Rapamycin inhibits KGF-induced mortality, proliferation, cytokine expression, and susceptibility to IAV infection, in vivo. Rapamycin inhibits KGF-induced.

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Rapamycin inhibits KGF-induced mortality, proliferation, cytokine expression, and susceptibility to IAV infection, in vivo. Rapamycin inhibits KGF-induced mortality, proliferation, cytokine expression, and susceptibility to IAV infection, in vivo. Mice were pretreated with i.p. rapamycin for 4 d before i.t. administration of PBS alone or PBS containing KGF (5 mg/kg) with and without IAV. (A) Twenty-four hours after inoculation, immunohistochemical staining for phospho-S6 was performed on lung sections and scored using a semiquantitative scale in a blinded manner. (B) Forty-eight hours after challenge with KGF, lungs were harvested and dissociated, and dispersed lung cells were stained with antibodies to pro SP-C, phospho-S6, and Ki67 and examined by flow cytometry to determine the percentage of pro SP-C + Ki67-positive cells. (C) Seventy-two hours after challenge with KGF, lungs were harvested and dissociated, and dispersed lung cells were stained with antibody to pro SP-C and IAV-NP and examined by flow cytometry to determine the percentage of pro SP-C + IAV-NP positive cells. (D) Vital status was monitored over the course of 9 d after IAV infection of rapamycin-pretreated mice and controls (P < 0.05 for KGF/IAV vs. KGF/IAV/rapa). (E) IL-6 levels were measured by ELISA in whole-lung homogenates at 48 h after infection. (F) Western blotting was performed on whole-lung homogenates isolated 24 h after IAV infection. Blots were stained with antibodies to IAV-NP and to actin as a loading control. ***P < 0.001; **P < 0.01; *P < 0.05. Nikolaos M. Nikolaidis et al. PNAS 2017;114:32:E6613-E6622 ©2017 by National Academy of Sciences