In vivo MRI and ex vivo quantification of iron and Kupffer cells demonstrate residual phagocytic activity in mouse liver after a gadolinium chloride injection

Abstract

Kupffer cells (KCs), the resident macrophages of the liver, display a phagocytic activity that is not well quantified in animal models. Its experimental invalidation in rodents has been carried out by various means, among which the gadolinium chloride (GdCl₃) injection has been widely used, and has been generally monitored by ex vivo techniques. The aim of our study was to determine the KC phagocytic activity induced in mouse liver following a single GdCl₃ injection, through Magnetic Resonance Imaging (MRI) measurement of liver uptake of Ferumoxide in vivo, and through ex vivo quantification of Perls positive and F4/80 labeled macrophages. In this study, we showed that 24 h after an IV injection at a dose of 50 mg/kg body weight, GdCl₃ did not induce any hepato-cellular damage, nor did it strongly suppress liver phagocytic activity, as demonstrated by the persistent hepatic uptake of the iron-based MRI contrast agent Ferumoxide. In the GdCl₃-treated mice, the injection of Ferumoxide produced an increase in the liver proton transverse relaxation rate R2 which averaged 71 ± 24% of that of the control animals. The ex vivo iron and immune phenotypic quantification, performed after the Ferumoxide injection and MRI, confirmed the presence of activated phagocytes in the liver of the GdCl₃-treated animals, with a global iron score and F4/80 positive cell count respectively averaging 85 ± 26% and 46 ± 13% of their values in the untreated mice. In vivo MRI evaluation of the liver phagocytic activity using Ferumoxide may further prove useful in the follow up of both experimental and human pathologies.