HBcAg对健康人及乙肝病毒携带者外周血单个核细胞刺激产生IFN-α影响的研究
摘要
目的:构建表达乙肝病毒核心抗原(HBcAg)的复制缺陷型重组腺病毒载体,将其分别转染健康人及乙肝病毒携带者外周血单个核细胞,观察其刺激产生α干扰素水平并与TLR9的天然配体CpG ODN 2006刺激效果进行比较,以初步探讨乙型肝炎病毒感染免疫识别状态及乙肝病毒感染慢性化机制。
方法:扩增乙肝病毒(HBV)C基因片段,亚克隆到穿梭质粒pAdTrack-CMV上,与5型腺病毒骨架质粒pAdeasy-1共同电转化大肠杆菌BJ5183内进行同源重组,经卡那霉素抗性筛选和酶切鉴定筛选出携带(HBV)C区基因的重组腺病毒载体,用脂质体包裹PacⅠ酶切线性化的重组质粒,转染到293T细胞内进行重组腺病毒的包装,获得具有感染能力的重组腺病毒颗Ad-HBc。应用密度梯度离心法从健康人(HBsAg阴性)及乙肝病毒携带者外周静脉血中分离出单个核细胞,分为四组加入干预因素:Ad-HBc组、CpG ODN 2006组、Ad-HBc+CpG ODN 2006组和空白对照组,孵育48小时后采用ELISA检测各组细胞培养上清中IFN-α水平。
结果:在健康人中,与空白对照组比较,Ad-HBc+CpG ODN 2006组、Ad-HBc组和CpG ODN 2006组中IFN-α均有明显升高(P<0.05),Ad-HBc+CpG ODN 2006组升高最明显,Ad-HBc组次之,CpG ODN 2006组最低。在携带者中,与空白对照组比较,Ad-HBc+CpG ODN 2006组升高较明显,CpG ODN 2006组次之,Ad-HBc组最低(P<0.05),且Ad-HBc组水平明显低于健康人同组水平(P<0.05)。
结论:携带乙肝核心抗原基因复制缺陷型重组腺病毒感染健康人外周血单个核细胞能诱导增加健康人PBMC产生IFN-α的能力,且刺激效果优于TLR9的配体CpG ODN 2006。而在乙肝病毒携带者中,重组腺病毒感染外周血单个核细胞产生IFN-α水平低于健康人,提示与HBV持续感染造成外周血pDC功能下降有关,为进一步研究HBV持续感染的免疫机制提供了思路。
Objective:To construct recombinant adenoviral vector carrying HBcAg gene by homologous recombination in bacteria and to transfect human peripheral blood mononuclear cells from healthy subjects and HBV carriers, analyze the value of IFN-αin culture supernatants by ELISA and compare to the results of CpG ODN 2006 (ligand of TLR9) in purpose of investigating the mechanism of hepatitis B virus infection recognition in immune system.
Methods:HBV C genes were amplified by PCR and were cloned to adenoviral shuttle plasmid pAdTrack-CMV. Then the resultant pAdTrack-CMV-HBc was cotransfected into BJ5183 bacteria with the plasmid pAdeasy-1. The adenoviral plasmid carrying HBV C gene (pAd-HBc) was generated with homologous recombination in bacteria and the adenoviruses were produced in 293 T cells. The recombinant adenoviruses Ad- HBc were obtained. With the method of density gradient centrifugation,isolating peripheral blood mononuclear cells from healthy subjects(HBsAg negative) and HBV carriers were divided into four groups. Three groups was stimulated respectively by Ad-HBc+CpG ODN 2006、Ad-HBc、CpG ODN 2006 and the fourth was control group. After 48 hours, the value of IFN-αin culture supernatants was detected by ELISA.
Results:In healthy subjects, compared with control, the concentration of IFN-αin Ad-HBc+CpG ODN 2006 group、Ad-HBc group and CpG ODN 2006 group were all elevated(P<0.05),Ad-HBc+CpG ODN 2006 group increased significantly,Ad-HBc group followed,CpG ODN 2006 group the lowest. In HBV carriers, compared with control, Ad-HBc + CpG ODN 2006 group increased slightly ,CpG ODN 2006 group followed, Ad-HBc group the lowest (P<0.05).The IFN-αconcentration of Ad-HBc in HBV carriers is significantly lower than healthy subjects(P<0.05).
