| Objective:To establish DC+CIK therapy standard;To compare therapeutic efficacy of DC+CIK plus chemotherapy and chemotherapy alone,and to evaluate clinical effective power of DC+CIK therapy in lung cancer comprehensively and objectively.Methods:1.PBMCs obtained from NSCLC patients,and add various kind of cytokines to prepare DC and CIK.DCs were added into CIK cultures on the seventh day.After the coculturing for seven days,the killing activity of CIK cells were detected by LDH,and the cytokines secreted by CIK cells were detected by ELISA/ ELISPOT.Then the phenotypes of CIK cells were detected by flow cytometry;2.In clinical study,DC+CIK and chemotherapy were administered at intervals.The survival rate of 120 NSCLC patients,including 50 post-op patients and 70 advanced patients,were analysed,The clinical therapeutic effect were observed via imaging and biochemistry recce,and the quality of life were evaluated according to KPS;3. Disease free survival rate and overall survival rate were compared between biotherapy plus chemotherapy groups and conventional chemotherapy groups;4.Ten NSCLC patients were divided into two groups according to clinical therapeutic effects.The cytokines in serum of the patients before and after biotherapy were detected by human cytokine antibody array,and the relationship of the clinical therapeutic effect and the change of cytokine was evaluated.T cell subsets and NK cells in peripheral blood of lung cancer patients before and after biotherapy were detected by flow cytometry.Results:1.LDH essay showed that the cytotoxicity of DC+CIK on tumor cells was higher than that of CIK alone(P＜0.05).ELISA essay showed that the levels of TGF-βand IL-10 in DC+CIK were significantly lower than those in CIK alone (P＜0.05),and the level of IFN-γwas significantly higher in DC+CIK(P＜0.05).The level of Tregs in DC+CIK group were significantly lower than those of CIK alone (P＜0.05).2.No serious adverse effects were observed after transfusion except low-grade fever observed in some patients.The 2 years disease free survival rate and overall survival rate of post-op patients received biotherapy were 77.8±6.3%and 93.2±3.9%,respectively.The 2 years disease free survival and overall survival rate in patients who received biotherapy more than 4 times were higher than those received biotherapy 4 times or less respectively(82.9±7.9%vs 56.0±12.1%,P＜0.05;100%vs 83.5±8.9%).3.For advanced patients who received biotherapy,their 1,2 and 3 years survival rate were 59.6±5.9%,29.6±6.1%,21.8±6.0%,respectively.The 2 years survival rate in patients who received biotherapy 3 times or more were higher than those received biotherapy less than 3 times(47.1±9.8%vs 15.4±6.5%,P＜0.05).4. The improvement of quality of life were more obviously in post-op patients than in advanced patients after biotherapy(77.8%vs 68%),however,the difference did not achieve a statistical significance;5.In post-op patients,the 2 years overall survival rate of those recerved biotherapy plus chemotherapy was higher than those received conventional chemotherapy(94.7±3.6%vs 78.8±7.0%,P＜0.05).The 2 years survival rate of advanced patients who received biotherapy plus chemotherapy was higher than those received conventional therapy(27.0±6.3%vs 10.1±4.6%,P＜0.05).6.The serum analysis of 10 NSCLC patients who received biotherapy showed that the levels ofIL-2,IL-5,IL-6,MCP-2,and MCP-3 increased after biotherapy,while the levels of other cytokine,such as TGF-β,IL-1α,IL-8,and MCP-1,decreased or unchanged. Patients who were benefited from biotherapy showed an increase in the levels of IFN-γ,G-CSF,GRO,GRO-a,IL-10,IL-13,MIG,TNF-α,and TNF-βin serum when compared with patients developed recurrence/metastasis after biotherapy.Conclusion:In basic study,we found that the cytotoxicity of the CIK cells on tumor cells was elevated after coculturing with DC.In clinical research,we have confirmed that the applying of DC+CIK therapy can prolong survival time,raise the quality of life and improve immune condition of NSCLC patients.
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