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研究生:林仕偉
研究生(外文):Shi-WeiLin
論文名稱:登革病毒非結構蛋白1抑制凝血酶原活化以及製造其單株抗體來偵測登革病毒感染
論文名稱(外文):Dengue virus nonstructural protein 1 inhibits prothrombin activation and generation of its monoclonal antibodies as a tool to detect dengue virus infection
指導教授:葉才明
指導教授(外文):Trai-Ming Yeh
學位類別:碩士
校院名稱:國立成功大學
系所名稱:醫學檢驗生物技術學系碩博士班
學門:醫藥衛生學門
學類:醫學技術及檢驗學類
論文種類:學術論文
論文出版年:2012
畢業學年度:100
語文別:中文
論文頁數:118
中文關鍵詞:凝血登革病毒非結構蛋白1凝血酶原凝血酶單株抗體
外文關鍵詞:coagulationDENVNS1thrombinprothrombinmonoclonal antibody
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登革病毒 (DENV) 是一種經由節肢動物傳播的致病原,透過埃及斑蚊和白線斑蚊的叮咬而感染人類,主要發生在熱帶和亞熱帶國家。登革病毒感染造成的疾病,從較輕微的登革熱 (DF),到嚴重的登革出血熱 (DHF) 和登革休克症候群 (DSS)。登革病毒非結構蛋白1 (NS1) 是一個分子量43 kDa的醣蛋白,以二聚體 (dimer) 的型式表現在感染的細胞表面上,或是以六聚體 (hexamer) 的型式分泌到登革病患的血清中。目前為止,雖然對於NS1的確切功能還不是很清楚,但是最近有研究指出NS1能夠結合上補體系統當中的補體4 (C4) 和C4結合蛋白 (C4b binding protein),並且抑制補體的活化。除此之外,分泌出來的NS1可以作為早期診斷登革感染的生物標記。在本研究中,我們利用酵素連結免疫分析法發現在登革病人血清中的NS1可以結合凝血因子II (凝血酶) 以及有NS1和凝血酶的複合物存在,並進一步使用重組NS1 (rNS1) 藉由酵素連結免疫分析法、蛋白質交互作用沉澱試驗 (pull down assay) 和rNS1親和性管柱層析法來證實NS1與凝血酶原或凝血酶之間的結合。雖然rNS1可以結合凝血酶卻不影響凝血酶的活性,然而rNS1會去抑制凝血酶原的活化,使活性部分凝血酶時間 (APTT) 延長,並用抑制凝血酶原活化的呈色試驗來證明。綜合來說,這些實驗結果表示NS1與凝血酶原的結合,並抑制其凝血酶原活化,而導致在DHF/DSS的病患中有出血的情形。依據NS1可以結合凝血酶原和凝血酶,我們更進一步製造抗NS1之單株抗體來偵測登革病人血清內NS1與凝血酶的複合物。我們將rNS1免疫BALB/c小鼠利用融合瘤技術產生專一性高的NS1單株抗體,最後得到七株單株抗體藉由酵素連結免疫分析法和流式細胞儀都可以辨認到rNS1以及登革病毒感染的細胞,這些單株抗體未來可以應用到偵測登革病人體內的NS1。
Dengue virus (DENV) is an arthropod-borne human pathogen, which infects human through mosquito Aedes albopictus and Aedes aegypti and mainly occurs in tropical and subtropical countries. DENV infection can cause disease from mild febrile to severe dengue hemorrhage fever (DHF) and dengue shock syndrome (DSS). Dengue nonstructural protein 1 (NS1) is a 43 kDa glycoprotein which is expressed as a dimer on infected cell surface or secreted in hexamer form in DENV-infected patient’s serum. Secreted NS1 can be a diagnostic marker for dengue infection. However, the precise function of NS1 is littlely known, except recently it is found that NS1 can bind to complement proteins C4 and C4b binding protein (C4BP) and inhibit complement pathway activation. In this study we found NS1 could bind to coagulation factor II (thrombin) and form NS1-thrombin complex in dengue patients’ sera by enzyme-linked immunosorbent assay (ELISA). The binding of NS1 to prothrombin or thrombin was further confirmed, using recombinant NS1 (rNS1) and different assays such as ELISA, pull-down assay and rNS1-affinity column purification. Furthermore, binding of rNS1 to prothrombin inhibited prothrombin activation which was demonstrated by prolongation of activated partial thromboplastin time and inhibition of prothrombin activation-induced chromogenic change. Taken together, these results suggest NS1 can bind to prothrombin and inhibit its activation, which may contribute to the hemorrhage in DHF/DSS patients. Based on the finding that NS1 can bind to prothrombin/thrombin, we further generated anti-NS1 monoclonal antibodies (mAbs) to detect NS1-thrombin complex in dengue patients’ sera. We used rNS1 to immunize BALB/c mice to generate mAbs specifically against NS1 by hybridoma technology. We got 7 mAbs which could recognize rNS1 and DENV-infected cells by ELISA and flow cytometry. These mAbs can be applied in the future to detect NS1 in DENV infected samples.