Conclusions:Replication- deficient recombinant adenoviral vector carrying HBcAg gene infected human peripheral blood mononuclear cells from healthy subjects, increased ability of pDC can be induced to produce IFN-α.The stimulated result was more effective than the ligand CpG ODN 2006 of TLR9. In HBV carriers, the adenovirus infection of peripheral blood mononuclear cells were disable to produce IFN-αmore than CpG ODN 2006,the result may involve in dysfunction of pDCs caused by persistent infection of HBV.
方法:扩增乙肝病毒(HBV)C基因片段,亚克隆到穿梭质粒pAdTrack-CMV上,与5型腺病毒骨架质粒pAdeasy-1共同电转化大肠杆菌BJ5183内进行同源重组,经卡那霉素抗性筛选和酶切鉴定筛选出携带(HBV)C区基因的重组腺病毒载体,用脂质体包裹PacⅠ酶切线性化的重组质粒,转染到293T细胞内进行重组腺病毒的包装,获得具有感染能力的重组腺病毒颗Ad-HBc。应用密度梯度离心法从健康人(HBsAg阴性)及乙肝病毒携带者外周静脉血中分离出单个核细胞,分为四组加入干预因素:Ad-HBc组、CpG ODN 2006组、Ad-HBc+CpG ODN 2006组和空白对照组,孵育48小时后采用ELISA检测各组细胞培养上清中IFN-α水平。
结果:在健康人中,与空白对照组比较,Ad-HBc+CpG ODN 2006组、Ad-HBc组和CpG ODN 2006组中IFN-α均有明显升高(P<0.05),Ad-HBc+CpG ODN 2006组升高最明显,Ad-HBc组次之,CpG ODN 2006组最低。在携带者中,与空白对照组比较,Ad-HBc+CpG ODN 2006组升高较明显,CpG ODN 2006组次之,Ad-HBc组最低(P<0.05),且Ad-HBc组水平明显低于健康人同组水平(P<0.05)。
结论:携带乙肝核心抗原基因复制缺陷型重组腺病毒感染健康人外周血单个核细胞能诱导增加健康人PBMC产生IFN-α的能力,且刺激效果优于TLR9的配体CpG ODN 2006。而在乙肝病毒携带者中,重组腺病毒感染外周血单个核细胞产生IFN-α水平低于健康人,提示与HBV持续感染造成外周血pDC功能下降有关,为进一步研究HBV持续感染的免疫机制提供了思路。
Objective:To construct recombinant adenoviral vector carrying HBcAg gene by homologous recombination in bacteria and to transfect human peripheral blood mononuclear cells from healthy subjects and HBV carriers, analyze the value of IFN-αin culture supernatants by ELISA and compare to the results of CpG ODN 2006 (ligand of TLR9) in purpose of investigating the mechanism of hepatitis B virus infection recognition in immune system.
Methods:HBV C genes were amplified by PCR and were cloned to adenoviral shuttle plasmid pAdTrack-CMV. Then the resultant pAdTrack-CMV-HBc was cotransfected into BJ5183 bacteria with the plasmid pAdeasy-1. The adenoviral plasmid carrying HBV C gene (pAd-HBc) was generated with homologous recombination in bacteria and the adenoviruses were produced in 293 T cells. The recombinant adenoviruses Ad- HBc were obtained. With the method of density gradient centrifugation,isolating peripheral blood mononuclear cells from healthy subjects(HBsAg negative) and HBV carriers were divided into four groups. Three groups was stimulated respectively by Ad-HBc+CpG ODN 2006、Ad-HBc、CpG ODN 2006 and the fourth was control group. After 48 hours, the value of IFN-αin culture supernatants was detected by ELISA.
