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. 2011 Feb 17;7(2):e1001284. doi: 10.1371/journal.ppat.1001284

Figure 4. Role of cell contacts and of TLR7 on recognition of HIV-infected cells by Gen2.2 cells.

Figure 4

a. Activation of Gen2.2 requires a direct contact with HIV-infected cells. Shaking cocultures of Gen2.2 and HIV-1-infected MT4C5 (left panel), or separating infected MT4C5 cells and target Gen2.2 in a transwell chamber (right panel) impairs IFN production, whereas recognition of FLUAV by Gen2.2 is not affected. Mean+sd of 3 independent experiments is shown b. Inhibition of IFN production by Bafilomycin A1. Gen2.2 cells were cocultivated with HIV-infected MT4C5 cells, exposed to FLUAV, or to CpG (a TLR9 agonist) during 24 h, in the absence or presence of Bafilomycin A1 (Bafilo 125 nM). IFN levels are expressed as the % of the signal obtained without drugs. c. Role of TLR7. Gen2.2 cells were transduced with lentiviral vectors expressing shRNAs against TLR7 (shTLR7) or an irrelevant target (shCTRL). Levels of TLR7 mRNA in transduced cells were measured by RT-PCR. Data are normalized to GAPDH mRNA and expressed as relative levels of mRNA compared to shCTRL cells. Mean+sd of 3 independent experiments are shown (left panel). IFN production in TLR7-silenced and control Gen2.2 cells, cocultivated with HIV-infected MT4C5 or stimulated with FLUAV or CpG for 40 h. IFN levels are expressed as a percentage of the signal obtained with shCTRL Gen2.2 cells.