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Background: This study aimed at establishing the autoimmune hepatitis (AIH) model of C57BL/6 mice, and examining the expression and significance of T follicular helper (Tfh) cells, T follicular regulatory (Tfr) cells, effector B cells and other indicators in this experimental autoimmune hepatitis (EAH) model. Methods: C57BL/6 mice in experimental group were administered by intraperitoneal injection after fully emulsified on 1st day and 7th day with 0.5 mL of 0.5–2.0 g/L S-100 and an equal volume of Buddha Complete Adjuvant (CFA). The levels of serum alanine transaminase (ALT), aspartate aminotransferase (AST), and interleukin-21 (IL-21) were tested by the corresponding kit. Tfr, Tfh and B cell subsets were quantified by flow cytometry. Histological pathology was completed by pathological section experiments. Results: In comparison with that in the healthy controls (HC), significantly increased numbers of serum ALT and AST, Tfr, IL-21 was observed. The hepatic lobules and hepatocyte cords were severely disorganized. Conclusion: The results of liver pathological changes and serum index changes were similar to the chronic and progressive pathogenesis and pathology of AIH patients, providing a detailed pathological basis for the basic research and clinical experiments of AIH.