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山东大学学报(医学版)

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pcDNA3-HBV瞬时转染对树突状细胞的影响

高立芬1,孙汶生1,马春红1,张利宁1,梁晓红1,王晓燕1,郭春1,陈有海2   

  1. 1. 山东大学医学院免疫学研究所, 山东 济南 250012;2. Department of Pathology and Laboratory Medicine,University of Pennsylvania, PA19104, USA
  • 收稿日期:2006-05-06 修回日期:1900-01-01 出版日期:2007-04-24 发布日期:2007-04-24
  • 通讯作者: 孙汶生

Effects of transient transfection of pcDNA3-HBV on dendritic cells

GAO Li-fen1,SUN Wen-sheng1,MA Chun-hong1,ZHANG Li-ning1,LIANG Xiao-hong1,WANG Xiao-yan1,GUO Chun1,CHEN You-hai2   

  1. 1.Institute of Immunology, School of Medicine, Shandong University, Jinan 250012, Shandong, China;2. Department of Pathology and Laboratory Medicine,University of Pennsylvania, PA19104, USA
  • Received:2006-05-06 Revised:1900-01-01 Online:2007-04-24 Published:2007-04-24
  • Contact: SUN Wen-sheng

摘要: 目的:研究HBV感染对树突状细胞功能的影响,探讨慢性乙型肝炎患者免疫耐受发生的机制。方法:利用脂质体转染的方法,以携载HBV全基因的真核表达载体pcDNA3-HBV及对照空载体pcDNA3质粒DNA分别转染小鼠骨髓细胞,以细胞因子GM-CSF、IL-4及TNF-α诱导培养树突状细胞,RT-PCR检测转染后HBV PreS1 mRNA的表达。流式细胞术检测转染前后树突状细胞表面分子CD80的表达,3HTdR掺入法观察其刺激同种异体淋巴细胞增殖的能力,流式细胞术进一步检测NF-κB RelA蛋白的表达。结果:pcDNA3-HBV瞬时转染后树突状细胞CD80表达降低,其刺激同种异体淋巴细胞增殖的能力下降,NF-κB RelA蛋白表达显著减少。结论: HBV可抑制树突状细胞的分化成熟,降低其刺激T淋巴细胞增殖的功能,这可能是慢性乙型肝炎患者免疫耐受的机制之一。

Abstract: Objective: To study the role of HBV infection on the function of dendritic cells (DCs) and explore the mechanism of immune tolerance in chronic B hepatitis. Methods: The bone marrow cells were isolated, cultured and transfected transiently with pcDNA3-HBV or pcDNA3 plasmid DNA, and then they were cultured under the stimulation of GM-CSF, IL-4, TNF-α to be induced to become mature dendritic cells. RT-PCR was performed to detect the expression of PreS1 mRNA. FCM was used to detect the expression of CD80, NF-κB. 3H-TdR adulteration assay was used to detect the potential for stimulating the proliferation of allogenic lymphocytes. Results: After being transfected with pcDNA3-HBV, mature DCs had the expression of PreS1 mRNA, but had lower level of superficial marker CD80 and transcriptional factor NF-κB, compared with pcDNA3-transfected control cells; the potential for stimulating the proliferation of allogenic lymphocytes was also decreased. Conclusion: Transient transfection of pcDNA3-HBV could decrease the function of DCs directly by inhibiting the maturation of DCs, which may be one of the mechanisms of immune tolerance in chronic B hepatitis.

Key words: Hepatitis B virus, Transfection, Dendritic cells

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  • R392.12
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