進階搜尋


 
系統識別號 U0026-0812200910454278
論文名稱(中文) 抗登革病毒非結構性蛋白1抗體對於肝臟損害及內皮細胞活化之病理效應
論文名稱(英文) Pathological effects of antibodies against dengue virus nonstructural protein 1 in liver damage and endothelial cell activation
校院名稱 成功大學
系所名稱(中) 微生物及免疫學研究所
系所名稱(英) Department of Microbiology & Immunology
學年度 91
學期 2
出版年 92
研究生(中文) 蕭玉翎
研究生(英文) Yu-Ling Hsiao
學號 s4690408
學位類別 碩士
語文別 中文
論文頁數 74頁
口試委員 口試委員-張德卿
指導教授-林以行
口試委員-黎煥耀
口試委員-劉清泉
中文關鍵字 非結構性蛋白1  登革病毒 
英文關鍵字 dengue virus  NS1 
學科別分類
中文摘要 登革病毒感染往往導致輕微的登革熱或者威脅生命的登革出血熱/登革休克症狀。然而,對於登革感染的致病機轉至今仍無完全的瞭解。在本實驗室先前的研究中發現,無論是在主動模式下給予小鼠登革病毒非結構性蛋白1 (DV NS1) 或是被動給予小鼠抗DV NS1抗體,都發現小鼠肝臟中的AST/ALT濃度有增高的情形,但其腎臟之BUN值並無顯著的變化。另外也發現抗DV NS1抗體可與小鼠的下腔靜脈結合。在本論文中,我們進一步探究抗DV NS1抗體在受損的器官中可能扮演的角色。結果發現在主動給予DV NS1小鼠的肝臟血管內皮層有抗體的堆積且伴隨著有細胞凋亡的情形,但在腎臟並無觀察到有特異性抗體的堆積以及細胞凋亡的現象。另一方面,我們也發現抗DV NS1抗體可直接與正常小鼠肝臟血管內皮層結合,但在腎臟並沒有發現特異性的結合情形。除此之外,主動給予DV NS1的小鼠肝臟中有巨噬細胞浸潤的情形。我們推測因DV NS1抗體刺激而造成的內皮細胞功能喪失可能與肝臟中浸潤的細胞一同造成肝臟的損害。先前本實驗室在體外實驗中觀察到在抗DV NS1抗體刺激內皮細胞後會造成免疫活化作用,包括細胞激素、化學趨化激素以及細胞黏著分子的表現量上升。在本論文中,我們發現這些物質的活化與NF-B相關,藉由抑制其活性可降低IL-6、IL-8、MCP-1及部分ICAM-1的表現。綜合以上體內及體外的研究結果,對於抗DV NS1抗體刺激所造成的自體免疫作用,並導致免疫活化現象,可能顯示抗DV NS1抗體在登革感染的致病機轉中扮演了一個重要的角色。
英文摘要 Dengue virus (DV) infection causes dengue fever or severe life-threatening dengue hemorrhagic fever/dengue shock syndrome (DHF/DSS). However, the pathogenesis of DHF/DSS is still not well understood. Previous studies in our laboratory showed that the AST and ALT, but not BUN, in mouse sera increased after either active immunization with DV nonstructural protein 1 (NS1) or passive administration with anti-DV NS1 antibodies (Abs). Furthermore, anti-DV NS1 Abs could bind to mouse vessel endothelium in inferior vena. In this study, we investigate the potential pathogenic role of anti-DV NS1 Abs in the damaged organs. Mice actively immunized with DV NS1 protein revealed the presence of anti-DV NS1 Abs on vessel endothelium in the liver. However, in the kidney, mice actively immunized with DV NS1 protein, JEV NS1 protein or PBS show similar binding patterns. Studies using organ sections from normal mice showed that anti-DV NS1 Abs, but not anti-JEV NS1 Abs, could directly bind to vessel endothelium in the liver sections. However, no specific cross-reaction of anti-DV NS1 Abs with mouse kidney vessel endothelium. Interestingly, gross and histological examinations revealed tissue damage with the presence of macrophage infiltration in the liver of NS1-immunized mice. We speculate that anti-DV NS1-induced endothelial cell dysfunction might result in liver tissue damage, at least in part, by infiltrated cells. Previous in vitro studies in our laboratory showed immune activation including cytokine and chemokine production in endothelial cells after treatment with anti-DV NS1 Abs. In this study, we demonstrated the involvement of upstream regulator NF-kB in immune activation by EMSA. After treatment with NF-kB inhibitor pyrrolidine dithiocarbamate, the expressions of IL-6、IL-8 and MCP-1 which could be stimulated by anti-DV NS1 Abs were inhibited. Taken together, endothelial cell damage and abnormal activation after stimulation by anti-DV NS1 Abs have been examined both in vivo and in vitro. These results imply the involvement of anti-DV NS1 Abs in the pathogenesis of DV infection.
論文目次 中文摘要I
英文摘要II
致謝III
目錄IV
圖目錄VII
符號及縮寫索引表IX

