Lentiviral Transduction of Target Cells

Target cells stably expressing GPI-m36.4 or GPI-FluIgG03 (scFv) were generated as described previously^{38 (link),43 (link)}. Briefly, 5 × 10⁴ TZM-bl, 293FT, or CEMss-CCR5 cells/well were seeded in a 24-well plate and incubated overnight. A total of 2 × 10⁶ TU of recombinant lentiviruses was added to the cells and supplemented with 8 μg/ml polybrene (Sigma). After incubation for 24 h, the transduced cells were extensively washed and cultured in complete DMEM. TZM-bl and 293FT cells expressing the transgenes were sorted and collected using a mouse anti-His tag antibody (Invitrogen Life Technologies) and phycoerythrin (PE)-conjugated goat anti-mouse IgG antibody (eBioscience), and the CEMss-CCR5 cells expressing the transgenes were sorted by GFP expression.