Results:In healthy subjects, compared with control, the concentration of IFN-αin Ad-HBc+CpG ODN 2006 group、Ad-HBc group and CpG ODN 2006 group were all elevated(P<0.05),Ad-HBc+CpG ODN 2006 group increased significantly,Ad-HBc group followed,CpG ODN 2006 group the lowest. In HBV carriers, compared with control, Ad-HBc + CpG ODN 2006 group increased slightly ,CpG ODN 2006 group followed, Ad-HBc group the lowest (P<0.05).The IFN-αconcentration of Ad-HBc in HBV carriers is significantly lower than healthy subjects(P<0.05).
Conclusions:Replication- deficient recombinant adenoviral vector carrying HBcAg gene infected human peripheral blood mononuclear cells from healthy subjects, increased ability of pDC can be induced to produce IFN-α.The stimulated result was more effective than the ligand CpG ODN 2006 of TLR9. In HBV carriers, the adenovirus infection of peripheral blood mononuclear cells were disable to produce IFN-αmore than CpG ODN 2006,the result may involve in dysfunction of pDCs caused by persistent infection of HBV.
引文
[1] Jung MC, Pape GR. Immunology of hepatitis B infection. Lancet Infect Dis 2002; 2:43-50.
[2] Lok, A.S, Heathcote, E.J, Hoofnagle, J.H. Management of hepatitis B: 2000-Summary of a workshop. Gastroenterology, 2001;120(7):1828-1853.
[3] Lau DT, Everhart J, Kleiner DE, Park Y, Vergalla J, Schmid P, Hoofnagle JH. Long-term follow-up of patients with chronic hepatitis B treated with interferon alfa. Gastroenterology 1997; 113: 1660-1667
[4] Patricia Fitzgerald-Bocarsly. Plasmacytoid dendritic cells and type I IFN:50 years of convergent history. Cytokine & Growth Factor Reviews 2008 19:3-19
[5] Shizuo Akira, Satoshi Uematsu, Pathogen Recognition and Innate Immunity. Cell.2006;124:783-801
[6]中华医学会肝病学分会,中华医学会感染病学分会.慢性乙型肝炎防治指南.2005
[7]黄呈辉,欧阳玲等,携带乙肝核心抗原基因复制缺陷型重组腺病毒的高效制备和表达。中国医学工程2006 ;14:616-621
[8] Stefan Bauer,Carsten J. Kirschning. Human TLR9 confers responsiveness to bacterial DNA via species-specific CpG motif recognition. PNAS .2001; 98(16) :9237–9242
[9] Ganem,D. and A.M. Prince.Hepatitis B virus infection-natural history and clinical consequences.N Engl J Med.2004;350(11):1118-1129
[10] Maria-Christina Jung ,Gerd R Pape. Immunology of hepatitis B infection. Lancet Infectious Diseases .2002 ;2:43-50
[11] Stefan Wieland,Robert Thimme. Genomic analysis of the host response to hepatitis B virus infection. PNAS.2004;101:6669-6674
[12] Luke AJ O’Neill. How Toll-like receptors signal: what we know and what we don’t know. Current Opinion in Immunology. 2006,;18:3–9
[13] Kenya Honda,Hideyuki Yanai.IRF-7 is the master regulator of type-I interferon-dependent immune responses. Nature .2005;434:772-777
[14] Norimitsu Kadowaki , Yong-Jun Liu. Natural Type I Interferon-Producing Cells as a Link Between Innate and Adaptive Immunity. Human Immunology.2002;63:1126–1132
[15] Kelli McKenna,Anne-Sophie Beignon. Plasmacytoid Dendritic Cells:Linking Innate and Adaptive Immunity.Journal of Virology.2005;79(1):17-27
[16] Gregory M. Barton,Jonathan C. Kagan. A cell biological view of Toll-like receptor function: regulation through compartmentalization. Nature Reviews Immunology. 2009;9:535-542
[17] Y Kumagai,O Takeuchi,S Akira. TLR 9 as a key receptor for the recognition of DNA. Advanced Drug Delivery Reviews.2008;60(7):795-804