緒論1
材料與方法7
A. 材料
A-1 實驗動物 7
A-2 細菌株 7
A-3 細胞株 7
A-4 試劑藥品8
A-5 抗體 10
A-6 耗材 12
A-7 儀器 13

B. 方法
B-1 NS1重組蛋白之製備 15
B-1.1 NS1重組蛋白之誘導表現 15
B-1.2 NS1重組蛋白的純化 15

B-2 SDS-PAGE 膠體電泳 16
B-2.1 SDS-PAGE之蛋白質電泳步驟 16
B-2.2以細菌為樣本時的處理17
B-3 蛋白質定量17
B-4 NS1重組蛋白免疫小鼠及血清的製備18
B-5 抗體的製備與特性分析18
B-5.1 抗體的製備18
B-5.2 抗體特性的分析-以酵素結合免疫吸附法 (Enzyme-linked immunosorbent assay, ELISA) 測定抗體的效價19
B-6 rNS1免疫小鼠病理現象之觀察19
B-6.1 石蠟組織切片的製備與HE stain 20
B-6.2 冷凍切片的製備20
B-6.3 以螢光染色觀察器官的病理變化20
B-7 以免疫組織化學染色法(Immunohistochemistry, IHC) 觀察細胞死亡的情形20
B-7.1 玻片的前處理與組織切片的製備請參考B-6.1及B-6.2 21
B-7.2 組織切片的TUNEL反應21
B-8 以電泳移動偏移分析( Electrophoretic Mobility Shift Assays,EMSA ) 探討核蛋白之活化22
B-8.1細胞核內蛋白質萃取22
B-8.2 EMSA 的操作23
B-9以免疫細胞染色 (immunocytochemistrys taining) 法觀察細胞激素、化學趨化激素以及細胞黏著分子的表現24
B-9.1 Cytokine、chemokine以及adhesion molecule之細胞染色24


結果
I. 探討登革病毒非結構性蛋白1抗體與血管內皮層發生交叉結合作用25
1. 抗登革病毒非結構性蛋白1 (DV NS1) 抗體致病性角色的研究25
2.以DV NS1重組蛋白免疫小鼠建立主動模式研究抗體致病性26
3.主動模式中觀察抗DV NS1抗體造成肝臟及腎臟之病理現象27
II. 探討抗登革病毒非結構性蛋白1抗體刺激內皮細胞發生訊息傳遞作用29
1. 抗DV NS1抗體刺激內皮細胞造成NF-kB的活化30
2.探討NF-kB在抗登革病毒非結構性蛋白1抗體活化內皮細胞過程中所扮演的角色30

III. 抗登革病毒非結構性蛋白1抗體造成內皮系統異常活化與肝臟病變的關係32

討論34
參考文獻40
圖50
附錄68
作者簡歷74
參考文獻 Avirutnan, P., P. Malasit, B. Seliger, S. Bhakdi, and M. Husmann. 1998. Dengue virus infection of human endothelial cells leads to chemokine production, complement activation, and apoptosis. J. Immunol. 161:6338.

