Objective To establish and compare the reference range of normal values of five types of lymphocyte subsets in mice of three strains by flow cytometry. Method The five types of Th1/Th2 cytokines in Balb/C, C3H/HeJ and C3H/SW mice were determined by flow cytometry. The results were statistically analyzed with SPSS 10.0 software. Resoults The distribution of the five lymphocyte subsets presents a Gaussian distribution in the 3 mouse strains, respectively. The 95% confidence interval for mean values can be accepted. The difference at lymphocyte subsets level is not significant in Balb/C and C3H/SW mice except B cells. The values of lymphocyte subsets of C3H/HeJ mice were lowest among the three strains of mice. Conclusion The normal reference values of Th1/Th2 cytokine in Balb/C, C3H/HeJ and C3H/SW mice have been established. The cellular immune function of C3H/HeJ is most feeble among the three strains, and C3H/HeJ mice may be susceptible to infection with pathogenic microorganisms.