Nonhuman Primate IFITM Proteins Are Potent Inhibitors of HIV and SIV

Wilkins, Jordan; Zheng, Yi-Min; Yu, Jingyou; Liang, Chen; Liu, Shan-Lu

For cell-to-cell infection, we followed a system previously described in [29] . Briefly, Jurkat-inGLuc cells, expressing IFITMs or not expressing IFITMs, were spinoculated at 1,850 X g with infectious virus for 1 hour at 4ËšC followed by incubation at 37ËšC for 4 hours. Cells were then treated with 10 nM phorbol myristate acetate (PMA) for 4 hours. The Jurkat-inGLuc cells were then washed and refed for 18 hours at 37ËšC. The infected Jurkat-inGLuc donor cells were split in half for either cell-free or cell-to-cell infection. For cell-free infection, Jurkat-inGLuc cells were incubated for 48 hours at 37ËšC. Newly generated virus was used to infect SupT1 target cells for 48 hours. Media from SupT1 target cells was assayed for guassia Luciferase activity. For cell-to-cell infection, Jurkat-inGLuc cells were incubated with an equivalent number of SupT1 cells expressing or not expressing IFITMs. At 48 hours post-coculture, supernatants were assayed for guassia Luciferase activity using a Perkin Elmer plate reader.