αgalcer

Male 8–10 week old C57BL/6 mice (Jackson Labs, Bar Harbor, Maine) were used. All procedures were approved by the University of Calgary Animal Care Committee (protocol numbers AC14-0129, AC14-0128) and were performed in accordance with the guidelines of the Canadian Council on Animal Care. The Con A hepatitis model is a widely used and well-characterized model of immune-mediated liver injury that mimics many aspects of AIH in patients (19 (link), 20 (link)). Mice were treated with Con A (13.5 mg/kg iv; Sigma, St. Louis, MO) or vehicle (PBS) (26 (link)), and were sacrificed 16 h later. Hepatitis severity was determined biochemically (ALT) and histologically (H & E staining) (26 (link)). A second mouse model of immune-mediated liver injury, resulting from the administration of alpha galactosylceramide (αGalCer), was used to assess the impact of mirtazapine treatment on hepatitis severity. In this well-characterized model a single intraperitoneal injection of αGalCer (100 μg/kg; Cayman Chemical, Ann Arbor, Michigan, USA) induces a moderate hepatitis (maximal at 16 h post-αGalCer administration) that is independent of hepatic macrophage/monocyte activation (27 (link), 28 (link)). All experiments were repeated at least twice unless otherwise noted.

At the week 4 after infection, mice were treated intravenously (i.v.) with 10 μg of the NKT cell agonist α-GalCer (KRN7000, Cayman Chemical Company, USA) and euthanized four weeks later to determine the fungal burden in the lung.