Autoimmune hepatitis (AIH) is characterized by an immune‐mediated injury of the hepatic parenchyma of unknown pathogenesis. Type 2 AIH is identified by the presence of anti‐liver‐kidney microsomes type 1 (anti‐LKM1) and anti‐liver cytosol type 1 (anti‐LC1) autoantibodies. The current study shows that a murine model of AIH can be generated by DNA immunization against type 2 AIH self‐antigens (P450 2D6 and formiminotransferase‐cyclodeaminase). A pCMV plasmid containing the N‐terminal region of mouse CTLA‐4 and the antigenic region of human CYP2D6 (672‐1,377 bp) and human formiminotransferase cyclodeaminase (FTCD; 1,232‐1,668 bp) was used for DNA immunization of C57BL/6 female mice. Immunized mice showed elevated levels of alanine aminotransferase (ALT), with peaks at 4 and 7 months postinjection. Periportal, portal, and intralobular liver inflammatory infiltrates were observed at histology. Mainly CD4+ lymphocytes, but also CD8+ and B lymphocytes, were found in the liver. Cytotoxic‐specific T cells were found in both the liver and spleen of these animals. Mice developed anti‐LKM1 and anti‐LC1 antibodies of immunoglobulin G2 (IgG2) subclass, against specific mouse autoantigens. The ALT levels correlated with both the presence of anti‐LKM1/anti‐LC1 antibodies and the presence of liver necroinflammation. In conclusion, in mice, DNA immunization against human autoantigens breaks tolerance and induces an autoimmune liver disease. Molecular mimicry between foreign and self‐antigens explains the liver injury. This model of AIH resembles human type 2 AIH and will be helpful for the study of its pathogenesis. (HEPATOLOGY 2004;39:1066–1074.)