FIG. 2.

Optimizing the VSV-G-pseudotyped retroviral infection on primary PBMCs. A GFP/VSV-G retrovirus was used to determine the optimal conditions for infecting PBMCs. (A) Optimal MOI. Following isolation, PBMCs were stimulated with PHA and rIL-2 for 2 days and then infected with GFP/VSV-G retroviruses at different MOIs. Expression of GFP was analyzed on day 4 after retroviral infection with a FACScan. (B) Expression period. PBMCs were infected with GFP/VSV-G retroviruses at an MOI of 80, and GFP expression was measured at different time points after retroviral infection. (C) Infection frequency. The cells were infected with GFP/VSV-G retroviruses at an MOI of 80 for 1, 2, 3, or 4 consecutive days starting from day 0, and GFP expression was measured on day 4. The white peaks are the PBMCs infected with S2/VSV-G control retroviruses; the black peaks are the PBMCs infected with GFP/VSV-G retroviruses.