hbv去甘露聚糖修饰逃逸树突状细胞dcsign免疫识别的研究-study on immune recognition of dc sign in escaping dendritic cells modified by hbv mannan.docxVIP

能是通过调控宿主细胞 MAN1A1、MAN1A2 基因和相应蛋白的表达上调介导的【关键词】HBV，DC，DC-SIGN，Golgi MⅠ，去甘露聚糖修饰，免疫识别Demannosylated hepatitis B virus escapes the immune recognition of DC-SIGN on dendritic cellsDivision ofInfectious diseases,Tongji Hospital of Tongji medical college,HuazhongUniversity of Science and Technology Candidate:Zou Xiaojing Supervisor:Pro.TianDeyingAbstractObjective:To investigate whether HBV infection would increase the activity of Golgi MⅠin HepG2 cell line. The effect of inhibiting HBV demannosylation mediated by Golgi M Ⅰon recognition of DC-SIGN and maturation and activation of DC was observed, to investigate the function of DC-SIGN in HBV infection. The mechanism of demannosylated HBV escaping the immune recognition of DC-SIGN on DC to inhibit the activation of DC was approached as well.Methods:To investigate whether HBV infection would increase the activity of Golgi MⅠHepG2 cells and HepG2.2.15 cells were cultured, and Golgi complexes of these cells were purified by sucrose density gradient centrifugation.The activity of Golgi MⅠin Golgi complexes was detected by colorimetric method.The mRNA expression of MAN1A1、MAN1A2 and MAN1C1 was performed by RT-PCR method.The protein expression of MAN1A1、MAN1A2 and MAN1C1 was performed byWestern blot method.To investigate the function of DC-SIGN in recognition of HBV and activation of DCHepG2.2.15 cells were treated by different concentration of Golgi M Ⅰ inhibitorkifunesine, and the Golgi complexes were purified after 24 hours to detect the activity of Golgi MⅠby colorimetric method.Demannosylated HBV was obtained from the culture medium of HepG2.2.15 cells which had been treated by kifunesine(5ug/ml) for 5 days, HBV DNA was detected by fluorescence quantitative PCR.The peripheral blood mononuclear cells of healthy individual were separated and differentiated to DC by the induction of GM-CSF and IL-4. The morphological characteristics were observed by light microscope and transmission electron microscope.Obtained demanno