Therapeutic Intervention with Anti-Complement Component 5 Antibody Does Not Reduce NASH but Does Attenuate Atherosclerosis and MIF Concentrations in Ldlr-/-.Leiden Mice

Abstract

Background: Chronic inflammation is an important driver in the progression of non-alcoholic steatohepatitis (NASH) and atherosclerosis. The complement system, one of the first lines of defense in innate immunity, has been implicated in both diseases. However, the potential therapeutic value of complement inhibition in the ongoing disease remains unclear. Methods: After 20 weeks of high-fat diet (HFD) feeding, obese Ldlr-/-.Leiden mice were treated twice a week with an established anti-C5 antibody (BB5.1) or vehicle control. A separate group of mice was kept on a chow diet as a healthy reference. After 12 weeks of treatment, NASH was analyzed histopathologically, and genome-wide hepatic gene expression was analyzed by next-generation sequencing and pathway analysis. Atherosclerotic lesion area and severity were quantified histopathologically in the aortic roots. Results: Anti-C5 treatment considerably reduced complement system activity in plasma and MAC deposition in the liver but did not affect NASH. Anti-C5 did, however, reduce the development of atherosclerosis, limiting the total lesion size and severity independently of an effect on plasma cholesterol but with reductions in oxidized LDL (oxLDL) and macrophage migration inhibitory factor (MIF). Conclusion: We show, for the first time, that treatment with an anti-C5 antibody in advanced stages of NASH is not sufficient to reduce the disease, while therapeutic intervention against established atherosclerosis is beneficial to limit further progression.

Ldlr-/-.Leiden mice develop neurodegeneration, age-dependent astrogliosis and obesity-induced changes in microglia immunophenotype which are partly reversed by complement component 5 neutralizing antibody

“…Until the start of the studies, the animals were fed a standardized chow diet (Sniff R/M V1530, Uden, the Netherlands). Randomization, blinding methods and power calculations were used as previously described (Seidel et al, 2022).…”

“…During the last 12 weeks of HFD feeding, one group of mice received intraperitoneal injections with an established anti-C5 antibody (BB5.1, 5 mg/mL in PBS, 1 mg/mouse; Zelek et al, 2020). The BB5.1 antibody was produced as reported (Seidel et al, 2022). A HFD-fed control group received intraperitoneal injections of PBS (as a vehicle control).…”

“…To unravel potential underlying biological processes in the brain, gene expression was analysed in the hippocampus, the most important brain region involved in memory and learning. Finally, we tested in HFD-fed Ldlr-/-.Leiden mice whether an anti-complement therapy inhibiting systemic complement C5 (BB5.1 antibody; Zelek et al, 2020), shown to improve vascular function in the same mouse model (Seidel et al, 2022), can limit neurodegeneration and neuroinflammation.…”

Differential effects of plant and animal fats on obesity‐induced dyslipidemia and atherosclerosis in Ldlr−/−.Leiden mice

“…Plasma total cholesterol and triglycerides were determined using the enzymatic assays Cholesterol Gen.2 and TRIGL respectively (both Roche Diagnostics) according to the manufacturer's instructions. Commercially available ELISA kits were downscaled 25,26 and then used according to the manufacturer's instructions for the analysis of plasma oxLDL (Cusabio), ApoB100 (Abcam), MIF, E-selectin, P-selectin, ICAM-1, and VCAM-1 (all R&D Systems).…”

Advances in Molecular and Translational Medicine

“…However, the treatment reduced the development of atherosclerosis, limiting the total lesion size and severity, independently of an effect on plasma cholesterol but with reductions in oxidized LDL (oxLDL) and the macrophage migration inhibitory factor (MIF). The study suggests that anti-C5 treatment in advanced stages of NASH is not sufficient to reduce the disease, while therapeutic intervention against established atherosclerosis could be beneficial to limit further progression [12]. The article by De Cunto et al discusses the progression of chronic obstructive pulmonary disease (COPD) and the persistence of lung inflammation even after smoking cessation.…”