FIG. 7.
Robust HIV-1 protein expression, efficient Gag processing, and release of high levels of infectious HIV-1 from provirally transfected primary rabbit T cells. T cells from humans (left panels) or rabbits (right panels) were transfected with pHIV-1NL4-3 and, in parallel, with the proviral reporter pHIV-1NL4-3 GFP (transfection efficiency). After 36 h, supernatants and cells were harvested. (A) Western blots of cell lysates from three independent donors probed consecutively with anti-p24CA, anti-HIV-1 Nef, and anti-Vpu. MAPK expression served as a loading control. φ, untransfected control. Identification (ID) numbers indicate different donors. (B) Transfection efficiency determined by the percentage of viable, GFP-positive cells. (C, D) Concentrated, sucrose-pelleted virions were analyzed for p24CA concentration (C), and the infectious titer is depicted as the relative infectivity per ng p24CA (D). (B to D) Individual results from experiments with T cells from four human and eight rabbit donors (filled symbols) and the arithmetic means ± SEM of results (open symbols) are depicted. (E) Microscopic images of TZM-bl cells infected with HIV-1 virions derived from rabbit T cells and stained with a β-galactosidase substrate.