Objective To establish the model for allergic contact dermatitis (ACD) both in vitro and in vivo. Methods The mouse model of ACD was induced by DNFB, which was verified by calculating ear swelling and HE staining. Then, the T cells isolated from ACD mouse were detected to analyze the proliferation and CD4 expression. Furthermore, ELISA was used to detect the contents of IL-2 and IL-4. Results The mouse model of ACD was established successfully, accompanied by the significant increase in ear swelling and inflammatory cell infiltration. Meanwhile, the T lymphocytes in ACD mice showed more higher responsivity and CD4 expression in vitro than in the normal control. The peak of IL-2 was much higher than that of IL-4. Conclusions The method of establishing the model of ACD both in vivo and in vitro can be used to evaluate the risk of diseases for contacting allergen and to screen some novel anti-inflammatory agents and immunomodulators.