Figure 7
T-cells from hCD4/hCCR5/hCycT1-triple-tg rats do not release virions into the culture supernatant following infection by several R5 HIV-1 strains, despite enhanced transcriptional activity. Activated T-cells derived from the spleens of four double- or four triple-tg rats, or from PBMC of two human donors were plated in 96-well round bottom plates and were challenged with (A) single-round VSV-G HIV-1NL4-3 GFP reporter virus and on day 3 p.i., the MFI (GFP) was quantified by flow cytometry as a surrogate for early HIV-1 gene expression. (B) In parallel, T-cells were challenged with replication-competent R5 HIV-1 strains YU-2, Ba-L, or JR-FL (each 5 ng p24 per 2 × 106 cells) overnight. Cultures were washed twice with PBS and continuously cultivated for 2 weeks. At day 7 and 13 p.i., supernatant aliquots were removed and analyzed for the p24 concentration by antigen ELISA. Histograms depict the arithmetic mean ± SD of triplicates of values on day 13 p.i. from one experiment.