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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 黃麗華 | |
dc.contributor.author | Yung-Chi Huang | en |
dc.contributor.author | 黃詠琪 | zh_TW |
dc.date.accessioned | 2021-06-13T01:35:26Z | - |
dc.date.available | 2009-08-08 | |
dc.date.copyright | 2007-08-08 | |
dc.date.issued | 2007 | |
dc.date.submitted | 2007-07-16 | |
dc.identifier.citation | 1. 2002. National Institutes of Health Consensus Development Conference Statement: Management of hepatitis C: 2002--June 10-12, 2002. Hepatology 36:S3-20.
2. Barba, G., F. Harper, T. Harada, M. Kohara, S. Goulinet, Y. Matsuura, G. Eder, Z. Schaff, M. J. Chapman, T. Miyamura, and C. Brechot. 1997. Hepatitis C virus core protein shows a cytoplasmic localization and associates to cellular lipid storage droplets. Proc Natl Acad Sci U S A 94:1200-5. 3. Bartenschlager, R., L. Ahlborn-Laake, J. Mous, and H. Jacobsen. 1994. Kinetic and structural analyses of hepatitis C virus polyprotein processing. J Virol 68:5045-55. 4. Bode, J. G., S. Ludwig, C. Ehrhardt, U. Albrecht, A. Erhardt, F. Schaper, P. C. Heinrich, and D. Haussinger. 2003. IFN-alpha antagonistic activity of HCV core protein involves induction of suppressor of cytokine signaling-3. Faseb J 17:488-90. 5. Bode, J. G., A. Nimmesgern, J. Schmitz, F. Schaper, M. Schmitt, W. Frisch, D. Haussinger, P. C. Heinrich, and L. Graeve. 1999. LPS and TNFalpha induce SOCS3 mRNA and inhibit IL-6-induced activation of STAT3 in macrophages. FEBS Lett 463:365-70. 6. Bonilla, P. J., S. A. Hughes, and S. R. Weiss. 1997. Characterization of a second cleavage site and demonstration of activity in trans by the papain-like proteinase of the murine coronavirus mouse hepatitis virus strain A59. J Virol 71:900-9. 7. Bressanelli, S., L. Tomei, F. A. Rey, and R. De Francesco. 2002. Structural analysis of the hepatitis C virus RNA polymerase in complex with ribonucleotides. J Virol 76:3482-92. 8. Bukh, J., R. H. Purcell, and R. H. Miller. 1992. Sequence analysis of the 5' noncoding region of hepatitis C virus. Proc Natl Acad Sci U S A 89:4942-6. 9. Butcher, S. J., J. M. Grimes, E. V. Makeyev, D. H. Bamford, and D. I. Stuart. 2001. A mechanism for initiating RNA-dependent RNA polymerization. Nature 410:235-40. 10. Caussin-Schwemling, C., C. Schmitt, and F. Stoll-Keller. 2001. Study of the infection of human blood derived monocyte/macrophages with hepatitis C virus in vitro. J Med Virol 65:14-22. 11. Chayama, K., A. Tsubota, M. Kobayashi, K. Okamoto, M. Hashimoto, Y. Miyano, H. Koike, M. Kobayashi, I. Koida, Y. Arase, S. Saitoh, Y. Suzuki, N. Murashima, K. Ikeda, and H. Kumada. 1997. Pretreatment virus load and multiple amino acid substitutions in the interferon sensitivity-determining region predict the outcome of interferon treatment in patients with chronic genotype 1b hepatitis C virus infection. Hepatology 25:745-9. 