Fig. 8: CD5L reduces endotoxin and DAMP levels, and enhances mouse survival in septic shock.
From: CD5L as a promising biological therapeutic for treating sepsis
a ELISA quantification of LPS in sera from WT and CD5L− mice 72 h after mid-grade CLP; and in sera from WT mice subjected to high-grade CLP followed by rCD5L treatment. In these, mice were IP- or IV-injected with PBS (untreated), or with two doses of 2.5 mg/kg rCD5L, 3 and 6 h after surgery. Sera was collected and analyzed at 24 h. DL, detection limit: 0.16 ng/ml. The experimental groups comprised 7 WT and 6 CD5L− mice for mid-grade CLP, and for high-grade CLP: 7 IP-untreated, 5 IP-treated, 8 IV-untreated, and 6 IV-treated mice. b, c HMGB1 protein expression in WT and CD5L− organs, determined by immunofluorescence. b Representative images of HMGB1 expression in paraffin-preserved sections of lung, liver, and kidney of WT and CD5L− mice. c Quantification of HMGB1-stained areas in 3 regions from 2 independent images, from 2 mice per CLP group (72 h), or from 1 naïve control mouse per group (0 h), normalized by the DAPI-stained area in the corresponding regions. d, e HMGB1 expression in organs of WT mice subjected to high-grade CLP, and IP- or IV-treated with two doses of rCD5L. d Representative images of HMGB1 expression in lung, liver, and kidney of treated and untreated mice. e Quantification of HMGB1-stained areas, as in (c). a, c, e Floating bars represent the minimum, average and maximum values within each group. Statistical differences between groups were analyzed using a two-tailed Mann-Whitney test. b, d Scale bar: 20 μm. f WT and CD5L− mice were IP-injected with a sublethal LPS dose (1.5 mg/kg). Survival was monitored for 6 days. g WT mice were injected with a lethal LPS dose (10 mg/kg), and 3 h later with 2.5 or 5 mg/kg rCD5L, or left untreated (0). Survival was monitored for 4 days. f, g Kaplan–Meier curves were generated to compare survival between groups. Significance was determined by log-rank (Mantel-Cox) test. The graphical representation includes pooled data from 3 independent experiments for (f) and 2 independent experiments for g.