[18] Jennifer Lund, Ayuko Sato. Toll-like Receptor 9-mediated Recognition of Herpes Simplex Virus-2 by Plasmacytoid Dendritic Cells. J. Exp. Med. 2003 ;198:513-520
[19] Jorg Schlender,Veit Hornung. Inhibition of toll-like receptor 7 and 9 mediated alpha/beta interferon production in human plasmacytoid dendritic cells by respiratory syncytial virus and measles virus. Journal of virology.2005;79(9):5507-5515
[20] Isogawa M. Robek MD.Furuich Y. Toll-like receptor signaling inhibits hepatitis B virus replication in vivo.J Virol.2005;79(11):7269-7272
[21] Fanning SL,George TC, Feng D, et al.Receptor cross-linking on human plasmacytoid dendritic cells leads to the regulation of IFN-alpha production.J Immunol 2006; 177:5829-39
[22] Feldman SB, Milone MC,Kloser P,Fitzgerald-Bocarsly P.Functional deficiencies in two distinct IFN-a producing cell populations in PBMC from human immunodeficiency virus seropositive patients.J Leu Biol.1995;57:214-20
[23] Heather W.Aging impairs IFN regulatory factor 7 up-regulation in plasmacytoid dendritic cells during TLR 9 activation. The Journal of Immunology.2008;181(10):6747-6756
[24] Chaperot L, Bendriss N, Manches O, Gressin R,et al. Identification of a leukemic counterpart of the plasmacytoid dendritic cells. Blood .2001;97:3210-7
[25] Heathcote J, Mchutchison J,Lee S,et al.A pilot study of C-1899 T-cell vaccine in subjects chronically infected with hepatitis B virus. The CY 1899 T-cell vaccine study group.Hepatology.1999;30:531-6
[26] Bertoletti A, Chisari FV, Penna A,et al. Definition of a minimal optimal cytotoxic T-cell epitope within the hepatitis B virus nucleocapsid protein. J Virol 1993;67:2376-80
[27] Ferrari C, Penna A,Bertoletti A,et al.Cellular immune response to hepatitis B virus-encoded antigens in acute and chronic hepatitis B virus infection.J Immunol 1990;145:3442-9
[28] Bertoletti A, Southwood S,Chesnut R,et al.Molecular features of the hepatitis B virus mucleocapsid T-cell epitopes 18-27:interaction with HLA and T-cell receptor. Hepatology.1997;26:1027-34
[29] Milich DR, Schodell F,Hughes JL,et al.The hepatitis B virus core and e antigens elicit different Th cell subsets: antigen structure can affect Th cell phenotype.J Virol 1997;71:2192-201
[30] Milich DR.Immune response to hepatitis B virus:infection,animal model,vaccine.Viral Hepat Rev 1997;3:63-103
[31] Qing Xie,Huai-Cheng Shen,et al. Patients with chronic hepatitis B infection displaydeficiency of plasmacytoid dendritic cells with reduced expression of TLR9. Microbes and Infection. 2009 11:515-523
[32] Jian ZHOU, Yuancheng HUANG. Expression of Toll-like Receptor 9 in Peripheral Blood Mononuclear Cells from Patients with Different Hepatitis B and C Viral Loads. J Huazhong Univ Sci Technol. 2009 29(3):313-317
[33] Wang FS, Xing LH, Liu MX, et al. Dysfunction of peripheral blood dendritic cells from patients with chronic hepatitis B virus infection. World J Gastroenterol 2001;7:537-41
[1] Isaacs, A., Lindenmann, J. Virus interference. 1. The interferon. Proc. R. Soc. Lond. B. Biol. Sci.1957 147:258–267.
[2] Velazquez,L., Fellous,M., Stark, G.R., Pellegrini, S. A protein tyrosine kinase in the interferon a/b signaling pathway. 1992 Cell 70: 313–322.
