PD-1单克隆抗体对体外诱导扩增的CIK细胞抗肺癌效应的影响  

Influence of PD-1 monoclonal antibody on anti-lung cancer effect of cytokine-induced killer cells(CIK)induced and expanded in vitro

作　　者：刘通 王贺双 许凝 LIU Tong;WANG Heshuang;XU Ning(Graduate School of Dalian Medical University,Dalian,116044,Liaoning,P.R.China;Department of Thoracic Surgery,Dalian Municipal Central Hospital Affiliated to Dalian Medical University,Dalian,116044,Liaoning,P.R.China;Department of Central Laboratory,Dalian Municipal Central Hospital Affiliated to Dalian Medical University,Dalian,116044,Liaoning,P.R.China)

机构地区：[1]大连医科大学研究生院,辽宁大连116044 [2]大连医科大学附属大连市中心医院胸外科,辽宁大连116044 [3]大连医科大学附属大连市中心医院中心实验室,辽宁大连116044

出　　处：《中国胸心血管外科临床杂志》2021年第8期990-997,共8页Chinese Journal of Clinical Thoracic and Cardiovascular Surgery

基　　金：大连市科技创新基金项目(2018J13SSN128)。

摘　　要：目的探讨程序性死亡受体1(programmed cell death protein-1,PD-1)单克隆抗体对体外诱导扩增的细胞因子诱导杀伤细胞(cytokine-induced killer cells,CIK)抗肺癌效应的影响。方法收集大连市中心医院肿瘤内科2019年1~5月确诊的20例中晚期肺癌患者[男8例、女12例,平均年龄42~65(56.45±5.89)岁]外周血单个核细胞,体外诱导扩增为CIK细胞,设PD-1单克隆抗体联合CIK细胞培养组、单独CIK细胞培养组和PD-1单克隆抗体组进行实验,采用CCK-8法分别检测不同效靶比(E∶T)条件下各组CIK细胞对肺癌A549细胞杀伤活性,用流式细胞仪检测PD-1单克隆抗体联合CIK细胞培养组和单独CIK细胞培养组穿孔素及颗粒酶表达阳性比例,ELISA法检测白细胞介素2(IL-2)、干扰素-γ(IFN-γ)及肿瘤坏死因子-α(TNF-α)细胞因子分泌水平。结果 CCK8法检测CIK细胞对肺癌A549细胞的杀伤效应随效靶比增大而增大,PD-1单克隆抗体提高了不同效靶比条件下CIK细胞对A549肺癌细胞的杀伤效应,E∶T为5∶1(28.5%±1.9%vs. 20.3%±1.8%)、10∶1(40.6%±2.4%vs.31.7%±2.1%)、20∶1(57.4%±3.5%vs. 44.7%±3.8%)、40∶1(74.1%±8.3%vs. 60.8%±5.3%),PD-1单克隆抗体联合CIK细胞组和单独CIK细胞组在不同效靶比条件下对肺癌A549细胞的杀伤效应差异具有统计学意义(P<0.05),两组细胞对肺癌A549细胞杀伤效应均强于单独PD-1单克隆抗体组(P<0.01)。流式细胞仪检测结果提示抗PD-1单克隆抗体提高了CIK细胞的穿孔素和颗粒酶释放阳性比例,CIK细胞联合PD-1单克隆抗体组与单独CIK细胞组穿孔素释放阳性比例(46.7%±3.5%vs. 35.1%±2.2%)和二者颗粒酶释放水平(34.6%±3.8%vs. 25.7%±3.3%)差异具有统计学意义(P<0.05)。与单独CIK细胞组相比,PD-1单克隆抗体联合CIK细胞组提高了IL-2、TNF-α、IFN-γ细胞因子的分泌水平[(5 409.0±168.8)pg/mL vs.(4 300.0±132.3)pg/mL,(252.7±16.7)pg/mL vs.(172.5±8.6)pg/mL,(327.2±23.5)pg/mL vs.(209.7±16.0)pg/mL],各组间差Objective To investigate the influence of programmed cell death protein-1(PD-1)monoclonal antibody on the anti-lung cancer effect of cytokine-induced killer cells(CIK)which were programmed in vitro.Methods Peripheral blood mononuclear cells from 20 patients(8 males and 12 females with an average age of 56.45±5.89 years ranging from 42 to 65 years)diagnosed with advanced lung cancer from January to May 2019 at the Department of Oncology of Dalian Central Hospital were collected and induced to amplify into CIK cells in vitro.PD-1 monoclonal antibody combined with CIK cell culture group,individual cell culture group and PD-1 monoclonal antibody group were set up to detect the cell killing activity of CIK cells against lung cancer under different effective target ratio conditions,and the ratio of perforin and granzyme positive expression in PD-1 monoclonal antibody combined CIK cell culture group and individual CIK cell culture group was detected by flow cytometry.ELISA method was used to detect the interleukin-2(IL-2),interferon-γ(IFN-γ)and tumor necrosis factor-α(TNF-α)cytokine secretion levels in the two groups.Results The killing effect of CIK cells on A549 lung cancer cells increased with the increase of effective target ratio by CCK8,and PD-1 monoclonal antibody increased the killing effect of CIK cells on A549 lung cancer cells under different effective target ratio,E∶T=5∶1(28.5%±1.9%vs.20.3%±1.8%),10∶1(40.6%±2.4%vs.31.7%±2.1%),20∶1(57.4%±3.5%vs.44.7%±3.8%),40∶1(74.1%±8.3%vs.60.8%±5.3%).The killing effect of PD-1 monoclonal antibody combined with CIK cells and CIK cells alone on A549 lung cancer cells was statistically different(P<0.05).The killing effect of cells in both groups on lung cancer A549 cells was stronger than that of the PD-1 monoclonal antibody group(P<0.01).The results of flow cytometry showed that PD-1 monoclonal antibody increased the positive ratio of perforin and granzyme release in CIK cells,and the positive ratios of perforin release(46.7%±3.5%%vs.35.1%±2.2%)and gr

关 键 词：PD-1单克隆抗体 细胞因子诱导杀伤细胞(CIK) 肺癌 免疫治疗 

分 类 号：R734.2[医药卫生—肿瘤]