12. Choo, Q. L., G. Kuo, A. J. Weiner, L. R. Overby, D. W. Bradley, and M. Houghton. 1989. Isolation of a cDNA clone derived from a blood-borne non-A, non-B viral hepatitis genome. Science 244:359-62. 13. Choo, Q. L., K. H. Richman, J. H. Han, K. Berger, C. Lee, C. Dong, C. Gallegos, D. Coit, R. Medina-Selby, P. J. Barr, and et al. 1991. Genetic organization and diversity of the hepatitis C virus. Proc Natl Acad Sci U S A 88:2451-5. 14. Coito, C., D. L. Diamond, P. Neddermann, M. J. Korth, and M. G. Katze. 2004. High-throughput screening of the yeast kinome: identification of human serine/threonine protein kinases that phosphorylate the hepatitis C virus NS5A protein. J Virol 78:3502-13. 15. Demaison, C., K. Parsley, G. Brouns, M. Scherr, K. Battmer, C. Kinnon, M. Grez, and A. J. Thrasher. 2002. High-level transduction and gene expression in hematopoietic repopulating cells using a human immunodeficiency [correction of imunodeficiency] virus type 1-based lentiviral vector containing an internal spleen focus forming virus promoter. Hum Gene Ther 13:803-13. 16. Dolganiuc, A., S. Chang, K. Kodys, P. Mandrekar, G. Bakis, M. Cormier, and G. Szabo. 2006. Hepatitis C virus (HCV) core protein-induced, monocyte-mediated mechanisms of reduced IFN-alpha and plasmacytoid dendritic cell loss in chronic HCV infection. J Immunol 177:6758-68. 17. Dolganiuc, A., S. Oak, K. Kodys, D. T. Golenbock, R. W. Finberg, E. Kurt-Jones, and G. Szabo. 2004. Hepatitis C core and nonstructural 3 proteins trigger toll-like receptor 2-mediated pathways and inflammatory activation. Gastroenterology 127:1513-24. 18. Duong, F. H., M. Filipowicz, M. Tripodi, N. La Monica, and M. H. Heim. 2004. Hepatitis C virus inhibits interferon signaling through up-regulation of protein phosphatase 2A. Gastroenterology 126:263-77. 19. Egger, D., B. Wolk, R. Gosert, L. Bianchi, H. E. Blum, D. Moradpour, and K. Bienz. 2002. Expression of hepatitis C virus proteins induces distinct membrane alterations including a candidate viral replication complex. J Virol 76:5974-84. 20. Failla, C., L. Tomei, and R. De Francesco. 1994. Both NS3 and NS4A are required for proteolytic processing of hepatitis C virus nonstructural proteins. J Virol 68:3753-60. 21. Foy, E., K. Li, C. Wang, R. Sumpter, Jr., M. Ikeda, S. M. Lemon, and M. Gale, Jr. 2003. Regulation of interferon regulatory factor-3 by the hepatitis C virus serine protease. Science 300:1145-8. 22. Fried, M. W., M. L. Shiffman, K. R. Reddy, C. Smith, G. Marinos, F. L. Goncales, Jr., D. Haussinger, M. Diago, G. Carosi, D. Dhumeaux, A. Craxi, A. Lin, J. Hoffman, and J. Yu. 2002. Peginterferon alfa-2a plus ribavirin for chronic hepatitis C virus infection. N Engl J Med 347:975-82. 23. Garcia-Sastre, A., and C. A. Biron. 2006. Type 1 interferons and the virus-host relationship: a lesson in detente. Science 312:879-82. 