Figure 4.

rhPLTP protected against inflammation and death in Pltp−/− mice with polymicrobial sepsis. (A) Repeated iv injections of rhPLTP after CLP produced a cumulative effect on plasma PLTP activity. Pltp−/− mice received iv injections of vehicle (control) or rhPLTP (25 µg in a volume of 200 µl) 2 h, 4 h, 7 h and 23 h after CLP. PLTP activity was measured in plasma before and 8 h and 24 h after CLP and expressed as a percentage of WT mouse plasma activity used as a reference (n = 8 mice per group, ANOVA with Tukey’s multiple comparisons test). (B) Repeated iv injections of rhPLTP increased survival after CLP. Pltp−/− mice received several intravenous injections of rhPLTP (25 µg in a volume of 200 µl) or vehicle at 2 h, 4 h, 7 h, 23 h, 32 h, 48 h, 56 h, 72 h, 80 h, 96 h, 104 h after CLP. Survival rates were analyzed by the Kaplan-Meier method and compared using the log rank test (n = 16–20 mice per group). (C) Repeated iv injections of rhPLTP prevented the production of inflammatory cytokines (IL-6, MCP-1, TNF-α, IL-1β) after CLP. Pltp−/− mice received several iv injections of vehicle (control) or rhPLTP (25 µg in a volume of 200 µl) 2 h, 4 h, 7 h, and 23 h after CLP. Plasma samples, harvested from Pltp−/− mice before CLP, 8 h and 24 h after CLP, were assayed using a Milliplex mouse cytokine panel (n = 15–19 mice per group, Mann-Whitney test). Data are means ± sem. *P < 0.05, **P  < 0.01, ***P < 0.001. (D) Histological examination of liver and kidney of Pltp−/− mice 24 h after CLP showed a protective effect of rhPLTP injections. The organs were collected 24 h after CLP, and fixed with paraformaldehyde prior to HE staining. The arrows indicate the presence of hepatocyte clusters with acidification. The arrowheads indicate damaged glomeruli. Representative photographs of 5 mice per group are shown (original magnification x400). Scale bars are equivalent to 200 µm.