Figure 7.

Acute mTOR inhibition abrogates inflammation induced priming.

(a) Splenocytes were cultured for 14 h in the presence of IL-15 at 100 ng/mL with or without mTOR inhibitors as indicated. Cells were then stained and analyzed by flow cytometry. The bar graphs show the averaged MFI (+/− s.d.) for pS6 and GzmB in gated NK cells (n=5 independent experiments with 1 mouse in each). (b) Rapamycin treated or non-treated mice were injected with poly(I:C) and sacrificed 18h later. Splenocytes were analyzed by flow cytometry. The graphs show the averaged MFI for pS6 and GzmB in gated NK cells. Each dot represents a single mouse, bars indicate average and SD (n=9 mice in 4 independent experiments for poly(I:C) and 18 mice in 4 independent experiments for Rapa+poly(I:C)). The MFI was normalized to control non-poly(I:C) injected mice. (c) In vivo cytotoxicity toward missing-self targets was assessed as described in the Experimental Procedures section. The percentage of remaining cells was calculated and is shown. Each dot represents a single mouse, bars indicate average and SD (n=9 mice in 4 independent experiments for poly(I:C) and 18 mice in 4 independent experiments for Rapa+poly(I:C)). (d) Human PBMCs were cultured for 36h as indicated. Cells were then stained and analyzed by flow cytometry. The bar graphs show the averaged MFI for pS6 and GzmB in gated NK cells (n=9 individual donors in 3 independent experiments, paired t-test, *p<0.05, **p<0.01).