24. Ghosh, A. K., M. Majumder, R. Steele, K. Meyer, R. Ray, and R. B. Ray. 2000. Hepatitis C virus NS5A protein protects against TNF-alpha mediated apoptotic cell death. Virus Res 67:173-8. 25. Gretch, D. R. 1997. Diagnostic tests for hepatitis C. Hepatology 26:43S-47S. 26. Gutterman, J. U. 1994. Cytokine therapeutics: lessons from interferon alpha. Proc Natl Acad Sci U S A 91:1198-205. 27. Hadziyannis, S. J., H. Sette, Jr., T. R. Morgan, V. Balan, M. Diago, P. Marcellin, G. Ramadori, H. Bodenheimer, Jr., D. Bernstein, M. Rizzetto, S. Zeuzem, P. J. Pockros, A. Lin, and A. M. Ackrill. 2004. Peginterferon-alpha2a and ribavirin combination therapy in chronic hepatitis C: a randomized study of treatment duration and ribavirin dose. Ann Intern Med 140:346-55. 28. Hassan, M., D. Selimovic, H. Ghozlan, and O. Abdel-Kader. 2007. Induction of high-molecular-weight (HMW) tumor necrosis factor(TNF) alpha by hepatitis C virus (HCV) non-structural protein 3 (NS3) in liver cells is AP-1 and NF-kappaB-dependent activation. Cell Signal 19:301-11. 29. Hayashi, N., and T. Takehara. 2006. Antiviral therapy for chronic hepatitis C: past, present, and future. J Gastroenterol 41:17-27. 30. Herbein, G., and W. A. O'Brien. 2000. Tumor necrosis factor (TNF)-alpha and TNF receptors in viral pathogenesis. Proc Soc Exp Biol Med 223:241-57. 31. Hijikata, M., H. Mizushima, Y. Tanji, Y. Komoda, Y. Hirowatari, T. Akagi, N. Kato, K. Kimura, and K. Shimotohno. 1993. Proteolytic processing and membrane association of putative nonstructural proteins of hepatitis C virus. Proc Natl Acad Sci U S A 90:10773-7. 32. Hoofnagle, J. H., and L. B. Seeff. 2006. Peginterferon and ribavirin for chronic hepatitis C. N Engl J Med 355:2444-51. 33. Houghton, M., and S. Abrignani. 2005. Prospects for a vaccine against the hepatitis C virus. Nature 436:961-6. 34. Ide, Y., A. Tanimoto, Y. Sasaguri, and R. Padmanabhan. 1997. Hepatitis C virus NS5A protein is phosphorylated in vitro by a stably bound protein kinase from HeLa cells and by cAMP-dependent protein kinase A-alpha catalytic subunit. Gene 201:151-8. 35. Ito, T., S. M. Tahara, and M. M. Lai. 1998. The 3'-untranslated region of hepatitis C virus RNA enhances translation from an internal ribosomal entry site. J Virol 72:8789-96. 36. Kadoya, H., M. Nagano-Fujii, L. Deng, N. Nakazono, and H. Hotta. 2005. Nonstructural proteins 4A and 4B of hepatitis C virus transactivate the interleukin 8 promoter. Microbiol Immunol 49:265-73. 37. Kato, N., Y. Ootsuyama, S. Ohkoshi, T. Nakazawa, H. Sekiya, M. Hijikata, and K. Shimotohno. 1992. Characterization of hypervariable regions in the putative envelope protein of hepatitis C virus. Biochem Biophys Res Commun 189:119-27. 38. Katze, M. G., Y. He, and M. Gale, Jr. 2002. Viruses and interferon: a fight for supremacy. Nat Rev Immunol 2:675-87. 39. Khabar, K. S., F. Al-Zoghaibi, M. N. Al-Ahdal, T. Murayama, M. Dhalla, N. Mukaida, M. Taha, S. T. Al-Sedairy, Y. Siddiqui, G. Kessie, and K. Matsushima. 