Baldwin A. S. 1996. The NFkB and IkB proteins: new discoveries and insights. Annu. Rev. Immunol. 14:649.

Bessho, R., K. Matsubara, M. Kubota, K. Kuwakado, H. Hirota, Y. Wakazono, Y. W. Lin, A. Okuda, M. Kawai, and R. Nishikomori. 1994. Pyrrolidine dithiocarbamate, a potent inhibitor of nuclear factor kappa B (NF-kappa B) activation, prevents apoptosis in human promyelocytic leukemia HL-60 cells and thymocytes. Biochem. Pharmacol. 48:1883.

Bhakdi, S., and M. D. Kazatchkine. 1990. Pathogenesis of dengue: an alternative hypothesis. Southeast Asian J. Trop. Med. Public Health 21:652.

Bhamarapravati, N. 1989. Hemostatic defects in dengue hemorrhagic fever. Rev. Infect. Dis. 11:S826.

Bielefeldt-Ohmann, H. 1997. Pathogenesis of dengue virus diseases: missing pieces in the jigsaw. Trends Mocrobiol. 5:409.

Bowie A, and L. A. O'Neill. 2000. Oxidative stress and nuclear factor-kappaB activation: a reassessment of the evidence in the light of recent discoveries. Biochem. Pharmacol. 59:13.

Brennan, P., and L. A. O'Neill. 1996. 2-mercaptoethanol restores the ability of nuclear factor kappa B (NF kappa B) to bind DNA in nuclear extracts from interleukin 1-treated cells incubated with pyrollidine dithiocarbamate (PDTC). Evidence for oxidation of glutathione in the mechanism of inhibition of NF kappa B by PDTC. Biochem. J. 320:975.

Burke, T. 1968. Dengue haemorrhagic fever: a pathological study. Trans. R. Soc. Trop. Med. Hyg. 62:682.

Cammack, N., and Gould, E. A. 1986. Antigenic analysis of yellow fever virus glycoproteins: use of monoclonal antibodies in enzyme-linked immunosorbent assays. J. Virol. Methods 13: 1699-1709.

Chambers, T. J., D. W. McCourt, and C. M. Rice. 1990. Production of yellow fever virus protein in infected cells: identification of discrete polyprotein species and analysis of cleavage. Virology 177:159.

Chang H. H., H. F. Shyu, Y. M. Wang, D.S. Sun, R. H. Shyu, S. S. Tang, Y. S. Huang. 2002. Facilitation of cell adhesion by immobilized dengue viral nonstructural protein 1 (NS1): arginine-glycine-aspartic acid structural mimicry within the dengue viral NS1 antigen. J. Infect. Dis. 186:743

Chaturvedi, U. C., E. A. Elbishbishi, R. Agarwal, and A. S. Mustafa. 2001. Cytotoxic factor-autoantibodies: posibile role in the pathogenesis of dengue haemorrhagic fever. FEMS Immunol. Med. Microbiol. 30:181.

Chen, Y. C., S. Y. Wang, and C. C. King. 1999. Bacterial lipopolysaccharide inhibits dengue virus infection of primary human monocytes/macrophages by blockade of virus entry via a CD14-dependent mechanism. J. Virol. 73:2650.

Falconar, A. K. I. 1997. The dengue virus nonstructural-1 protein (NS1) generates antibodies to common epitopes on human blood clotting, integrin/adhesin proteins and binds to human endothelial cells: potential implications in haemorrhagic fever pathogenesis. Arch. Virol. 142:897.

Falgout, B., R. Chanock, and C. J. Lai. 1989. Proper processing of dengue virus nonstructural glycoprotein NS1 requires the N-terminal hydrophobic signal sequence and the downstream nonstructural protein NS2A. J. Virol. 63:1852.

Falgout, B., M. Bray, J. J. Schlesinger, and C. J. Lai. 1990. Immunization of mice with recombinant vaccinia virus expressing authentic dengue virus NS1 protect against lethal dengue virus encephalitis. J. Virol. 64:4356.

