CD80联合CD28/CpG ODN活化的PBMC对胃癌细胞杀伤作用的实验研究  

Killing efficacy of PBMC costimulately activated by CD28/CpGODN combined with CD80 on the gastric cancerous cell:Experimental study

作　　者：方军 郭树军 李永研 

机构地区：[1]153医院普通外科,河南郑州450042

出　　处：《实用医药杂志》2010年第3期248-250,共3页Practical Journal of Medicine & Pharmacy

摘　　要：目的研究CD80和CD28/CpGODN(含CPG的核脱氧核苷酸)活化单个核细胞(PBMC)在体外对胃癌细胞株MKN45杀伤作用机理,为胃癌的过继免疫治疗可能性提供参考。方法外周血PBMC的分离与体外培养,共刺激活化外周血PBMC,用含共刺激活化PBMC或/和CD800.4mg/L的100ml/L胎牛血清的RPMI1640培养基培养细胞,测生长曲线;MTT法检测活化细胞的体外淋巴细胞毒作用,并用电镜观察效应细胞杀伤的胃癌细胞超微结构及用流式细胞仪检测结肠癌细胞的凋亡指数。结果CD80本身对胃癌细胞株MKN45有一定的杀伤作用(P<0.05),CD28/CpGODN共刺激活化PBMC在体外对胃癌细胞株MKN45杀伤作用明显(P<0.01),CD80与CD28/CpGODN共刺激活化PBMC合用,对胃癌细胞株MKN45的杀伤作用显著大于CD80本身对胃癌细胞株MKN45杀伤作用(P<0.01),亦大于单独应用CD28/CpGODN共刺激活化PBMC对胃癌细胞株MKN45的杀伤作用(P<0.05),CD80增加了胃癌细胞株MKN45对共刺激活化PBMC的敏感性。电镜结果显示,效应细胞作用24h肿瘤细胞就部分发生坏死,部分肿瘤细胞可见凋亡。流式细胞仪检测效应细胞胃癌细胞株MKN4572h明显凋亡。结论CD80与共刺激活化PBMC合用,提高了CD28/CpGODN活化PBMC对胃癌细胞株MKN45的靶向性杀伤作用。单抗协同诱导的效应细胞对胃癌细胞株MKN45杀伤效果较好,以上二者合用较单用任何一种处理对胃癌细胞株MKN45杀伤作用更强,而坏死细胞进一步增多说明效应细胞是通过诱导肿瘤细胞坏死及细胞凋亡两条途径来实现杀伤作用,从而说明活化的淋巴细胞杀伤胃癌细胞是通过坏死及凋亡实现的。Objective To investigate killing effect of PBMC costimulately activated by CD28/CpGODN by CD28 and CpGODN MoAb combined with CD80 on human gastric cancerous cell line in vitro. Method The cancer cells were cultured with PBMC and CD28/CpGODN MoAb and CD80, then the growth curve of cells was obtained. The killing efficiency was measured by MTF method. The submicroscopic structure of cells was observed by electron microscope. Apoptosis was verified by a flow eytometer. Result After treated with costimulated activation of PBMC with CD28 and CpGODN,the inhibition rate of MKN45 cells was 69.15%.When two methods were used in combination,the inhibition rate was 96.15%. By MTT method,combination of costimulated activation of PBMC with CD28/CpGODN and CD80 showed enhanced killing effect compared with single therapy （P〈0.01）. Electron microscope and flow cytometer verified that ceils apoptosis after 72 h exposion to costimulate activation of PBMC. Conclusion CD80 can elevate the killing effect of costimulated activation of PBMC with CD28/ CpGODN and on gastric cancerous cell line in vitro. The costimulate activation of PBMC perhaps killed the target cells in two ways：necrosls and apoptosis.

关 键 词：PBMC 单克隆抗体 胃癌 

分 类 号：R567.2[医药卫生—呼吸系统]