif inhibitor candidates inhuman PBMC-based assays using clinical HIV-1 isolates including neurotropic isolates.
The second aim i sto test promising Vif inhibitor candidates in the HIV-1 Hollow Fiber Mouse model. Hollow fibers filled withHIV-1 infected cells will be implanted subcutaneously and intraperitoneally into Balb/c mice. Subsequently,the mice will be treated intraperitoneally with the Vif inhibitor candidates. The mice will be monitored foracute toxicity and the fibers evaluated for efficacy against HIV-1 replication.
The third aim i s to performquantitative RNA viral load determinations on rhesus macaque plasma and viral load samples in support ofthe SIV/primate studies at New England Primate Research Center (Project 3). For the quantitative viral loadmeasurements, TAQMAN RT-PCR using an ABI7900 sequencer will be performed. Primers, probe, andstandard are from the gag region of SIV.The relevance of this project to public health:Currently, about 1 million people are infected with HIV in the United States. In addition, 40,000 newinfections are occurring each year. So far, about 500,000 persons died from complications of HIV-infectionin the US. In the last 15 years, a variety of anti-HIV compounds have been developed but these compoundsonly control but do not cure the infection. All of the compounds currently licensed are directed against twoHIV enzymes or against virus/cell fusion. Recently, the interaction of another HIV protein, Vif, with a cellularprotein (APOBEC3G) was discovered. The newly identified compounds that inhibit this interaction have thepotential to cure the infection with HIV-1 since Vif is required for replication in primary human cells.