The Preliminary Clinical Research Of Cytokine-induced Killer Cells Co-culturing With Autologous Dendritic Cells On The Clinical Immunotherapy For Lung Cancer

Background:Lung cancer is the most common kind of carcinomar in the world.The 5 year survival rate is still less than 15%nowdays.Immunotherapy has become the hot studying focous of tumor research.Since Lymphokine actirated Killer cells by IL-2 had been used in the end stage carcinoma treatment by Doctor Rosenberg,and had achieved good clinical efficacy in 1980s,there are many kinds of cells used for the immunotherapeutics of malignant tumors.Immunotherapeutical cells include T lymphocyte,natural killer cells,dendritic cells and cytokine-induced killer(CIK) ceils. Researchers had found that cytokine-induced killer(CIK) cells are inoculated with dendritic cells,DCIK cells are got,which could directly kill tumor cells,and DCIK has the strong cytotoxicity against tumor cells as T lymphocytes,and it also has the MHC-unrestrictive killing characterics of Natural killer cells(NK).And it has been identified in animals that reinfusion with DCIK into animals underlying malignant carcinomar could get broad antitumor activity,with great safety and efficance.Objects:It's to explore a novel kind of effector cell against tumor cells.It's to investigate the cytotoxicity in vitro of cytokine-induced killer(CIK) cells after co-culturing with autologous dendritic cells(DC),and to compare it with CIK's cytotoxicity.And then to observe their clinical antitumour efficacy,side effects and immunological activity.Methods:Peripheral blood mononuclear cells(PBMC) were isolated from 12 patients with lung cancer,who had been treated with standard protocols.Dendritic Cells and CIK cells had been respectively inoculated in vitro for 1 week,we observed the morphous of DC through contrast phase microscope,and then inoculated them together for 10-14 days according to the ratio 1:5.Then we determined the phenotype of Cytokine- induced Killer Cells after co-incubated with autologous DCs(DCIK cells) through through flow cytometer,and observed the morphous of DCIK cell through contrast phase microscope.We tested the cytotoxicity of DCIK against A549s in vitro on different ratios through MTT asssy,and compared the cytotoxicity of DCIK cells with that of CIK cells.All the patients had been treated with nearly 1×10~9 DCIK cells per time for immunotherapy by reinjecting into themseleves.Each patient had accepted 3 courses and each course included 3 times.And then we evaluated their immunological activity, clinical antitumour efficacy and side effects.Results:1.After cocultured for 14 days,when the effector-to-target ratios(DCIK:A549 or CIK:A549) were 1.0:1,1.25:1,2.5:1,5:1,10:1,the lysis ratios of DCIK cells were relatively 22.92±1.89%,30.91±2.13%,45±3.63%,52.83±2.29%,62.12±5.46%;the lysis ratios of CIK cells were relatively17.98±0.31%,20.82±1.51%,34.50±2.50%,47.20±2.27%,50.11±4.32%;P＜0.05.2.Before the DCIK immunotherapy,the percentage of CD3~+T lymphocyte is 75.25±9.56%;after the DCIK immunotherapy,the percentage of CD3~+T lymphocyte is 75.50±9.79%,t=0.198,P＞0.05.Before the DCIK immunotherapy,the percentage of CD3~+CD4~+ T lymphocyte is 24.44±10.89%,after the DCIK immunotherapy,the percentage of CD3~+CD4~+ T lymphocyte is 24.44±10.89%, t=3.69,P＜0.05;Before the DCIK immunotherapy,the percentage of CD3~+CD8~+ T lymphocyte is 30.69±14.05%,after the DCIK immunotherapy,the percentage of CD3~+CD8~+ T lymphocyte is41.68±15.98%,t=3.82,P＜0.05;Before the DCIK immunotherapy,the percentage of CD3~+CD56~+ T lymphocyte is 10.96±4.85%, after the DCIK immunotherapy,the percentage of CD3~+CD56~+ T lymphocyte is 24.89±4.27%,t=12.53,P＜0.05.The ratios of CD3~+CD4~+,CD3~+CD8~+,CD3~+CD56~+ significantly increased(P＜0.05),comparing with ratios before DCIK cells infusion.3.Among the 12 patients,1 patient got complete response,4 patients got partial response and 1 patient got stable disease,The other 6 patients got disease progressed,including that 2 patients died.so the recent clinical efficacy rate is 41.6%,and the rate of disease control is 50%.4.No patient got side effects other than 2 patients who took a brief sensation of heat.Conclusions:1.The cytotoxicity of DCIK cells is better than CIK's in vitro.2.In the immunotherapy for lung cancer,reinjection of DCIK cells could increase the basic immunological level,improve the anti-tumor immune response of the patients,and DCIK cell is a novel kind of effectors against tumor cells.3.DCIK cells could get better clinical efficacy,and it could be an adjunctive therapy beside the operation,the chemotherapy,the radiotherapy and other treatment strategies.4.DCIK cells have good safety.