Objective: In this study, we investigated the effects of low intensity pulsed ultrasound (LIPUS) on the proliferation and anti-lung-tumor effect of human peripheral blood lymphocytes (HPBL). Methods: Separation of HPBLs by Ficoll gradient centrifugation. The proliferation of HPBL was stimulated by the intensity of LIUPS between 0 and 80mW/cm_². The proliferation activity of the cells was measured by the CCK-8 assay. The expressions of anti-tumor factors (Interferon-γ (INF- γ), interleukin 2 (IL-2), tumor necrosis factor α (TNF- α) and granulocyte-macrophage colony stimulating factor (GM-CSF) in the supernatant of culture medium were detected by the ELISA assay. The antitumor activity was detected by co-culture of HPBLs and human lung giant cell carcinoma cells (95D). Results: Our study shows that, compared to the control group (0 mW/cm_² ), the proliferation of HPBL cells was enhanced (P < 0.01) after 2 days of stimulation. The intensity of LIPUS is 30 mW/cm_² and duration is 10 minutes a day. With anti-tumor factor, the stimulation group’s INF- γ(39.66ng/L±8.30), IL-2 (619.84 pg/mL ± 125.94), TNF- α(461.29 ng/L± 14.08) and GM-CSF (108.11 ng/L± 2.07).They were higher than the control group (24.40ng/L±6.84), (279.56 pg/mL ± 128.18), (417.17 ng/ ± 21.44) and (96.53 ng/L± 1.56), respectively; Co-culture experiment showed that the killing rate of tumor was 51.11%±5.21 in the 30 mW/cm_² group and 42.86%±0.55 in the control group. Conclusion: The proliferation activity and killing rate of HPBLs were increased after LIPUS stimulation. The results may have great therapeutic potential and basis for the use of LIPUS combined with lymphocytes in the treatment of tumor.