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It is reported that successive electroacupuncture stimulation (EAS) at different frequencies demonstrates different effects, but this feature on immunological function is largely unclear.The aim of the present study was to explore the effects of 2 Hz, 100 Hz, and 2/100 Hz EAS on immunological function in cyclophosphamide (CP)-treated mice.The results showed: (1) It was observed a reversion of the decrease in splenic natural killer cell activity induced by CP at 1 h following single 2 Hz, 100 Hz, and 2/100 Hz EAS, especially 2/100 Hz EAS.(2) 1h after single EAS, 2 Hz and 100 Hz groups but not 2/100 Hz group showed a decrease in splenic T lymphocyte proliferation CP-treated mice.(3) Compared with CP group, 100 Hz and 2/100 Hz rather than 2 Hz group presented elevations in splenic NK cell activity and splenic index, but a decline in proliferation of splenic cell after three times successive EAS.(4) The level of IL-6 in serum in all EAS groups was higher than CP-treated group after three times successive EAS.These findings suggested that low and high frequency EAS both improved the immunological function after administrating CP, and the effect of 2/100 Hz EAS was better, which may be closely related to its actions in promoting the release and synthesis ofIL-6.