The grant numbers covering the experiments are 06/1137 and 11/0342. All animal experiments were performed under S2-conditions in the central animal facility of Hannover Medical School. NOD/Ltj (NOD) and NOD.CB17-Prkdcscid/J (NOD scid) mice were bred in specific pathogen-free facilities at Hannover Medical School. For hydrodynamic gene transfer experiments 15-20 μg of the plasmid were diluted in a total volume of 2 mL phosphate-buffered saline (PBS) and injected rapidly intravenously into NOD mice. The unpaired Student 2-tailed t test or one-way analysis of variance (ANOVA) with Tukey’s posttest was performed using the GraphPad Prism program: *difference significant with P  ≤  0.05;

**very significant, P  ≤  0.01; ***extremely significant, P  ≤  0.001; P  >  0.05 was considered to be not significant (ns). Additional methods are described in the Supporting Material. Selleckchem CHIR 99021 The new induced emAIH model described here is based on an immunological danger signal combined with expression of a hepatic autoantigen. We used self-limited, hepatotropic adenovirus as a danger signal to trigger the immune response and the human and murine FTCD as common autoantigen in AIH. To this end, FTCD (Supporting Fig. 1A) and eGFP as control were cloned in a replication-deficient adenoviral vector.

Cell lines were eGFP-positive under the fluorescence microscope after transducing with Ad-eGFP (data not shown). Western blots were stained with human anti-LC1-positive serum that recognized SCH727965 in vitro human FTCD. The human anti-LC-1 serum bound to hFTCD-transduced cells, but not to control infected cells (Supporting Fig. 1B). Therefore, FTCD was expressed in hepatocytes after Ad-FTCD infection in a tertiary structure which could recognize by human autoimmune sera. As described by others,[11] adenoviral infection per

se can be used to induce a transient hepatic inflammation in animal models. Therefore, NOD mice were infected with 1 × 10[10] particles of Ad-hFTCD and Ad-eGFP. The levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were elevated at week 1 and week 2 in both groups. Values reached up to 900 and 1200 U/L of serum for AST and ALT, respectively, but returned to serum levels around the upper normal limit at later timepoints Reverse transcriptase (Fig. 1A). This ALT and AST elevation was transient and not significantly different between mice infected with Ad-hFTCD or Ad-eGFP, which was in line with all hepatitis-like animal models up to date.[12-14] Ad-eGFP transduced hepatic cells, therefore eGFP-expressing cells were detectable in the liver transiently at days 1 and 5, but not at day 20 (Fig. 1B). Likewise, adenoviral DNA could not be identified by polymerase chain reaction (PCR) from liver tissue at week 12 after infection, demonstrating the self-limiting course of acute adenoviral infection (Supporting Fig. 2).