Falgout, B., M. Pethel, Y. M. Zhang, and C. J. Lai. 1991. Both nonstructural proteins NS2B and NS3 are required for the proteolytic processing of dengue virus nonstructural proteins. J. Virol. 65:2467.

Fonseca, B. A. L., S. Pincus, R. E. Shope, E. Paoletti, and P. W. Mason. 1994. Recombinant vaccinia viruses co-expressing dengue-1 glycoproteins prM and E induce neutralizing antibodies in mice. Vaccine 12:279.

Gagnon, S. J., F. A. Ennis, and A. L. Rothman. 1999. Bystander target cell lysis and cytokine production by dengue virus-specific human CD4+ cytotoxic T-lymphocyte clones. J. Virol. 73:3623.

Gibson, A. W., J. H. Kim, L. A. Wojciechowicz, B. Zhang, C. A. Michels, and C. H. Kim. 1999. Pyrithione, a zinc ionophore, inhibits NF-kappaB activation. Curr. Genetics 36:1.

Gorbalenya, A. E., A. P. Donchenko, E. V. Koonin, and V. M. Blinov. 1989. N-terminal domains of putative helicases of flavi- and pestiviruses may be serine proteases. Nucleic Acids Res. 17:3889.

Grilli, M., J. J. S. Chin, and M. J. Lenardo. 1993. NF-kB and Rel: Participants in a multiform transcriptional regulatory system. Int. Rev. Cytol. 143:1.

Gubler, D. J. 1998. Dengue and dengue haemorrahgic fever. Clin. Mocrobiol. Rev. 11:480.

Guirakhoo F, J. Arroyo, K. V. Pugachev, C. Miller, Z. X. Zhang, R. Weltzin, K. Georgakopoulos, J. Catalan, S. Ocran, K. Soike, M. Ratterree, T. P. Monath. 2001. Construction, safety, and immunogenicity in nonhuman primates of a chimeric yellow fever-dengue virus tetravalent vaccine. J. Viol. 75:7290

Hall, W. C., T. P. Crowell, D. E. Watts, V. L. Barros, H. Kruger, F. Pinheiro, and C. J. Peters. 1991. Demonstration of yellow fever and dengue antigens in formalin-fixed paraffin-embedded human liver by immunohistochemical analysis. Am. J. Trop. Med. Hyg. 45:408.

Halstead, S. B. 1970. Observations related to pathogenesis of dengue hemorrhagic fever. VI. Hypotheses and discussion. Yale J. Biol. Med. 42:350.

Halstead, S. B., and E. J. O’Rourke. 1977. Antibody-enhanced dengue virus infection in primate leukocytes. Nature 265:739.

Halstead, S. B. 1979. In vivo enhancement of dengue virus infection in rhesus monkeys by passively transferred antibody. J. Infect. Dis. 140:527.

Halstead, S. B. 1988. Pathogenesis of dengue: challenges to molecular biology. Science 239:476.

Halstead, S. B. 1989. Antibody, macrophages, dengue virus infection, shock and hemorrhage: a pathogenetic cascade. Rev. Infect. Dis. 11:830.

Halstead, S. B. and Deen J. 2002. The future of dengue vaccines. Lancet. 360:1243.

Henchal, E. A., L. S. Henchal, and J. J. Schlesinger. 1988. Synergistic interactions of anti-NS1 monoclonal antibodies protect passively immunized mice from lethal challenge with dengue-2 virus. J. Gen. Virol. 209:2101.

Henchal, E. A., and J. R. Putnak. 1990. The dengue virus. Clin. Micro. Rev. 3:376.

Huang, K. J., S. Y. J. Li, S. C. Chen, H. S. Liu, Y. S. Lin, T. M. Yeh, C. C. Liu, and H. Y. Lei. 2000. Manifestation of thrombocytopenia in dengue-2-virus-infected mice. J. Gen. Virol. 81:2177.

Kalayanarooj, S., D. W. Vaughn, S. Nimmannitya, S. Green, S. Suntayakorn, N. Kunentrasai, W. Viramitrachai, S. Ratanachueke, S. Kiatpolpoj, B. L. Innis, A. L. Rothman, A. Nisalak, and F. A. Ennis. 1997. Early clinical and laboratory indictors of acute dengue illness. J. Infect. Dis. 176:313.

