Background: Immune function strongly influences the outcome of patients with non-small cell lung cancer(NSCLC). It’s vital to understand the immune state of patients through detecting the level of lymphocyte subsets including percentage and number timely and accurately, and helpful to evaluate conditions of prognosis and adjust the treatment program for patients.Methods: Blood sample was obtained from 172 patients with NSCLC and 55 normal controls(NCs).The absolute count and percentages of CD3+,CD3+CD4+,CD3+CD8+, B and NK cells were determined by flow cytometry simultaneously with single platform technologies. Then 172 patients were followed up for the study of effects of the lymphocyte on disease progression and disease free survival(DFS). The factors affecting disease progression were analyzed by Binary Logistic regression.Calculating survivals were with the method of Kaplan-Meier. The log-rank test and proportional hazard regression model (enter method) were used for univariable and multivariable analyses respectively. Results: The absolute counts of CD3+, CD3+CD4+, CD3+CD8+, B and NK cells were significantly decreased in patients contrasted with NCs(P<0.05), no difference in percentages(P>0.05) except NK cells between patients and NCs. Relative to NCs, patients with NSCLC at stage Ⅰ and Ⅱ had lower absolute count of lymphocyte subsets(P<0.01), patients at stage Ⅲ~Ⅳ were also significantly lower compared with those in control group (P<0.01), no significant differences were observed in the percentages between patients and NCs. Compared the absolute counts of CD3+, CD3+CD4+, CD3+CD8+, B and NK cells among patients at different stages, only CD3+CD4+ was associated with clinical stages obviously. Patients with treatment of chemotherapy after surgery or only surgery showed different effects on absolute counts of CD3+CD4+,CD3+CD8+, NK and B cells . Binary Logistic regression showed CD3+CD4+ and CD3+CD8+ could contribute to favorable prognosis. Multivariate analysis of prognostic factors of DFS showed CD3+CD4+ cell was independent factor for predicting DFS .Conclusions:Compared to NCs, patients with NSCLC at different stages showed decreased absolute lymphocyte count obviously, rather than lymphocyte percentage, which persists for months or years after chemotherapy. The high level of baseline absolute CD3+CD4+ cells count(>502 cells/ul) contributed to longer disease free survival.Chinese Clinic Trial Registry number:ChiCTR-IOR-17014139; Registry date:2017/12/25