论文部分内容阅读
目的:探讨CD40L~+肺癌细胞抗原负载由人外周血单个核细胞(peripheral blood mononuclear cell,PBMC)诱导的DC对同源肺癌的免疫增强作用及其机制。方法:常规方法从正常成人外周血中分离PBMC、诱导分化成DC,以重组载体p DNA3.1-CD40L~+转染A549肺癌细胞,制备已转染肺癌细胞抗原对DC进行负载刺激,通过流式细胞术检测DC的CD83、CD86及HLA-DR分子表达,ELISA法检测DC的IL-12分泌水平,再以MTT法检测DC对同源T淋巴细胞增殖状态以及A549肺癌细胞增殖影响,并与空载转染的A549细胞抗原诱导DC的相同功能进行对比。结果:将PBMC成功诱导分化出成熟的DC。重组CD40L诱导组DC的CD83、CD86及HLA-DR分子表达水平明显高于空载组[(4.78±1.5)%vs(3.38±1.5)%、(9.79±5.27)%vs(5.53±2.17)%和(11.84±8.31)%vs(5.37±5.48)%,均P<0.05];IL-12分泌水平也高于空载组和未转染组(P<0.05)。CD40L~+DC可促进T淋巴细胞的增殖(P<0.05),并导致同源T细胞对肺癌A549细胞增殖的抑制作用强于对照组、未转染组和空载组(P<0.05)。结论:CD40L转染肺癌细胞抗原诱导成熟的DC可以增强同源肺癌细胞的特异免疫能力。
Objective: To investigate the immune-enhancing effect of CD40L ~ + lung cancer cell antigen on lung cancer induced by peripheral blood mononuclear cell (PBMC) and its mechanism. Methods: PBMCs were isolated from normal adult peripheral blood by routine methods and induced to differentiate into DC. A549 lung cancer cells were transfected with recombined vector p DNA3.1-CD40L ~ +. The transfected lung cancer cell antigens were stimulated by DCs. The expression of CD83, CD86 and HLA-DR in DCs were detected by cytometry. The levels of IL-12 in DCs were detected by ELISA. The proliferation of DCs and the proliferation of A549 lung cancer cells were detected by MTT assay. The same function induced by DCs transfected with A549 cells was compared. Results: PBMCs were successfully induced to differentiate into mature DCs. The expression levels of CD83, CD86 and HLA-DR in DCs induced by recombinant CD40L were significantly higher than those in control group [(4.78 ± 1.5)% vs (3.38 ± 1.5%, (9.79 ± 5.27)% vs (5.53 ± 2.17% And (11.84 ± 8.31)% vs (5.37 ± 5.48)%, all P <0.05]. The level of IL-12 secretion was also higher than that of the control group (P <0.05). CD40L ~ + DC promoted the proliferation of T lymphocytes (P <0.05), and caused the inhibitory effect of homologous T cells on the proliferation of lung cancer A549 cells more than that of control group, untransfected group and no-load group (P <0.05). Conclusion: CD40L transfection of lung cancer cell antigen induced mature DC can enhance the specific immunity of lung cancer cells.