HCV-NS3重组腺病毒转染树突状细胞体内诱导抗原特异性Th1应答的研究  

Th1 cellular immune response elicited by recombinant adenovirus transduced dendritic cells for HCV-NS3 in vivo

作　　者：向明[1] Jens Encke 

机构地区：[1]华中科技大学同济药学院药理室,武汉430030 [2]德国海德堡大学临床医学研究所

出　　处：《中国免疫学杂志》2006年第10期921-924,共4页Chinese Journal of Immunology

摘　　要：目的:探讨表达丙型肝炎病毒(HCV)非结构蛋白(NS3)基因的重组腺病毒转染树突状细胞体内诱导特异性Th1细胞免疫应答。方法:分离培养小鼠骨髓树突状细胞,制备表达HCV-NS3蛋白的重组腺病毒转染树突状细胞(DC-AdNS3)疫苗,采用流式细胞术和免疫印迹法分析鉴定细胞及抗原蛋白表达。采用腹腔注射途径免疫接种BALB/c小鼠两次,每次间隔10天,3×105细胞/次。末次接种10天后,采用ELISPOT法和ELISA法分别测定脾NS3特异性分泌IFN-γ的T细胞反应以及细胞因子水平。结果:重组腺病毒转染DC可刺激DC成熟,同时可在DC内成功表达NS3蛋白。小鼠两次接种DC-AdNS3产生明显升高的分泌IFN-γ的T细胞反应(P<0·01),脾T细胞悬液内可测得高水平的IL-2和IFN-γ(P<0·01)以及显著降低的IL-10(P<0·05)。结论:DC-AdNS3疫苗可在BALB/c小鼠体内激发产生抗原特异性的Th1细胞免疫应答,为抗HCV感染的疫苗研究提供参考依据。Objective： The aim of this study was to explore the vaccine potential of adenovirus transduced DC encoding for HCV nonstructural protein NS3 in a murine model. Methods： Dendritic cells were isolated from bone marrow of BALB/c mice and cultivated with cytokine cocktail for 7 days. Recombinant adenovirus was added to DC cultures at day 7 ,at a titration of 2 × 10^9 pfu/10^6 DC/well. FACS and Western blot techniques were used to identify the DC mature state and the protein expression, respectively. BALB/c mice were primed/boosted immunized with DC based vaccines at 3 × 10^5 cells for one mouse at a 10 day interval. 10 days after the second immunization, cytokines secretion and IFN-γ generating T cells response were evaluated by ELISA and ELL/SPOT analysis. Results： Our current data indicate that AdNS3 can stimulate the maturation of DC and efficiently express authentic HCV nonstructural proteins in infected cells. BALB/c mice inoculated with AdNS3 transduced DC twice generated strong IFN-γ secreting T cells response（ P 〈 0. 01 ） and high secretion of IL-2 and IFN-γ（ P 〈 0.01 ） and decreased level of IL-10 （ P 〈 0. 05 ）. Conclusion ： Antigen specific immune response can he induced in BALB/c mice immunized with AdNS3 transduced DC, which provide the facts for application in HCV prevention and therapeutic settings.

关 键 词：丙肝非结构蛋白 腺病毒 树突状细胞 TH1免疫应答 

分 类 号：R392[医药卫生—免疫学]