Kim, S., H. S. Ro, and C. H. Kim. 1999. Zinc is required in pyrrolidine dithiocarbamate inhibition of NF-kappaB activation. J. Biol. Chem. 274:14678.

King, C. A., J. S. Marshall, H. Alshurafa, and R. Anderson. 2000. Release of vasoactive cytokines by antibody-enhanced dengue virus infection of a human mast cell/basophil line. J. Virol. 74:7146.

Kochel, T., S. J. Wu, K. Raviprakash, P. Hobart, S. Hoffman, K. Porter, and C. Hayes. 1997. Inoculation of plasmids expressing the dengue-2 envelope gene elicit neutralizing antibodies in mice. Vaccine 15:547.

Kuo, C. H., D. I. Tai, C. S. Chang-Chien, C. K. Lan, S. S. Chiou, and Y. F. Liaw. 1992. Liver biochemical tests and dengue fever. Am. J. Trop. Med. Hyg. 47:265.

Lalor PF, P. Shields, A. Grant, D. H. Adams. 2002. Recruitment of lymphocytes to the human liver. Immunol. Cell. Biol. 80:52.

Lakshminarayanan, V., E. A. Drab-Weiss, and K. A. Roebuck. 1998. H2O2 and tumor necrosis factor-alpha induce differential binding of the redox-responsive transcription factors AP-1 and NF-kappaB to the interleukin-8 promoter in endothelial and epithelial cells. J. Biol. Chem. 273:32670.

Lakshminarayanan, V., D. W. Beno, R. H. Costa, and K. A. Roebuck. 1997. Differential regulation of interleukin-8 and intercellular adhesion molecule-1 by H2O2 and tumor necrosis factor-alpha in endothelial and epithelial cells. J. Biol. Chem. 272:32910.

Lei, H. Y., T. M. Yeh, H. S. Liu, Y. S. Lin, S. H. Chen, and C. C. Liu. 2001. Immunopathogenesis of dengue virus infection. J. Biomed. Sci. 8:377.

Lin, C. F., H. Y. Lei, C. C. Liu, H. S. Liu, T. M. Yeh, S. T. Wang, T. I. Yang, F. C. Sheu, C. F. Kuo, and Y. S. Lin. 2001. Generation of IgM anti-platelet autoantibody in dengue patients. J. Med. Virol. 63:143-149.

Lin, C. F., H. Y. Lei, A. L. Shiau, H. S. Liu, T. M. Yeh, S. H. Chen, C. C. Liu, S. C. Chiu, and Y. S. Lin. 2002. Endothelial cell apoptosis induced by antibodies against dengue virus nonstructural protein 1 via production of nitric oxide. J. Immunol. 169:657

Lin, C. F., H. Y. Lei, A. L. Shiau, C. C. Liu, H. S. Liu, T. M. Yeh, S. H. Chen, and Y. S. Lin. 2003. Antibodies from dengue patient sera cross-react with endothelial cells and induce damage. J. Med. Virol. 69:82

Lin, Y. L., L. K. Chen, C. L. Liao, C. T. Yeh, S. H. Ma, J. L. Chen, Y. L. Huang, S. S. Chen, and H. Y. Chiang. 1998. DNA immunization with Japanese encephalitis virus nonstructural protein NS1 elicit protective immunity in mice. J. Virol. 72:191.

Lin, Y. L., C. C. Liu, H. Y. Lei, T. M. Yeh, Y. S. Lin, R. M. Y. Chen, and H. S. Liu. 2000a. Infection of five human liver cell lines by dengue-2 virus. J. Med. Virol. 60:425.

Lin, Y. L., C. C. Liu, J. I. Chuang, H. Y. Lei, T. M. Yeh, Y. S. Lin, Y. H. Huang, and H. S. Liu. 2000b. Involvement of oxidative stress, NF-IL-6, and RANTES expression in dengue-2-virus-infected human liver cells. Virology 276:114.