[3] Norimitsu Kadowaki ,Yong-Jun Liu. Natural Type I Interferon-Producing Cells as a Link Between Innate and Adaptive Immunity. Human Immunology 2002 63:1126–1132
[4] Andre Boonstra, Andrea M. Woltman. Immunology of hepatitis B and hepatitis C virus infections, Best Practice & Research Clinical Gastroenterology 2008 22:1049-1061
[5] Maria-Christina Jung ,Gerd R Pape. Immunology of hepatitis B infection. Lancet Infectious Diseases 2002 2:43-50
[6] Stefan Wieland,Robert Thimme. Genomic analysis of the host response to hepatitis B virus infection. PNAS 2004 101:6669-6674
[7] Kenya Honda1,Hideyuki Yanai,Hideo Negishi.IRF-7 is the master regulator of type-1 interferon-dependent immune responses. Nature 2005 434:772-777
[8] Patricia Fitzgerald-Bocarsly. Plasmacytoid dendritic cells and type I IFN:50 years of convergent history. Cytokine & Growth Factor Reviews 2008 19:3-19
[9] Renate G. van der Molen,Dave Sprengers. Functional Impairment of Myeloid and Plasmacytoid Dendritic Cells of Patients With Chronic Hepatitis B. Hepatology 2004 40:738-746
[10] Qing Xie,Huai-Cheng Shen,et al. Patients with chronic hepatitis B infection display deficiency of plasmacytoid dendritic cells with reduced expression of TLR9. Microbes and Infection. 2009 11:515-523
[11] Jian ZHOU, Yuancheng HUANG. Expression of Toll-like Receptor 9 in Peripheral Blood Mononuclear Cells from Patients with Different Hepatitis B and C Viral Loads. J Huazhong Univ Sci Technol. 2009 29(3):313-317
[12] Zheng Zhang, Hongfei Zhang.Response to interferon-αtreatment correlates with recovery of blood plasmacytoid dendritic cells in children with chronic hepatitis B. Journal of Hepatology 2007 47:751–759
[13] Luke AJ O’Neill. How Toll-like receptors signal: what we know and what we don’t know. Current Opinion in Immunology 2006, 18:3–9
[14] Shizuo Akira,Kiyoshi Takeda.Toll-like Receptor Signalling. Nature Reviews Immunology 2004 4:499-511
[15] Hemmi H,Takeuchi O, Kawai T, et al.A toll-like receptor recognizes bacterial DNA.Nature 2000 408:740-745
[16] Daniel B.Stetson, Ruslan Medzhitov. Type I Interferons in Host Defense.Immunity 2006 25:373-381
[17] Shizuo Akira, Satoshi Uematsu, Pathogen Recognition and Innate Immunity.Cell.2006 124:783-801
[18] Jennifer Lund, Ayuko Sato. Toll-like Receptor 9-mediated Recognition of Herpes Simplex Virus-2 by Plasmacytoid Dendritic Cells. J. Exp. Med 2003 198:513-520
[2] Lok, A.S, Heathcote, E.J, Hoofnagle, J.H. Management of hepatitis B: 2000-Summary of a workshop. Gastroenterology, 2001;120(7):1828-1853.
[3] Lau DT, Everhart J, Kleiner DE, Park Y, Vergalla J, Schmid P, Hoofnagle JH. Long-term follow-up of patients with chronic hepatitis B treated with interferon alfa. Gastroenterology 1997; 113: 1660-1667
[4] Patricia Fitzgerald-Bocarsly. Plasmacytoid dendritic cells and type I IFN:50 years of convergent history. Cytokine & Growth Factor Reviews 2008 19:3-19
[5] Shizuo Akira, Satoshi Uematsu, Pathogen Recognition and Innate Immunity. Cell.2006;124:783-801
[6]中华医学会肝病学分会,中华医学会感染病学分会.慢性乙型肝炎防治指南.2005
[7]黄呈辉,欧阳玲等,携带乙肝核心抗原基因复制缺陷型重组腺病毒的高效制备和表达。中国医学工程2006 ;14:616-621
[8] Stefan Bauer,Carsten J. Kirschning. Human TLR9 confers responsiveness to bacterial DNA via species-specific CpG motif recognition. PNAS .2001; 98(16) :9237–9242
[9] Ganem,D. and A.M. Prince.Hepatitis B virus infection-natural history and clinical consequences.N Engl J Med.2004;350(11):1118-1129
[10] Maria-Christina Jung ,Gerd R Pape. Immunology of hepatitis B infection. Lancet Infectious Diseases .2002 ;2:43-50
[11] Stefan Wieland,Robert Thimme. Genomic analysis of the host response to hepatitis B virus infection. PNAS.2004;101:6669-6674
[12] Luke AJ O’Neill. How Toll-like receptors signal: what we know and what we don’t know. Current Opinion in Immunology. 2006,;18:3–9
