Figure 4.

Heme-albumin administration prevents anti-CD40 antibody driven inflammatory liver disease. C57Bl/6J mice were injected with heme-albumin (70 µmol/kg, i.p.) and after 4 h with anti-CD40 antibody (20 mg/kg). The animals were sacrificed at T_16h for plasma cytokines or mRNA gene expression, at T_34h for liver enzymes, and at T_52h for liver histology. (A) Representative macroscopic photographs of the liver lobe from mice injected with anti-CD40 antibody, with or without heme-albumin. Necrosis is visible as a white spot. (B) Representative macroscopic images of the spleen of mice treated with anti-CD40 antibody, with or without heme-albumin. Displayed is the average size of the spleens (mm) ± 2 SDs. (C) Heatmap displaying the color-coded plasma concentrations of IL6, TNFα, and liver enzymes (ALT and AST) in animals treated with heme-albumin (heme) or albumin (control) and challenged with or without anti-CD40 antibody (yellow=low concentration, red=high concentration). (D) Heatmap displaying the color-coded mRNA expression levels of proinflammatory cytokines in Dynabeads-enriched F4/80⁺ liver macrophages in C57BL/6J mice that were treated with heme-albumin (heme) or albumin (control) and stimulated with or without anti-CD40 antibody. These data demonstrate that acute heme-albumin treatment mitigated macrophage-driven inflammation and suppressed anti-CD40 antibody mediated disease.