Liu C. C., K. J. Huang, Y. S. Lin, T. M. Yeh, H. S. Liu, and H. Y. Lei. 2002. Transient CD4/CD8 ratio inversion and aberrant immune activation during dengue virus infection. J. Med. Virol. 68:241.

Marianneau, P., A. Cardona, L. Edelman, V. Deubel, and P. Despres. 1997. Dengue virus replication in human hepatoma cells activates NF-kB which in turn induces apoptotic cell death. J. Virol. 71:3244.

Markoff L, X. Pang, H. S. Houng Hs, B. Falgout, R. Olsen, E. Jones, S. Polo. 2002. Derivation and characterization of a dengue type 1 host range-restricted mutant virus that is attenuated and highly immunogenic in monkeys. J. Virol. 76:3318.

Mathew, A., I. Kurane, S. Green, D. W. Vaughn, S. Kalayanarooj, S. Suntayakorn, F. A. Ennis, and A. L. Rothman. 1999. Impaired T cell proliferation in acute dengue infection. J. Immunol. 162:5609.

Men, R. H., M. Bray, and C. J. Lai. 1991. Carboxy-terminally truncated dengue virus envelope glycoproteins expressed on the cell surface and secreted extracellularly exhibit increased immunogenicity in mice. J. Virol. 65:1400.

Mongkolsapaya J, W. Dejnirattisai, X. N. Xu, S. Vasanawathana, N. Tangthawornchaikul, A. Chairunsri, S. Sawasdivorn, T. Duangchinda, T. Dong, S. Rowland-Jones, P. T. Yenchitsomanus, A. McMichael, P. Malasit, G. Screaton. 2003 Original antigenic sin and apoptosis in the pathogenesis of dengue hemorrhagic fever. Nat. Med. 9:921.

Mukerjee, R., U. C. Chaturvedi, and R. Dhawan. 1995. Dengue virus-induced human cytotoxic factor: production by peripheral blood leukocytes in vitro. Clin. Exp. Immunol. 102:262.

Munoz, C., M. C. Castellanos, A. Alfranca, A. Vara, M. A. Esteban, J. M. Redondo, M. O. de Landazuri. 1996. Transcriptional up-regulation of intracellular adhesion molecule-1 in human endothelial cells by the antioxidant pyrrolidine dithiocarbamate involves the activation of activating protein-1. J. Immunol. 157:3587.

O’Neill L. A., and C. Kaltschmidt. 1997. NFkB: A crucial transcription factor for glial and neuronal cell function. Trends Neurosci. 20:252.

Preugschat, F., C. W. Yao, and J. H. Strauss. 1990. In vitro processing of dengue virus type 2 nonstructural proteins NS2A, NS2B, and NS3. J. Virol. 64:4364.

Preugschat, F., and J. H. Strauss. 1991. Processing of nonstructural proteins NS4A and NS4B of dengue 2 virus in vitro and in vivo. Virology 185:689.

Rhandolph, V. B., G. Winkler, and V. Stollar. 1990. Acidotropic amines inhibit proteolytic processing of flavivirus prM protein. Virology 174:450.

Rice, A. P., M. Kostura, and M. B. Mathews. 1989. Identification of a 90-kDa polypeptide which associates with adenovirus VA RNAI and is phosphorylated by the double-strand RNA-dependent protein kinase. J. Biol. Chem. 264:20632.

Rosen, L. 1977. The emperor’s new clothes revisited, or reflections on the pathogenesis of dengue hemorrhagic fever. Am. J. Trop. Med. Hyg. 26:337.

Schlesinger, J. J., M. W. Brandriss, and E. E. Walsh. 1987. Protection of mice against dengue 2 virus encephalitis by immunization with the dengue 2 virus non-structural glycoprotein NS1. J. Gen. Virol. 68:853.

Schmidt, K. N., P. Amstad, P. Cerutti, and P. A. Baeuerle. 1996. Identification of hydrogen peroxide as the relevant messenger in the activation pathway of transcription factor NF-kappaB. Adv. Exp. Med. Biol. 387:63.