1997. The alpha chemokine, interleukin 8, inhibits the antiviral action of interferon alpha. J Exp Med 186:1077-85. 40. Kim, D. W., R. Suzuki, T. Harada, I. Saito, and T. Miyamura. 1994. Trans-suppression of gene expression by hepatitis C viral core protein. Jpn J Med Sci Biol 47:211-20. 41. Lam, A. M., D. Keeney, P. Q. Eckert, and D. N. Frick. 2003. Hepatitis C virus NS3 ATPases/helicases from different genotypes exhibit variations in enzymatic properties. J Virol 77:3950-61. 42. Lane, B. R., K. Lore, P. J. Bock, J. Andersson, M. J. Coffey, R. M. Strieter, and D. M. Markovitz. 2001. Interleukin-8 stimulates human immunodeficiency virus type 1 replication and is a potential new target for antiretroviral therapy. J Virol 75:8195-202. 43. Lee, J. C., Y. F. Shih, S. P. Hsu, T. Y. Chang, L. H. Chen, and J. T. Hsu. 2003. Development of a cell-based assay for monitoring specific hepatitis C virus NS3/4A protease activity in mammalian cells. Anal Biochem 316:162-70. 44. Li, K., Z. Chen, N. Kato, M. Gale, Jr., and S. M. Lemon. 2005. Distinct poly(I-C) and virus-activated signaling pathways leading to interferon-beta production in hepatocytes. J Biol Chem 280:16739-47. 45. Li, K., E. Foy, J. C. Ferreon, M. Nakamura, A. C. Ferreon, M. Ikeda, S. C. Ray, M. Gale, Jr., and S. M. Lemon. 2005. Immune evasion by hepatitis C virus NS3/4A protease-mediated cleavage of the Toll-like receptor 3 adaptor protein TRIF. Proc Natl Acad Sci U S A 102:2992-7. 46. Li, X. D., L. Sun, R. B. Seth, G. Pineda, and Z. J. Chen. 2005. Hepatitis C virus protease NS3/4A cleaves mitochondrial antiviral signaling protein off the mitochondria to evade innate immunity. Proc Natl Acad Sci U S A 102:17717-22. 47. Liu, B., J. Liao, X. Rao, S. A. Kushner, C. D. Chung, D. D. Chang, and K. Shuai. 1998. Inhibition of Stat1-mediated gene activation by PIAS1. Proc Natl Acad Sci U S A 95:10626-31. 48. Liu, Q., C. Tackney, R. A. Bhat, A. M. Prince, and P. Zhang. 1997. Regulated processing of hepatitis C virus core protein is linked to subcellular localization. J Virol 71:657-62. 49. Loo, Y. M., D. M. Owen, K. Li, A. K. Erickson, C. L. Johnson, P. M. Fish, D. S. Carney, T. Wang, H. Ishida, M. Yoneyama, T. Fujita, T. Saito, W. M. Lee, C. H. Hagedorn, D. T. Lau, S. A. Weinman, S. M. Lemon, and M. Gale, Jr. 2006. Viral and therapeutic control of IFN-beta promoter stimulator 1 during hepatitis C virus infection. Proc Natl Acad Sci U S A 103:6001-6. 50. Makarova, O., E. Kamberov, and B. Margolis. 2000. Generation of deletion and point mutations with one primer in a single cloning step. Biotechniques 29:970-2. 51. Manns, M. P., J. G. McHutchison, S. C. Gordon, V. K. Rustgi, M. Shiffman, R. Reindollar, Z. D. Goodman, K. Koury, M. Ling, and J. K. Albrecht. 2001. Peginterferon alfa-2b plus ribavirin compared with interferon alfa-2b plus ribavirin for initial treatment of chronic hepatitis C: a randomised trial. Lancet 358:958-65. 