转染增殖细胞核抗原肽表位融合白细胞介素-18基因的树突状细胞对T细胞增殖的影响

目的 观察转染增殖细胞核抗原(PCNA)肽表位、白细胞介素(IL)-18融合基因的树突状细胞(DCs)对T淋巴细胞增殖的诱导作用.方法 实验分为DCs组、空载体组、PCNA肽表位基因转染组、PCNA肽表位融合IL-18基因转染组共4组,分别将空质粒(pIPES-EGFP)、PCNA肽表位质粒[pIPES-EGFP-PCNA(6)]和PCNA肽表位融合IL-18基因质粒[pIRES-EGFP-PCNA(6)/IL-18]通过脂质体转染DCs.每组按照DCs∶T淋巴细胞1∶1、1∶10、1∶20、1∶40、1∶80的比例分为4个浓度梯度,将DCs与T淋巴细胞共培养.采用CCK-8法检测T细胞增殖.结果 各组DCs均能刺激T淋巴细胞增殖,DCs∶T为1∶10时,转染pIPES-EGFP-PCNA(6)/IL-18的DCs刺激指数为2.62,优于DCs组(1.70)、空载体组(1.78)和pIPES-EGFP-PCNA(6)组(1.93).结论 转染pIRES-EGFP-PCNA(6)/IL-18的DCs刺激T淋巴细胞增殖的作用最强,DCs:T为1∶10时最佳.

Abstract：

Objective To investigate the proliferation activity of T lymphocytes induced by dendritic cells modified by proliferating cell nuclear antigen (PCNA)-Epitope and interleukin (IL)-18 genes.Methods The experiment was divided into four groups, including DCs group, empty vector group, PCNAEpitope transfection group and PCNA-Epitope plus IL-18 gene group, and every group was assigned to four EGFP), PCNA-Epitope plasmid [pIRES-EGFP-PCNA(6)] and plasmid loaded PCNA-Epitope and IL-18 gene [pIRES-EGFP-PCNA(6)/IL-18] were transfected into peripheral blood-derived dendritic cells, then cocultured with T lymphocytes respectively. T lymphocytes proliferation was determined by CCK-8 assays.Results The index of stimulation of the pIRES-EGFP-PCNA(6)/IL-18 group was 2. 62 when the ratio transfection group (1. 93). Conclusion DCs treated with the PCNA-Epitope and IL-18 gene could stimulate T lymphocytes proliferation more effectively,and the optimal rio of DCs:T was 1:10.

Effect of dendritic cells modified by PCNA-Epitope and interleukin-18 gene on T lymphocytes proliferation

Objective To investigate the proliferation activity of T lymphocytes induced by dendritic cells modified by proliferating cell nuclear antigen (PCNA)-Epitope and interleukin (IL)-18 genes.Methods The experiment was divided into four groups, including DCs group, empty vector group, PCNAEpitope transfection group and PCNA-Epitope plus IL-18 gene group, and every group was assigned to four EGFP), PCNA-Epitope plasmid [pIRES-EGFP-PCNA(6)] and plasmid loaded PCNA-Epitope and IL-18 gene [pIRES-EGFP-PCNA(6)/IL-18] were transfected into peripheral blood-derived dendritic cells, then cocultured with T lymphocytes respectively. T lymphocytes proliferation was determined by CCK-8 assays.Results The index of stimulation of the pIRES-EGFP-PCNA(6)/IL-18 group was 2. 62 when the ratio transfection group (1. 93). Conclusion DCs treated with the PCNA-Epitope and IL-18 gene could stimulate T lymphocytes proliferation more effectively,and the optimal rio of DCs:T was 1:10.