HBV对健康产妇脐带血浆细胞样树突状细胞Toll样受体9、干扰素调节因子7、干扰素α表达的影响

彭建平; 孙克伟; 银思涵; 伍玉南

doi:10.3969/j.issn.1001-5256.2016.05.016

HBV对健康产妇脐带血浆细胞样树突状细胞Toll样受体9、干扰素调节因子7、干扰素α表达的影响

DOI: 10.3969/j.issn.1001-5256.2016.05.016

湖南中医药大学第一附属医院传染科

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- 出版日期: 2016-05-20

Effects of hepatitis B virus on the expression of Toll-like receptor 9,interferon regulatory factor-7,and interferon-α in plasmacytoid dendritic cells in the umbilical cord blood of healthy parturients

Department of Infectious Diseases, The First Affiliated Hospital of Hunan University of Chinese Medicine

摘要

摘要:
目的观察HBV对体外培养的健康产妇脐带血浆细胞样树突状细胞（pDC）Toll样受体（TLR）9、干扰素调节因子（IRF）7、干扰素（IFN）α表达的影响。方法体外培养Hep G 2.2.15细胞,收集各孔Hep G 2.2.15培养上清,超离心HBV病毒颗粒,PCR荧光探针法检测HBV DNA浓度。将培养第7天的pDC与不同载量的HBV（1×105拷贝/ml、1×106拷贝/ml、1×107拷贝/ml和空白对照）共培养24 h。随后每组均加入终浓度为2μg/ml的Cp G ODN 2216继续培养24 h。Real Time-PCR检测pDC中TLR9、IRF7mRNA的表达,Western Blot检测TLR9、IRF7蛋白表达,ELISA法测IFNα水平。计量资料多组间比较采用方差分析,进一步两两比较采用SNK-q检验;对HBV DNA拷贝数取常用对数,相关指标与HBV DNA的常用对数值用Pearson直线相关分析法。结果HBV 1×106拷贝/ml组和1×107拷贝/ml组TLR9 mRNA、IRF7 mRNA、pDC TLR9和IRF7蛋白、IFNα的表达水平明显低于空白对照...
- 肝炎病毒,乙型 /
- 树突细胞 /
- Toll样受体9 /
- 干扰素调节因子7 /
- 干扰素α
Abstract:
Objective To investigate the effects of hepatitis B virus( HBV) on the expression of Toll- like receptor 9( TLR9),interferon regulatory factor- 7( IRF- 7),and interferonα( IFNα) in plasmacytoid dendritic cells( pDCs) cultured in vitro in the umbilical cord blood of healthy parturients. Methods Hep G 2. 2. 15 cells were cultured in vitro,and the culture supernatants of Hep G 2. 2. 15 in each well were collected. HBV particles were obtained by ultracentrifugation,and HBV DNA concentration was measured by fluorescent- probe PCR. pDCs( on day 7 of culture) and HBV( 1 × 105 copies / ml,1 × 106 copies / ml,1 × 107 copies / ml,and blank control) were co- cultured for 24 hours,and then cultured continuously for another 24 hours with Cp G ODN 2216 at a final concentration of 2 μg / ml. The mRNA expression of TLR9 and IRF- 7 in pDCs was measured by real- time PCR,the protein expression of TLR9 and IRF- 7 in pDCs was measured by Western blot,and the level of IFNα was measured by ELISA. The analysis of variance was used for comparison of continuous data between multiple groups,and the SNK- q test was used for comparison between any two groups; the common logarithms of HBV DNA copies were obtained,and the linear regression analysis was performed to investigate the correlation between related indicators and the common logarithms of HBV DNA copies. Results Compared with the blank control group and the HBV 1 × 105 copies / ml group,the HBV 1 × 106 copies / ml and HBV 1 × 107 copies / ml groups had significantly lower mRNA expression of TLR9 and IRF- 7,significantly lower protein expression of TLR9 and IRF- 7,and significantly lower expression of IFNα( all P < 0. 05). The inhibitory effects of HBV on the mRNA expression of TLR9 and IRF- 7,the protein expression of TLR9 and IRF- 7,and the expression of IFNα in the culture supernatants of pDCs rose with the increasing HBV DNA load( r =- 0. 729,- 0. 761,- 0. 657,- 0. 703,and- 0. 803,all P < 0. 05). Conclusion After in vitro treatment with HBV,the mRNA and protein expression of TLR9 and IRF- 7 in normal pDCs is reduced,leading to reduced secretion of downstream IFNα. HBV can inhibit the function of normal pDCs and cause persistent HBV infection.
- hepatitis B virus /
- dendritic cells /
- Toll-like receptor 9 /
- interferon regulatory factor-7 /
- interferon-alpha

