Anti-HIV Assay and Cytotoxicity Assays.
Anti-HIV and cytotoxicity assays were performed in parallel using the green fluorescent protein (GFP)-based HOG.R5 reporter cell line that was constructed and developed specifically for quantitating HIV-1 infectivity.
The system was validated and adapted as a moderately high-throughput procedure for screening natural products for anti-HIV activity in our laboratory.
Briefly, a reporter cell line for quantitating HIV-1 replication was developed using HOS
(human osteosarcoma) cells rendered susceptible to HIV-1 infection by the transfection of genes for CD4 and CCR5, the co-receptor utilized by macrophage-tropic (R5) HIV-1 isolates.
This microtiter assay is based on the transactivation of a stably integrated HIV-1 LTR-GFP transcription unit.