Malaria-Induced NLRP12/NLRP3-Dependent Caspase-1 Activation Mediates Inflammation and Hypersensitivity to Bacterial Superinfection
Figure 3
NLRP3/NLRP12-dependent activation of caspase-1 and pyroptosis in mice infected with P. chabaudi.
(A) At 9 days post-infection, splenocytes from C57BL/6, and P2X7R−/− mice were lysed and analyzed by Western blot employing caspase-1-specific antibody. (B) At 7 days post-infection, active caspase-1 by FLICA reagent, membrane integrity by propidium iodide, and cell size change by shift on FSC axis were assessed in splenic macrophages (CD11b+F4/80+) and DCs (CD11c+MHC-II+). (C) Splenocytes lysates from mice at 7 days post-infection were used to detected active caspase-1 by Western blot. A faint band of similar molecular weight of active caspase-1 that corresponds to IgG light chain is seen in the uninfected controls or in various infected knockout mice. The results presented in figures 3A, 3B and 3C are representative of 2 experiments that yield similar results. (D) A LPS dose of 10 µg/mouse was given intravenously at 7 days post-infection with P.chabaudi and sera collected 9 hours later, for measuring IL-1β and TNF-α levels. The numbers within parenthesis indicate the percentage of lethality 48 hours after LPS challenge (10 µg/mouse). The levels of IL-1β measured in the sera of infected C57BL/6, NLRP3−/−, NLRP12, ASC−/− and Casp-1−/− were not different. The results are means + SEM of 10 mice from 2 independent experiments. Significant differences are indicated by **p<0.001 obtained in the Mann-Whitney test.