Schutze, S., K. Potthoff, T. Machleidt, D. Berkovic, K. Weigmann, M. Kronke. 1992. TNF activates NF-kB by phosphatidylcholine-specific phospholipase C-induced “acidic” sphingomyelin breakdown. Cell 71:765.

Srivastava, A. K., J. R. Putnak, R. L. Warren, and C. H. Jr. Hoke. 1995. Mice immunized with a dengue type 2 virus E and NS1 fusion protein made in Escherichia coli are protected against lethal dengue virus infection. Vaccine 13:1251.

Staropoli, I., M. P. Frenkiel, F. Megret, and V. Deubel. 1997. Affinity-purified dengue-2 virus envelope glycoprotein induces neutralizing antibodies and protective immunity in mice. Vaccine 15:1946.

Suzuki, Y. J., B. B. Aggarwal, and L. Packer. 1992. Alpha-lipoic acid is a potent inhibitor of NF-kappa B activation in human T cells. Biochem., Biophys. Res. Commun. 189:1709.

Taekema-Roelvink ME, C. Kooten, S. V. Kooij, E. Heemskerk, M. R. Daha 2002. Proteinase 3 enhances endothelial monocyte chemoattractant protein-1 production and induces increased adhesion of neutrophils to endothelial cells by upregulating intercellular cell adhesion molecule-1. J. Am. Soc. Nephrol. 12:932.

Vaughn, D. W., S. Green, S. Kalayanarooj, B. L. Innis, S. Nimmannitya, S. Suntayakorn, T. P. Endy, B. Raengsakulrach, A. L. Rothman, F. A. Ennis, and A. Nisalak. 2000. Dengue viremia titer, antibody response pattern, and virus serotype correlate with disease severity. J. Infect. Dis. 181:2.

Velzing, J., J. Groen, M. T. Drouet, G. van Amerongen, C. Copra, A. D. M. E. Osterhaus, and V. Deubel. 1999. Induction of protective immunity against dengue virus type 2: comparison of candidate live attenuated and recombinant vaccines. Vaccine 17:1312.

Venugopal, K., and E. A. Gould. 1994. Towards a new generation of flavivirus vaccines. Vaccine 12:966.

Viedt, C, R. Dechend, J. Fei, G. M. Hansch, J. Kreuzer, S. R. Orth. 2002. MCP-1 induces inflammatory activation of human tubular epithelial cells: involvement of the transcription factors, nuclear factor-kappaB and activating protein-1. J. Am. Soc. Nephrol. 13:1534.

Wang, L. Y., W. Y. Chang, S. N. Lu, and T. P. Chen. 1990. Sequential changes of serum transaminase and abnormal sonography in patients with suspected dengue fever. Kao Hsiung I Hsueh Ko Hsueh Tsa Chih 6:483.

Wang, S., R. He, J. Patarapotikul, B. L. Innis, and R. Anderson. 1995. Antibody-enhanced binding of dengue-2 virus to human platelets. Virology 213:254.

Winkler, G., V. B. Randolph, G. R. Cleaves, T. E. Ryan, Stollar V. 1988. Evidence that the mature form of Flavivirus nonstructural protein NS1 in dimer. Virology 162:187.

Winkler, G., Maxwell S. E, Ruemmler C., Stollar V. 1989. Newly synthesized dengue-2 virus nonstructural protein NS1 is a soluble protein but becomes partially hydrophobic and membrane-associated after dimerization. Virology 171:302.

Zhang Y, J. Corver, P. R. Chipman, W. Zhang, S. V. Pletnev, D. Sedlak, T.S. Baker, J.H. Strauss, R.J. Kuhn, M.G. Rossmann. 2003. Structures of immature flavivirus particles. EMBO. J. 22:2604.
論文全文使用權限
  • 同意授權校內瀏覽/列印電子全文服務,於2003-08-28起公開。
  • 同意授權校外瀏覽/列印電子全文服務,於2003-08-28起公開。


  • 如您有疑問,請聯絡圖書館
    聯絡電話:(06)2757575#65773
    聯絡E-mail:etds@email.ncku.edu.tw