CAJ | 학술논문

背景:启动维持并调节活化级联反应,决定了T细胞是活化增殖或转变为无反应状态甚至凋亡.B7分子与CD28的结合能有效协同T细胞受体途径活化T细胞,增强T细胞的增殖活性. 目的:初步分析CD80和CD28/含CpG的寡脱氧核苷酸(CpG containing oligodeoxynucleotides,CpG ODN)共刺激活化的人外周血单个核细胞对胃癌细胞株MKN45的杀伤作用.方法:Ficoll密度梯度离心法分离人外周血单个核细胞,加入白细胞介素2及CD28,CpG ODN共刺激活化培养1～5 d.取MKN45细胞,设立4组:CD28/CpG ODN共刺激组、CD80联合CD28/CpG ODN共刺激组、单纯CD80组、空白对照组.以MTT法检测细胞杀伤率,电镜观察凋亡的MKN45细胞超微结构,流式细胞仪检测MKN45细胞凋亡率.结果与结论:单纯CD80组对MKN45细胞无明显杀伤作用;与CD28/CpG ODN共刺激组比较,CD80联合CD28/CpG ODN共刺激组对MKN45细胞的杀伤作用显著增强(P < 0.05),效应细胞与靶细胞之比达15︰1时即可达到半数杀伤率.电镜下效应细胞作用24 h,MKN45细胞就部分发生坏死,部分细胞可见凋亡.与空白对照组比较,单纯CD80组MKN45细胞凋亡率显著升高(P < 0.01).提示CD80与共刺激活化的外周血单个核细胞合用,可提高活化的外周血单个核细胞对MKN45细胞的靶向性杀伤作用.
배경:계동유지병조절활화급련반응,결정료T세포시활화증식혹전변위무반응상태심지조망.B7분자여CD28적결합능유효협동T세포수체도경활화T세포,증강T세포적증식활성. 목적:초보분석CD80화CD28/함CpG적과탈양핵감산(CpG containing oligodeoxynucleotides,CpG ODN)공자격활화적인외주혈단개핵세포대위암세포주MKN45적살상작용.방법:Ficoll밀도제도리심법분리인외주혈단개핵세포,가입백세포개소2급CD28,CpG ODN공자격활화배양1～5 d.취MKN45세포,설립4조:CD28/CpG ODN공자격조、CD80연합CD28/CpG ODN공자격조、단순CD80조、공백대조조.이MTT법검측세포살상솔,전경관찰조망적MKN45세포초미결구,류식세포의검측MKN45세포조망솔.결과여결론:단순CD80조대MKN45세포무명현살상작용;여CD28/CpG ODN공자격조비교,CD80연합CD28/CpG ODN공자격조대MKN45세포적살상작용현저증강(P < 0.05),효응세포여파세포지비체15︰1시즉가체도반수살상솔.전경하효응세포작용24 h,MKN45세포취부분발생배사,부분세포가견조망.여공백대조조비교,단순CD80조MKN45세포조망솔현저승고(P < 0.01).제시CD80여공자격활화적외주혈단개핵세포합용,가제고활화적외주혈단개핵세포대MKN45세포적파향성살상작용.
BACKGROUND: Maintenance and activation of cascade reaction influence T cell proliferation or transformation into nonreactive state even apoptosis. B7 binding to CD28 effectively activates T cells in combination with T cell receptor pathway, and enhances T cell proliferative activity. OBJECTIVE: To investigate the costimulated activation of peripheral blood mononuclear cells (PBMC) with CD28 and CpG containing oligodeoxynucleotides (CpGODN) MoAb combined with CD80, and its killing effect on human gastric cancerous cell line MKN45 in vitro.METHODS: PBMCs were isolated by Ficoll density gradient centrifugation method, and cocultured with interleukin-2, CD28 and CpGODN MoAb for 1-5 days. MKN45 cells were divided into 4 culture conditions: CD28/CpGODN, CD80 plus CD28/CpG ODN, CD80 alone, and blank control.The killing efficiency was measured by MTT method.The ultramicrostructure of cells was observed by electron microscope. Apoptosis was verified by a flow cytometery. RESULTS AND CONCLUSION: CD80 alone did not display killing effect on MKN45 cells. By MTT method, combination of costimulated activation of PBMC with CD28/CpGODN and CD80 showed enhanced killing effect compared with single therapy (P < 0.05), and the ratio of effector cell and target cell at 15: 1 resulted in half killing efficiency. Electron microscope and flow cytometery verified necrotic or apoptotic cells after 24 hours exposure to costimulated activation. Compared with blank control group, CD80 alone elevated the apoptosis rate of MKN45 cells (P < 0.01). Results from the present study show that CD80 can elevate the killing effect of costimulated activation of PBMC with CD28/CpGODN on MKN45 cells in vitro.