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参考文献(14)

[1]ZHANG X,LEPELLEY A,AZRIA E,et al.Neonatal plasmacytoid dendritic cells(p DCs)display subset variation but can elicit potent anti-viral innate responses[J].PLoS One,2013,8(1):e52003.

[2]LOMBARDI VC,KHAIBOULLINA SF.Plasmacytoid dendritic cells of the gut:relevance to immunity and pathology[J].Clin Immunol,2014,153(1):165-177.

[3]O'BRIEN M,MANCHES O,BHARDWAJ N.Plasmacytoid dendritic cells in HIV infection[J].Adv Exp Med Biol,2013,762:71-107.

[4]DENTAL C,FLORENTIN J,AOUAR B,et al.Hepatitis C virus fails to activate NF-κB signaling in plasmacytoid dendritic cells[J].J Virol,2012,86(2):1090-1096.

[5]PENG JP,SUN KW,YIN SS,et al.In vitro culture and characterization of human umbilical cord blood-derived plasmacytoid dendritic cell subsets[J].J Clin Hepatol,2015,31(11):1862-1866.(in Chinese)彭建平,孙克伟,银丝丝,等.人脐带血浆细胞样树突状细胞亚群的体外诱导及鉴定[J].临床肝胆病杂志,2015,31(11):1862-1866.

[6]LIU MC,YU M,ZHANG NL,et al.Dynamic analysis of hepatitis B virus DNA and its antigens in 2.2.15 cells[J].J Viral Hepat,2004,11(2):124-129.

[7]DREUX M,GARAIGORTA U,BOYD B,et al.Short-range exosomal transfer of viral RNA from infected cells to plasmacytoid dendritic cells triggers innate immunity[J].Cell Host Microbe,2012,12(4):558-570.

[8]SAJADI SM,MIRZAEI V,HASSANSHAHI G,et al.Decreased expressions of Toll-like receptor 9 and its signaling molecules in chronic hepatitis B virus-infected patients[J].Arch Pathol Lab Med,2013,137(11):1674-1679.

[9]VINCENT IE,ZANNETTI C,LUCIFORA J,et al.Hepatitis B virus impairs TLR9 expression and function in plasmacytoid dendritic cells[J].PLoS One,2011,6(10):e26315.

[10]XIE Q,SHEN HC,JIA NN,et al.Patients with chronic hepatitis B infection display deficiency of plasmacytoid dendritic cells with reduced expression of TLR9[J].Microbes Infect,2009,11(4):515-523.

[11]WOLTMAN AM,OP den BROUW ML,BIESTA PJ,et al.Hepatitis B virus lacks immune activating capacity,but actively inhibits plasmacytoid dendritic cell function[J].PLoS One,2011,6(1):e15324.

[12]MA J,HHANG C,YAO X,et al.Inhibition of hepatitis B virus and induction of hepatoma cell apoptosis by ASGPR-directed delivery of shRNAs[J].PLoS One,2012,7(10):e46096.

[13]SHAHRAKYVAHED A,SANCHOOLI J,SANADGOL N,et al.TLR9:an important molecule in the fight against hepatitis B virus[J].Postgrad Med J,2014,90(1065):396-401.

[14]FATHALLAH I,PARROCHE P,GRUFFAT H,et al.EBV latent membrane protein 1 is a negative regulator of TLR9[J].J Immunol,2010,185(11):6439-6447.

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