In this study, we compared the effects of video-assisted thoracic surgery (VATS) and traditional open surgery (TOS) on immune system functioning in non-small cell lung cancer (NSCLC) patients. We enrolled 122 NSCLC patients in this study. The patients were randomly divided into VATS group (n = 61) and TOS group (n = 61). Plasma DNA concentration was analyzed by fluorescence quantitative PCR. Automatic blood analyzer was used to measure WBC-C, and immune nephelometry was employed to assess hs-CRP concentrations. The number of CD3+T, CD4+T and CD8+T lymphocytes in peripheral blood was estimated by flow cytometry. ELISA was used to quantify the levels of IGFBP-3, VEGF and IL-6. Compared to the TOS group, surgery-related blood loss and pain score on day 1 after surgery were lower in VATS group. After surgery, the out-of-bed activity occurred earlier and in-hospital stays were shorter in the VATS group compared to the TOS group. Plasma free DNA concentration of VATS group patients at first, third and fifth days after surgery was lower than that of the TOS group. WBC-C and hs-CRP levels were lower in the VATS group at each time point after surgery. The number of CD3+T, CD4+T, CD8+T lymphocytes and CD4+/CD8+ was lower in the TOS group compared to VATS group. Plasma IGFBP-3, VEGF and IL-6 levels were significantly lower in VATS group compared to the TOS group. Finally, incidence of complications in the VATS group was dramatically lower than the TOS group (all P < 0.05). Based on our findings, compared to TOS, VATS significantly decreased the incidence of acute-phase reaction and lowered the inhibition of immune functions after surgery.