6Ckine修饰的树突状细胞促进T淋巴细胞增殖和分化的作用  

6Ckine-Modified Dendritic Cells Promotes Proliferation and Differentiation of T Lymphocytes

作　　者：薛刚[1] 程莹[2] 王培红[1] 曹永宽[1] 田伏洲[1] 黄文林[3] 

机构地区：[1]成都军区总医院全军普通外科中心,成都610083 [2]成都军区总医院内分泌科,成都610083 [3]中山大学肿瘤防治中心肿瘤学国家重点实验室,广州510060

出　　处：《中国生物制品学杂志》2009年第8期743-747,共5页Chinese Journal of Biologicals

基　　金：国家基础研究计划(973计划;2004CB518801);广东省科委重大攻关项目(2003A10902)

摘　　要：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 investigate the effect of dendritic cells （DCs） modified by 6Ckine, a secondary lymphoid organ chemokine, on the proliferation and differentiation of T lymphocytes. Methods Human PBMC-derived DCs were infected with replication-defective recombinant adenovirus （Ad-6Ckine） carrying human 6Ckine gene. The effect on expression of 6Ckine and secretion of cytokines, phagocytic ability, phenotype as well as chemotactic activity to T lymphocytes of Ad-6Ckine-DCs were investigated. Ad-6Ckine-DCs were sensitized with colon cancer LoVo ceils and co-cultured with their autologous T lymphocytes, and the effects on T lymphocyte proliferation and differentiation were studied by ^3H incorporation, RT-PCR and ELISA. Results Little effect on the phagocytic ability of DCs was observed within 24 h after transfection with Ad-6Ckine. 6Ckine was expressed in DCs, which promoted the expressions of CD83 and CCR7 and up-regulated the expression of RANTES. Ad-6Ckine-DCs showed significantly chemotactic activity to T lymphoeytes. The antigen-sensitized Ad-6Ckine-DCs promoted the proliferation of T lymphocyte as well as the expressions of T-bet and IL-2. Conclusion The modification with 6Ckine gene promoted the maturation of DCs at a certain degree and enhanced its capacity in recruiting T lymphoeytes and antigen presentation. Ad-6Ckine-DC promoted T lymphocyte proliferation and induced Thl polarization, which might be a candidate of tumor vaccine.

关 键 词：次级淋巴组织趋化因子 树突状细胞 T淋巴细胞 增殖 分化 

分 类 号：R373.1[医药卫生—病原生物学]