Pore-Forming Toxins Induce Macrophage Necroptosis during Acute Bacterial Pneumonia
Fig 10
Inhibition of RIP1 or MLKL decreases morbidity and mortality during S. marcescens hemorrhagic pneumonia.
BALB/c mice were infected intratracheally with Sma at high dose (5.0 x 106 CFU) or low dose (1.0 x 106 CFU). A) CFU recovered from the BALF of BALB/c mice pretreated with PBS or clodronate liposomes 24h after infection with low dose Sma. Each symbol represents an individual mouse. B) Survival of mice infected with the high dose of Sma that received intraperitoneal pre-treatment with CoQ10, necrostatin-5 (N5), or N5 along with CoQ10 (N5/C10). Mice received 100 μl of a 100 μM solution of each drug intraperitoneally from time of challenge every 4h for the first 12h post-infection. C) Percent weight change and D) airway bacterial burden of Sma infected mice (n = 4–5/cohort) (low dose) when treated with PBS or the described N5/C10 therapy. The concentration of E) total leukocytes, F) neutrophils, and G) monocytes (y-axis is log scale) in Hema-3-stained cytospins of BALF from Sma infected mice (low dose) that were received mock or N5/C10 therapy. H) Airway bacterial burden, I) number of F4/80 positive cells in 50 μl, J) IL-1β levels, and K) TNFα levels in BALF from Sma infected mice (low dose) that received treatment with PBS, GW806742X (GW80; 100 μl of 100 μM), GSK’872 (100 μl of 10 μM) or ZVAD (100 μl of 10 μM). Mann-Whitney U tests were applied for two-group comparisons, for multiple group comparisons Dunn’s multiple-comparison post-test was used: *, P ≤ 0.05, **, P ≤ 0.01, ***, P ≤ 0.001. Data are representative of ≥3 separate experiments, each with 8 biological replicates.