FIG. 1.
Optimization of viral production from acute infection of T cells. (A) Strategy for high-level production of HIV-1/Δvif from acute infection of nonpermissive cells. See text for details. (B) Analysis of the benefits of VSV-G pseudotyping, spin infection, and use of HIV-1YU-2 for optimal viral output after acute infection. Pseudotyped or nonpseudotyped preparations of the indicated viruses were generated by transient transfection of 293T cells and used to infect nonpermissive HUT78 cells, either by spin infection or passive diffusion. The infected cells were then washed extensively to remove residual input particles and incubated for ∼30 h in growth medium to allow viral production to reach a high level. The cells were then washed again and placed in fresh growth medium for ∼10 h. The quantity of soluble p24Gag produced during this period was measured by ELISA.