52. Mbow, M. L., and R. T. Sarisky. 2004. What is disrupting IFN-alpha's antiviral activity? Trends Biotechnol 22:395-9. 53. Meylan, E., J. Curran, K. Hofmann, D. Moradpour, M. Binder, R. Bartenschlager, and J. Tschopp. 2005. Cardif is an adaptor protein in the RIG-I antiviral pathway and is targeted by hepatitis C virus. Nature 437:1167-72. 54. Miyamura, T., and Y. Matsuura. 1993. Structural proteins of hepatitis C virus. Trends Microbiol 1:229-31. 55. Murayama, T., K. Kuno, F. Jisaki, M. Obuchi, D. Sakamuro, T. Furukawa, N. Mukaida, and K. Matsushima. 1994. Enhancement human cytomegalovirus replication in a human lung fibroblast cell line by interleukin-8. J Virol 68:7582-5. 56. Neuman, M. G., J. P. Benhamou, I. M. Malkiewicz, R. Akremi, N. H. Shear, T. Asselah, A. Ibrahim, N. Boyer, M. Martinot-Peignoux, P. Jacobson-Brown, G. G. Katz, V. Le Breton, G. Le Guludec, A. Suneja, and P. Marcellin. 2001. Cytokines as predictors for sustained response and as markers for immunomodulation in patients with chronic hepatitis C. Clin Biochem 34:173-82. 57. Noguchi, T., S. Satoh, T. Noshi, E. Hatada, R. Fukuda, A. Kawai, S. Ikeda, M. Hijikata, and K. Shimotohno. 2001. Effects of mutation in hepatitis C virus nonstructural protein 5A on interferon resistance mediated by inhibition of PKR kinase activity in mammalian cells. Microbiol Immunol 45:829-40. 58. Pavlovic, D., D. C. Neville, O. Argaud, B. Blumberg, R. A. Dwek, W. B. Fischer, and N. Zitzmann. 2003. The hepatitis C virus p7 protein forms an ion channel that is inhibited by long-alkyl-chain iminosugar derivatives. Proc Natl Acad Sci U S A 100:6104-8. 59. Pawlotsky, J. M. 2004. Pathophysiology of hepatitis C virus infection and related liver disease. Trends Microbiol 12:96-102. 60. Polyak, S. J., K. S. Khabar, D. M. Paschal, H. J. Ezelle, G. Duverlie, G. N. Barber, D. E. Levy, N. Mukaida, and D. R. Gretch. 2001. Hepatitis C virus nonstructural 5A protein induces interleukin-8, leading to partial inhibition of the interferon-induced antiviral response. J Virol 75:6095-106. 61. Polyak, S. J., K. S. Khabar, M. Rezeiq, and D. R. Gretch. 2001. Elevated levels of interleukin-8 in serum are associated with hepatitis C virus infection and resistance to interferon therapy. J Virol 75:6209-11. 62. Radkowski, M., A. Bednarska, A. Horban, J. Stanczak, J. Wilkinson, D. M. Adair, M. Nowicki, J. Rakela, and T. Laskus. 2004. Infection of primary human macrophages with hepatitis C virus in vitro: induction of tumour necrosis factor-alpha and interleukin 8. J Gen Virol 85:47-59. 63. Ray, R. B., K. Meyer, and R. Ray. 1996. Suppression of apoptotic cell death by hepatitis C virus core protein. Virology 226:176-82. 64. Reed, K. E., J. Xu, and C. M. Rice. 1997. Phosphorylation of the hepatitis C virus NS5A protein in vitro and in vivo: properties of the NS5A-associated kinase. J Virol 71:7187-97. 