CD40L^+肺癌细胞抗原负载DC诱导同源肺癌免疫

Dendritic cells,pulsed with CD40L^+ lung cancer antigens,induce homologous lung cancer immunity

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 explore the immunological enhancement effect of dendritic cells （DCs） derived from peripheral blood mononuclear cells （PBMCs） pulsed CD40L＋ lung cancer cell antigen , on homologous lung cancer. Methods： PBMCs were isolated from peripheral blood of adults by conventional methods, then induced differentiation into DCs; recombinant CD40L＋A549 cell antigen was used for DC maturation stimulation. The expressions of CD83,CD86 and HLA-DR in DCs were detected by Flow Cytometry, IL-12 secretion was detected by ELISA; the effects of CD40L＋ lung cancer cell antigen pulsed DCs on proliferation of homologous lymphocyte as well as A549 cells were examined by MTT, in addition, this effect was compared to the effect of DCs that pulsed by empty vector plasmid transfected A549 cell antigen. Results： Mature DCs were successfully induced from PBMCs. The expressions of CD83, CD86 and HLA-DR in CD40L＋ A549-pulsed DC group were significantly higher than those in empty 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]; and the secretion of IL-12 in CD40L＋ A549-pulsed DC group was also higher than those in empty group and untransfected group （P〈0.05）. In the mean while, CD40L-pulsed DCs significantly promoted the proliferation of lymphocytes （P〈0.05）, and its activated homologous T cells had a significantly higher inhibitory effect on A549 cell proliferation （P〈0.05）. Conclusion： DC pulsed by the tumor antigens from the reconstitution CD40L transfected A549 cells could enhance its specific immunity capacity against homologous lung cancer cells.