白细胞介素18基因转染的肺癌细胞对树突状细胞免疫特性的影响

Effect of lung cancer cells transfected with interleukin 18 gene on the immunological activity of dendritic cells

目的:研究IL-18基因转染的肺癌细胞对树突状细胞(DC)表型和免疫活性的影响。方法:构建含IL-12 P40信号肽序列的分泌型人IL-18表达载体并转染NC I-H460肺癌细胞。从人外周血诱导DC,分为未转染组(NT)、空载体组(PV)、基因转染组(GT)和单纯DC组(PD),用流式细胞仪分别测定未经转染肺癌细胞、空载体转染细胞及基因转染细胞刺激的DC和单纯DC表面CD54、CD80、CD83及CD86的表达。用MTT法测定上述4组DC刺激T细胞增殖的作用。用ELISA法测定上述4组DC培养上清中IL-12的表达量。结果:分泌型IL-18表达载体酶切及测序结果与预期结果一致。IL-18转染细胞表达IL-18融合基因及18 kD蛋白。GT组DC表面分子的表达、刺激T细胞增殖的作用及IL-12分泌量均高于其余3组。结论:IL-18基因转染的肺癌细胞可促进DC表面分子表达,增强DC免疫刺激活性及其IL-12的分泌。

AIM： To study the effect of interleukin 18 gene transfected lung cancer cells on the phenotype and immunological activity of dendritic cells （DC）. METHODS： A secretive IL - 18 expression vector containing IL - 12 P40 signal sequence was constructed and transfected into NCI - H460 lung cancer cells. DC induced from human peripheral blood were divided into 4 groups （ NT, PV, GT and PD）. DC were stimulated by non - transfected NCI - H460 cells, pure vector transfected NCI - H460 cells and IL - 18 transfected NCI - 11460 cells respectively for group NT, PV, GT, and non - stimulated DC for group PD. CD54, CD80, CD83 and CD86 on DC in the 4 groups were detected with flow cytometry. T cell proliferation stimulated by DC in the 4 groups was assayed with MTT method. IL - 12 release in cultured DC supernatant was measured by ELISA. RESULTS： Sequencing result of the secretive IL - 18 was correct. The transfected cells expressed IL - 18 fusion gene and 18 kD IL - 18 protein. DC in GT group expressed more surface molecules than those in other 3 groups. T cell proliferation and IL - 12 secretion in GT group were higher than those in other 3 groups. CONCLUSION： IL- 18 gene transfected NCI -H460 cell increases surface molecule expressions on DC. It also enhances immunological activity and IL - 12 secretion in DC.