65. Santolini, E., L. Pacini, C. Fipaldini, G. Migliaccio, and N. Monica. 1995. The NS2 protein of hepatitis C virus is a transmembrane polypeptide. J Virol 69:7461-71. 66. Seth, R. B., L. Sun, C. K. Ea, and Z. J. Chen. 2005. Identification and characterization of MAVS, a mitochondrial antiviral signaling protein that activates NF-kappaB and IRF 3. Cell 122:669-82. 67. Shi, S. T., and M. M. Lai. 2001. Hepatitis C viral RNA: challenges and promises. Cell Mol Life Sci 58:1276-95. 68. Shih, C. M., S. J. Lo, T. Miyamura, S. Y. Chen, and Y. H. Lee. 1993. Suppression of hepatitis B virus expression and replication by hepatitis C virus core protein in HuH-7 cells. J Virol 67:5823-32. 69. Simmonds, P. 2004. Genetic diversity and evolution of hepatitis C virus--15 years on. J Gen Virol 85:3173-88. 70. Simoncic, P. D., A. Lee-Loy, D. L. Barber, M. L. Tremblay, and C. J. McGlade. 2002. The T cell protein tyrosine phosphatase is a negative regulator of janus family kinases 1 and 3. Curr Biol 12:446-53. 71. Sumpter, R., Jr., Y. M. Loo, E. Foy, K. Li, M. Yoneyama, T. Fujita, S. M. Lemon, and M. Gale, Jr. 2005. Regulating intracellular antiviral defense and permissiveness to hepatitis C virus RNA replication through a cellular RNA helicase, RIG-I. J Virol 79:2689-99. 72. Tai, C. L., W. K. Chi, D. S. Chen, and L. H. Hwang. 1996. The helicase activity associated with hepatitis C virus nonstructural protein 3 (NS3). J Virol 70:8477-84. 73. Tanji, Y., M. Hijikata, S. Satoh, T. Kaneko, and K. Shimotohno. 1995. Hepatitis C virus-encoded nonstructural protein NS4A has versatile functions in viral protein processing. J Virol 69:1575-81. 74. Taylor, D. R., S. T. Shi, P. R. Romano, G. N. Barber, and M. M. Lai. 1999. Inhibition of the interferon-inducible protein kinase PKR by HCV E2 protein. Science 285:107-10. 75. Tong, X., Z. Guo, J. Wright-Minogue, E. Xia, A. Prongay, V. Madison, P. Qiu, S. Venkatraman, F. Velazquez, F. G. Njoroge, and B. A. Malcolm. 2006. Impact of naturally occurring variants of HCV protease on the binding of different classes of protease inhibitors. Biochemistry 45:1353-61. 76. Vlotides, G., A. S. Sorensen, F. Kopp, K. Zitzmann, N. Cengic, S. Brand, R. Zachoval, and C. J. Auernhammer. 2004. SOCS-1 and SOCS-3 inhibit IFN-alpha-induced expression of the antiviral proteins 2,5-OAS and MxA. Biochem Biophys Res Commun 320:1007-14. 77. Wang, C., P. Sarnow, and A. Siddiqui. 1993. Translation of human hepatitis C virus RNA in cultured cells is mediated by an internal ribosome-binding mechanism. J Virol 67:3338-44. 78. Wolk, B., D. Sansonno, H. G. Krausslich, F. Dammacco, C. M. Rice, H. E. Blum, and D. Moradpour. 2000. Subcellular localization, stability, and trans-cleavage competence of the hepatitis C virus NS3-NS4A complex expressed in tetracycline-regulated cell lines. J Virol 74:2293-304. 