2 chain, hypersecrete T helper 1 (Th1) and Th2 cytokines and chemokines upon stimulation with an appropriate ligand, such as alpha-galactosylceramide (α-GalCer). In doing so, these iNKT cells exert considerable and promiscuous immune function, including altering immune regulation by activating dendritic cells (DCs), macrophages, NK cells, T cells, B cells, and driving the development of adaptive immunity.12, 13 iNKT cells appear to play a critical role in the regulation of several other autoimmune diseases, including type 1 diabetes, multiple sclerosis, rheumatoid arthritis, and systemic lupus erythematosus.13-17 Previously, our laboratory has proposed
that activation of iNKT cells is a critical factor in accelerating disease.18-20 To explore this issue buy RAD001 in depth, we immunized C57BL/6 mice with 2-OA-BSA (bovine serum albumin) and activated their iNKT cells with
α-GalCer. We report herein that iNKT cell activation by α-GalCer leads to a profound exacerbation of portal disease in 2-OA-BSA-immunized mice, including increased AMA production, increased CD8+ T cell biliary infiltration, portal inflammation, granuloma formation, bile duct damage, and fibrosis. These results are critical and emphasize the role of innate immunity in the natural history of PBC and they further suggest mechanisms by which biliary disease becomes perpetuated in humans as well as explaining the recurrence of Olaparib nmr PBC following liver transplantation in the absence of major histocompatability complex (MHC) compatibility. These data also emphasize the appearance of fibrosis, a feature thus far lacking in the other murine models of autoimmune cholangitis. 2-OA, 2-octynoic acid; α-GalCer, alpha-galactosylceramide; α-SMA, alpha-smooth muscle actin; AMAs, antimitochondrial antibodies; BSA, bovine serum albumin; CFA, complete Freund’s adjuvant; DC, dendritic cells; dnTGF-βRII, TGF-β receptor II dominant-negative; HSCs, hepatic stellate cells; iNKT, invariant natural killer T; PBC, primary biliary cirrhosis; click here PDC-E2, E2 subunits of the pyruvate dehydrogenase complex; TGF-β,
transforming growth factor beta. The protocol for induction of autoimmune cholangitis is similar to our previous reports.7 Briefly, female C57BL/6 mice aged 8-10 weeks were obtained from the National Laboratory Animal Center and maintained in the Animal Center of the College of Medicine, National Taiwan University. Mice were intraperitoneally immunized with 2-OA-BSA (100 μg) in the presence of complete Freund’s adjuvant (CFA, Sigma-Aldrich, St. Louis, MO) and subsequently boosted at weeks 2, 4, 6, 8, and 10 with 2-OA-BSA and incomplete Freund’s adjuvant (IFA, Sigma-Aldrich). Two μg of α-GalCer (Alexis, San Diego, CA) was diluted in phosphate-buffered saline (PBS) and injected intravenously 1 day before first, second, and third 2-OA-BSA immunizations (group name: α-GC/CFA/2-OA).