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目的研究次级淋巴组织趋化因子(6Ckine)修饰的树突状细胞(DC)对T淋巴细胞增殖和分化的影响。方法用携带人6Ckine基因的重组复制缺陷型腺病毒(Ad-6Ckine)感染人外周血单个核细胞来源的DC,检测Ad-6Ckine-DC对6Ckine的表达及细胞因子分泌的影响,并观察其吞噬功能和表型的变化及对自身T淋巴细胞的趋化作用。用结肠癌LoVo细胞抗原致敏Ad-6Ckine-DC,将该DC与自身T淋巴细胞共同培养,分别用3H掺入法、RT-PCR和ELISA检测Ad-6Ckine-DC对T淋巴细胞增殖和分化的影响。结果在Ad-6Ckine转染后24h内,DC的吞噬功能几乎不受影响。转染的6Ckine基因能在DC中表达,表达的6Ckine能促进其表达CD83和CCR7,上调RANTES的表达。Ad-6Ckine-DC对自身T淋巴细胞有明显的趋化作用,抗原致敏的Ad-6Ckine-DC能显著促进T淋巴细胞的增殖,并增强其表达T-bet和IL-2的能力。结论6Ckine基因的修饰能在一定程度上促进DC的成熟,并募集T淋巴细胞于DC周围,有利于DC向T淋巴细胞传递抗原和第二信息,增强DC促进T淋巴细胞增殖的作用并使其向Th1分化,诱导细胞免疫,将成为制备肿瘤疫苗的一种良好选择。
Objective To study the effect of dendritic cells (DCs) modified by secondary chemokines (6Ckine) on the proliferation and differentiation of T lymphocytes. METHODS: Human peripheral blood mononuclear cell-derived DCs were infected with recombinant replication-defective adenovirus carrying human 6Ckine gene (Ad-6Ckine) to detect the effect of Ad-6Ckine-DC on 6Ckine expression and cytokine secretion. Function and phenotype changes and chemotaxis to their own T lymphocytes. Ad-6Ckine-DC was sensitized with colon cancer LoVo cell antigen and co-cultured with its own T lymphocytes. 3H-incorporation, RT-PCR and ELISA were used to detect the proliferation and differentiation of T lymphocytes Impact. Results Within 24 hours after Ad-6Ckine transfection, phagocytosis of DC was almost unaffected. Transfected 6Ckine gene can be expressed in DC, the expression of 6Ckine can promote its expression of CD83 and CCR7, up-regulate the expression of RANTES. Ad-6Ckine-DC has obvious chemotactic effect on T lymphocytes. Antigen-sensitized Ad-6Ckine-DC can significantly promote the proliferation of T lymphocytes and enhance the expression of T-bet and IL-2. Conclusion The modification of 6Ckine gene can promote the maturation of DC to some extent and recruiting T lymphocytes around DC, which is helpful for DC to transmit antigens and second information to T lymphocytes and enhance the effect of DC on T lymphocyte proliferation To Th1 differentiation, induction of cellular immunity, will become a good choice for the preparation of tumor vaccines.