[13] Kenya Honda,Hideyuki Yanai.IRF-7 is the master regulator of type-I interferon-dependent immune responses. Nature .2005;434:772-777
[14] Norimitsu Kadowaki , Yong-Jun Liu. Natural Type I Interferon-Producing Cells as a Link Between Innate and Adaptive Immunity. Human Immunology.2002;63:1126–1132
[15] Kelli McKenna,Anne-Sophie Beignon. Plasmacytoid Dendritic Cells:Linking Innate and Adaptive Immunity.Journal of Virology.2005;79(1):17-27
[16] Gregory M. Barton,Jonathan C. Kagan. A cell biological view of Toll-like receptor function: regulation through compartmentalization. Nature Reviews Immunology. 2009;9:535-542
[17] Y Kumagai,O Takeuchi,S Akira. TLR 9 as a key receptor for the recognition of DNA. Advanced Drug Delivery Reviews.2008;60(7):795-804
[18] Jennifer Lund, Ayuko Sato. Toll-like Receptor 9-mediated Recognition of Herpes Simplex Virus-2 by Plasmacytoid Dendritic Cells. J. Exp. Med. 2003 ;198:513-520
[19] Jorg Schlender,Veit Hornung. Inhibition of toll-like receptor 7 and 9 mediated alpha/beta interferon production in human plasmacytoid dendritic cells by respiratory syncytial virus and measles virus. Journal of virology.2005;79(9):5507-5515
[20] Isogawa M. Robek MD.Furuich Y. Toll-like receptor signaling inhibits hepatitis B virus replication in vivo.J Virol.2005;79(11):7269-7272
[21] Fanning SL,George TC, Feng D, et al.Receptor cross-linking on human plasmacytoid dendritic cells leads to the regulation of IFN-alpha production.J Immunol 2006; 177:5829-39
[22] Feldman SB, Milone MC,Kloser P,Fitzgerald-Bocarsly P.Functional deficiencies in two distinct IFN-a producing cell populations in PBMC from human immunodeficiency virus seropositive patients.J Leu Biol.1995;57:214-20
[23] Heather W.Aging impairs IFN regulatory factor 7 up-regulation in plasmacytoid dendritic cells during TLR 9 activation. The Journal of Immunology.2008;181(10):6747-6756
[24] Chaperot L, Bendriss N, Manches O, Gressin R,et al. Identification of a leukemic counterpart of the plasmacytoid dendritic cells. Blood .2001;97:3210-7
[25] Heathcote J, Mchutchison J,Lee S,et al.A pilot study of C-1899 T-cell vaccine in subjects chronically infected with hepatitis B virus. The CY 1899 T-cell vaccine study group.Hepatology.1999;30:531-6
[26] Bertoletti A, Chisari FV, Penna A,et al. Definition of a minimal optimal cytotoxic T-cell epitope within the hepatitis B virus nucleocapsid protein. J Virol 1993;67:2376-80
[27] Ferrari C, Penna A,Bertoletti A,et al.Cellular immune response to hepatitis B virus-encoded antigens in acute and chronic hepatitis B virus infection.J Immunol 1990;145:3442-9
[28] Bertoletti A, Southwood S,Chesnut R,et al.Molecular features of the hepatitis B virus mucleocapsid T-cell epitopes 18-27:interaction with HLA and T-cell receptor. Hepatology.1997;26:1027-34
[29] Milich DR, Schodell F,Hughes JL,et al.The hepatitis B virus core and e antigens elicit different Th cell subsets: antigen structure can affect Th cell phenotype.J Virol 1997;71:2192-201
[30] Milich DR.Immune response to hepatitis B virus:infection,animal model,vaccine.Viral Hepat Rev 1997;3:63-103
[31] Qing Xie,Huai-Cheng Shen,et al. Patients with chronic hepatitis B infection displaydeficiency of plasmacytoid dendritic cells with reduced expression of TLR9. Microbes and Infection. 2009 11:515-523
[32] Jian ZHOU, Yuancheng HUANG. Expression of Toll-like Receptor 9 in Peripheral Blood Mononuclear Cells from Patients with Different Hepatitis B and C Viral Loads. J Huazhong Univ Sci Technol. 2009 29(3):313-317
[33] Wang FS, Xing LH, Liu MX, et al. Dysfunction of peripheral blood dendritic cells from patients with chronic hepatitis B virus infection. World J Gastroenterol 2001;7:537-41
[1] Isaacs, A., Lindenmann, J. Virus interference. 1. The interferon. Proc. R. Soc. Lond. B. Biol. Sci.1957 147:258–267.