79. Yoshimura, A., H. Nishinakamura, Y. Matsumura, and T. Hanada. 2005. Negative regulation of cytokine signaling and immune responses by SOCS proteins. Arthritis Res Ther 7:100-10. 80. Zein, N. N. 2000. Clinical significance of hepatitis C virus genotypes. Clin Microbiol Rev 13:223-35. 81. Zhu, N., A. Khoshnan, R. Schneider, M. Matsumoto, G. Dennert, C. Ware, and M. M. Lai. 1998. Hepatitis C virus core protein binds to the cytoplasmic domain of tumor necrosis factor (TNF) receptor 1 and enhances TNF-induced apoptosis. J Virol 72:3691-7. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/30089 | - |
dc.description.abstract | C型肝炎病毒 (HCV)感染無論在台灣或是全球都是很重要的公共衛生課題。根據世界衛生組織的統計,全世界約有一億七千萬的人口為HCV帶原者,其中高達百分之八十的人在二十到三十年內會產生肝硬化,甚且發展成惡性肝細胞癌。目前最有效的治療藥物為長效型甲型干擾素結合抗病毒藥物ribavirin,然而,隨著病毒基因型的不同,治療成效約在50-80%不等。同時,感染後發展成慢性肝炎的機率以及所伴隨的肝臟疾病的嚴重程度在病毒基因型間也有所不同。本研究主要想澄清基因型1b和2a病毒 (台灣最普遍的兩種基因型)的NS3/4A對於細胞中干擾素的產生及治療作用是否有所不同,最後導致臨床上所觀察到的差異。我們從病人血清中分離出分屬基因型1b和2a各三個NS3/4A clones。首先,檢視其對於IFN-β啟動子驅使的冷光素酵素報導基因之活性的抑制及對MAVS的切割能力。並且,利用ISRE啟動子驅使的冷光素酵素報導基因之活性和trans-rescue測定,觀察NS3/4A對IFN-α下游訊息傳導抑制的能力。根據以上的分析結果,並未發現基因型1b和2a的NS3/4A有能力上的差異。因此,推測C型肝炎病毒中的NS3/4A對於不同基因型病毒感染的患者所表現出的臨床差異上,並未扮演關鍵性的角色。論文中的第二部分,我們試圖找出能加強細胞對干擾素負向調控的C型肝炎病毒蛋白質。根據初步的結果,發現在肝癌細胞中表現C型肝炎病毒的core會增加細胞中SOCS3 mRNA的量。另外,表現NS3/4A,,NS4B 和NS5A 則能使人類單核球細胞中抑制細胞抗病毒作用的細胞激素IL-8 與TNF-α的表現量增加。對於C型肝炎病毒如何負向調控細胞中干擾素的作用,並且增加對IFN-α治療的抵抗,則需更深入的研究。 | zh_TW |
dc.description.abstract | Hepatitis C virus (HCV) is a major etiologic pathogen that infects 170 million people worldwide and causes chronic hepatitis, which often leads to liver cirrhosis, and hepatocellular carcinoma. The standard therapy for chronic HCV infection is a combination of PEG-IFN-α (pegylated-interferon-α) and ribavirin, which could eliminate HCV in 50-80% of the patients with chronic active hepatitis. However, the rate of successful elimination varies significantly with virus genotypes. Also, the rate of evolution to chronic hepatitis and the severity of liver disease are different among the patients infected by different HCV genotypes. Here, we attempted to clarify whether the different clinical outcomes in patients infected with HCV genotypes 1b and 2a, the two most prevalent genotypes in Taiwan, were due to the different inhibitory effects of the NS3/4A proteins on cellular IFN response. Three clones of each HCV genotype 1b or 2a were isolated from patients. We first compared the efficacies of these NS3/4A in interrupting the IFN-β (interferon-β) promoter activation and cleaving the MAVS (mitochondrial antiviral signaling) protein; and secondly, we evaluated their abilities to disrupt the IFN-α-induced signaling by means of ISRE (interferon stimulated response element) activity and the virus trans-rescue assay. For all the analyses, we found that there’s no significant differences between NS3/4A from genotypes 1b and 2a, suggesting that HCV NS3/4A might not play a critical role in the