Fig. 3: Administration of rCD5L reduces lethality of WT mice subjected to high-grade CLP.

a WT mice underwent CLP to induce severe disease. Mice were then injected with two doses of 2.5 mg/kg of rCD5L intraperitoneally (IP, left) or intravenously (IV, right) 3 and 6 h post-surgery. Kaplan–Meier curves compared survival between treated and untreated groups, with significance determined by log-rank (Mantel-Cox) test. Pooled data from 4 independent experiments, number of mice per group (n) is indicated. b Protocol for analysis after rCD5L treatment. Top - analysis at 6 h. Mice were injected IP or IV with 2.5 mg/kg rCD5L at 3 h post-CLP, or PBS, and euthanized 3 h later. Bottom - analysis at 24 h. Mice were injected IP or IV with 2 doses of rCD5L, 3 and 6 h after CLP, or PBS, and euthanized at 24 h. Image created with BioRender.com. c Flow cytometry-based absolute counts of cellular populations in the peritoneum of mice injected via IP or IV routes. Leukocytes, CD45⁺; neutrophils, CD45⁺CD11b⁺Ly6G⁺; macrophages, CD45⁺CD11b⁺CD11c^-F4/80⁺; B1 cells, CD45⁺CD19⁺CD5⁺; B2 cells, CD45⁺CD19⁺CD5^-; T cells, CD45⁺CD3⁺. Statistical comparisons were made using two-tailed unpaired t-tests with Welch’s correction. Mice per group: for IP-treatment groups, for Leukocytes, Neutrophils, and Macrophages at 6 h: 6 untreated and 5 IP-treated; at 24 h: 7 untreated and 6 IP-treated. For B1, B2, and T lymphocytes at 6 h: 3 untreated and 3 IP-treated; at 24 h: 3 untreated and 2 IP-treated. In all groups of IV-treatment, there were 6 mice each. d Fold change in indicated cytokines between IP- or IV-treated mice and PBS-injected controls. Cytokines quantified by bead-based multiplex immunoassays on samples from peritoneum and serum. e CFU counts in tissues from IP- or IV-treated mice and untreated groups. Statistical differences between groups were analyzed by two-tailed Mann-Whitney test. DL, detection limit = 20 CFU. Mice per group: for IP-treatment groups, at 6 h: 6 untreated and 6 IP-treated; at 24 h: 7 untreated and 6 IP-treated. For IV-treatment groups, 6 mice were in each group. Pooled data from at least 2 independent experiments (c–e). Floating bars show the minimum, average, and maximum values within each group.