[2] Velazquez,L., Fellous,M., Stark, G.R., Pellegrini, S. A protein tyrosine kinase in the interferon a/b signaling pathway. 1992 Cell 70: 313–322.
[3] Norimitsu Kadowaki ,Yong-Jun Liu. Natural Type I Interferon-Producing Cells as a Link Between Innate and Adaptive Immunity. Human Immunology 2002 63:1126–1132
[4] Andre Boonstra, Andrea M. Woltman. Immunology of hepatitis B and hepatitis C virus infections, Best Practice & Research Clinical Gastroenterology 2008 22:1049-1061
[5] Maria-Christina Jung ,Gerd R Pape. Immunology of hepatitis B infection. Lancet Infectious Diseases 2002 2:43-50
[6] Stefan Wieland,Robert Thimme. Genomic analysis of the host response to hepatitis B virus infection. PNAS 2004 101:6669-6674
[7] Kenya Honda1,Hideyuki Yanai,Hideo Negishi.IRF-7 is the master regulator of type-1 interferon-dependent immune responses. Nature 2005 434:772-777
[8] Patricia Fitzgerald-Bocarsly. Plasmacytoid dendritic cells and type I IFN:50 years of convergent history. Cytokine & Growth Factor Reviews 2008 19:3-19
[9] Renate G. van der Molen,Dave Sprengers. Functional Impairment of Myeloid and Plasmacytoid Dendritic Cells of Patients With Chronic Hepatitis B. Hepatology 2004 40:738-746
[10] Qing Xie,Huai-Cheng Shen,et al. Patients with chronic hepatitis B infection display deficiency of plasmacytoid dendritic cells with reduced expression of TLR9. Microbes and Infection. 2009 11:515-523
[11] Jian ZHOU, Yuancheng HUANG. Expression of Toll-like Receptor 9 in Peripheral Blood Mononuclear Cells from Patients with Different Hepatitis B and C Viral Loads. J Huazhong Univ Sci Technol. 2009 29(3):313-317
[12] Zheng Zhang, Hongfei Zhang.Response to interferon-αtreatment correlates with recovery of blood plasmacytoid dendritic cells in children with chronic hepatitis B. Journal of Hepatology 2007 47:751–759
[13] Luke AJ O’Neill. How Toll-like receptors signal: what we know and what we don’t know. Current Opinion in Immunology 2006, 18:3–9
[14] Shizuo Akira,Kiyoshi Takeda.Toll-like Receptor Signalling. Nature Reviews Immunology 2004 4:499-511
[15] Hemmi H,Takeuchi O, Kawai T, et al.A toll-like receptor recognizes bacterial DNA.Nature 2000 408:740-745
[16] Daniel B.Stetson, Ruslan Medzhitov. Type I Interferons in Host Defense.Immunity 2006 25:373-381
[17] Shizuo Akira, Satoshi Uematsu, Pathogen Recognition and Innate Immunity.Cell.2006 124:783-801
[18] Jennifer Lund, Ayuko Sato. Toll-like Receptor 9-mediated Recognition of Herpes Simplex Virus-2 by Plasmacytoid Dendritic Cells. J. Exp. Med 2003 198:513-520