different clinical outcomes of the patients infected by the two studied HCV genotypes. In the second part of this study, we screened for the possible HCV proteins which could augment the negative regulation of IFN signaling. Our preliminary data showed that expression of HCV core and NS4B in a hepatoma cell line increased the SOCS3 mRNA levels, and the expression of NS3/4A, NS4B and NS5A in human monocyte cell line enhanced the expression of cytokine IL-8 and TNF-α, which are all known able to inhibit cellular antiviral activity. The underlying mechanisms for the augmented negative regulation of IFN induced by HCV proteins remain to be elucidated. | en |
dc.description.provenance | Made available in DSpace on 2021-06-13T01:35:26Z (GMT). No. of bitstreams: 1 ntu-96-R94445118-1.pdf: 11485512 bytes, checksum: 3c7b45b067c63be0d461e6e675897528 (MD5) Previous issue date: 2007 | en |
dc.description.tableofcontents | 中文摘要 i
英文摘要 ii 縮寫對照表 iv 目錄 vi 壹、緒論 1 一、C型肝炎病毒 (Hepatitis C virus) 1 1. C型肝炎病毒基因體結構 1 2. C型肝炎病毒各基因體產物 2 二、C型肝炎病毒流行病學 5 1. C型肝炎病毒的傳染 5 2. C型肝炎病毒感染的治療 6 3. C型肝炎病毒的變異及其臨床上意義 7 三、C型肝炎病毒與宿主細胞抗病毒防禦機制的交互作用 8 1. 病毒感染後宿主的防禦反應 8 2. 細胞中負向調控干擾素作用的機制 9 3. C型肝炎病毒拮抗細胞所產生的干擾素反應 9 四、研究方向及目的 11 貳、材料與方法 13 一、化學藥劑與抗體 13 二、細胞株培養與病毒感染 13 三、從C型肝炎患者血清中cloning出NS3/4A所使用之引子 14 四、NS3/4A蛋白酶突變株S139A的構築 14 五、試管內 (in vitro)轉錄及轉譯所進行之分析 15 六、DNA轉染 16 七、冷光素酵素報導基因分析 (luciferase reporter assay) 16 八、VSV trans-rescue及plaque assay 17 九、以Lentivirus作為病毒載體攜帶基因到單核球細胞中表現 18 十、西方點墨法 18 十一、同步定量聚合連鎖反應 (quantitative Real-time RT-PCR) 19 參、結果 21 一、不同基因型的C型肝炎病毒NS3/4A對細胞宿主干擾素的產生及 作用之影響 21 1. 從慢性C型肝炎患者血清中cloning出非結構性蛋白NS3/4A及 蛋白酶突變株質體的構築 21 2. 在恆定狀態 (steady state)下觀察NS3/4A對細胞中MAVS蛋白質 的切割能力及IFN-β啟動子驅使的冷光素酵素活性的抑制情形 23 3. 以外送的MAVS觀察NS3/4A對MAVS蛋白質的切割能力及其 對IFN-β啟動子驅使的冷光素酵素活性的抑制情形 24 4. NS3/4A對人工受質EGFP-(△MAVS)-SEAP的切割能力 25 5. 在in vitro中分析NS3/4A形成功能完整 (functional)的蛋白酶複合 體之速率 27 6. NS3/4A對ISRE驅使的冷光素酵素活性的抑制情形 28 7. 利用VSV病毒trans-rescue分析方法來觀察NS3/4A拮抗IFN-α 的能力 28 二、 C型肝炎病毒增強細胞對干擾素訊息傳導的負向調控 29 1. C型肝炎病毒蛋白質對於肝癌細胞中SOCS3表現量的影響 29 2. C型肝炎病毒蛋白質對單核球細胞中細胞激素IL-8及TNF-α表現量 的影響 30 肆、討論 31 一、不同基因型的NS3/4A蛋白質對於臨床病人本身干擾素產生,以及對 干擾素治療的反應之影響 31 二、C型肝炎病毒增強細胞對干擾素下游訊息傳導的負向調控 34 伍、附圖 38 陸、參考文獻 59 | |
dc.language.iso | zh-TW | |
dc.title | C型肝炎病毒基因型1b和2a非結構性蛋白質NS3/4A對細胞干擾素反應的抑制能力之比較 | zh_TW |
dc.title | The comparison of different inhibitory effects of
hepatitis C virus NS3/4A from genotypes 1b and 2a on cellular interferon responses | en |
dc.type | Thesis | |
dc.date.schoolyear | 95-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 林宜玲,陳美如,董馨蓮 | |
dc.subject.keyword | C型肝炎病毒,干擾素,病毒基因型, | zh_TW |
dc.subject.keyword | Hepatitis C virus,interferon,HCV genotypes, | en |
dc.relation.page | 66 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2007-07-16 | |
dc.contributor.author-college | 醫學院 | zh_TW |
dc.contributor.author-dept | 微生物學研究所 | zh_TW |
顯示於系所單位: | 微生物學科所 |
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