AnxA2 C9A mutation exacerbates host immunity in polymicrobial sepsis mice.

(A) 293T cells and neutrophils from anxa2^-/- mice were transfected with AnxA2 WT, C9A, C133A, C223A, C262A, and C335A plasmids, respectively. After H₂O₂ treatment (5 mM, 30 min), ROS levels were measured using H₂DCF assay. (B) anxa2^-/- mice were tail vein injected with these plasmids, respectively. 24 h later, mice were sacrificed and AnxA2 mRNA relative abundance in different organs or tissues were measured. (C) After plasmids transfection successfully, the mice were subjected to polymicrobial sepsis, and ROS levels were detected in PMs from mice at 24 h post-CLP. Means+SD from triplicate. (D, E) Immunostaining was performed for Ly6G⁺IL-17A⁺ cell accumulation detection in colon tissues from mice above. Fluorescence scores were quantified. (F) Peritoneal and blood bacteria were determined using CFU assay, means±SD from 3 mice. (G) Survival of mice (n = 6) above with polymicrobial sepsis was monitored. Kaplan-Meier survival curves (p = 0.0016, Log-rank Test). A-C, means+SD from 3 mice. D, E, data are representative from three independent experiments. One-way ANOVA (Tukey’s post hoc); *, p<0.05; **, p<0.01. Scale bar = 5 μm.