中文摘要..I
英文摘要..III
誌謝..V
總目錄..VI
表目錄..XI
圖目錄.. XII
縮寫指引..XIII
1. 緒論..1
1.1 登革病毒..1
1.1.1 登革病毒概論與現況..1
1.1.2 登革病毒結構和非結構蛋白..2
1.1.3 登革病毒感染引起的病症及診斷方法..6
1.1.4 登革病毒致病機轉..8
1.2 血液恆定機制..11
1.3 單株抗體的發展及應用..15
2. 實驗動機與目標..18
3. 實驗材料與方法..19
3.1 登革病人和控制組血清來源..19
3.2 登革病毒之製備..19
3.2.1 登革病毒來源..19
3.2.2 細胞株..19
3.2.2.1 C6/36 cell line..19
3.2.2.2 Baby hamster kidney-21 (BHK-21) cell line..20
3.2.3 培養登革病毒...20
3.2.4 定量登革病毒效價..20
3.3 構築登革病毒 Nonstructural protein 1 (NS1) 重組蛋白..21
3.3.1 萃取登革病毒RNA..21
3.3.2 反轉錄聚合酵素連鎖反應 (RT-PCR)..22
3.3.3 聚合酵素連鎖反應 (PCR)..23
3.3.4 製備勝任細胞 (competent cell)..25
3.3.5 質體構築 (construction) 及轉型 (transformation) ..26
3.4 表現登革病毒Nonstructural protein 1 (NS1) 重組蛋白.. 29
3.4.1 重組蛋白的表現及純化..29
3.4.2 SDS-PAGE分析重組蛋白..31
3.4.3 西方墨點法分析重組蛋白..32
3.5 登革病毒 NS1重組蛋白的功能分析..33
3.5.1 NS1重組蛋白結合血液凝固因子凝血酶原和凝血酶..33
3.5.1.1 酵素連結免疫分析法 (ELISA)..33
3.5.1.2 NS1和凝血酶交互作用沉澱試驗 (pull down assay) ..35
3.5.1.3 NS1重組蛋白親和性層析管柱..35
3.5.2 NS1重組蛋白對血液凝固時間的影響..37
3.5.2.1 測定凝血酶時間 (Thrombin time) ..37
3.5.2.2 測定活化部分凝血酶時間 (Activated partial thromboplastin time)..37
3.5.2.3 測定凝血酶原時間 (Prothrombin time).. 38
3.5.3 NS1重組蛋白對凝血酶原和凝血酶的活性測試..38
3.6 登革病毒NS1抗體的製作..39
3.6.1 動物來源..39
3.6.2 免疫BALB/c小鼠及血清製備..39
3.6.3 血清抗體效價的測定..39
3.7 登革病毒NS1單株抗體的製備..40
3.7.1 準備骨髓瘤細胞 (Myeloma cell)..41
3.7.2 前處理及融合..41
3.7.3 篩選..42
3.7.4 獲取 (Harvest)..42
3.7.5 大量製備抗體及純化..43
3.8 登革病毒NS1單株抗體分析..44
3.8.1 NS1單株抗體之抗體分型測定..44
3.8.2 利用西方墨點法測試登革病毒NS1單株抗體之結合能力..44
3.8.3 測試NS1單株抗體結合至native form NS1之能力..45
3.8.3.1 利用流式細胞儀測試單株抗體結合能力..45
3.8.3.2 利用免疫螢光染色測試單株抗體結合能力..46
3.9 發展capture ELISA偵測登革病毒NS1蛋白..47
3.9.1 利用登革病毒NS1單株抗體偵測重組蛋白NS1能力.. 47
3.9.2 利用capture ELISA偵測感染登革病毒之細胞裂解物與上清液..48
3.9.3 利用commerical Bio-rad platelia證明感染登革病毒之細胞有表現出native form NS1..48
3.10 統計分析..48
4. 結果..50
4.1 登革病毒非結構蛋白1 (NS1) 造成凝血功能缺失..50
4.1.1 登革病人血清內NS1與凝血酶的結合..50
4.1.2 構築與表現及純化登革病毒NS1重組蛋白..51
4.1.3 登革病毒NS1重組蛋白結合凝血酶原和凝血酶..52
4.1.4 登革病毒NS1重組蛋白對凝血酶的影響..53
4.1.5 登革病毒NS1重組蛋白對凝血酶原的影響..53
4.2 發展NS1單株抗體作為快速檢測登革病毒感染的工具..54
4.2.1 免疫小鼠及製造登革病毒NS1單株抗體..54
4.2.2 NS1單株抗體特性..55
4.2.3 發展NS1抗原捕捉酵素連結免疫分析法..56
4.2.4 以NS1 capture ELISA偵測感染登革病毒細胞表現出的native form NS1..57
5. 討論..58
5.1 重組非結構蛋白1 (NS1) 的製備..58
5.2 NS1結合凝血酶原與凝血酶..59
5.3 NS1對凝血酶和凝血酶原的影響..60
5.4 登革病毒NS1單株抗體..63
6. 結論..67
7. 引用文獻..68
8. 表附錄..83
9. 圖附錄..86
附錄一、血液凝固系統示意圖..109
附錄二、凝血酶原活化成凝血酶過程示意圖..110
附錄三、實驗耗材、試劑及抗體..111
附錄四、儀器..116
作者簡歷..118
表目錄
Table 1. Statistical analysis of NS1 binding to thrombin and NS1-thrombin complex in dengue patients’ sera.
Table 2. Comparison of NS1-thrombin complex and commercial NS1 detection in dengue patients’ sera.
Table 3. Summary of the properties of anti-NS1 monoclonal antibodies.
圖目錄
Figure 1. Detection of NS1 binding to thrombin and NS1-thrombin complex in patients' sera.
Figure 2. Construction of DENV 2 PL046 NS1 expression vector.
Figure 3. Purification and characterization of rNS1.
Figure 4. Binding of rNS1 to thrombin and prothrombin.
Figure 5. Effect of rNS1 on thrombin activity as determined by thrombin time and chromogenic assay.
Figure 6. rNS1 prolongs APTT and inhibits prothrombin conversion to thrombin.
Figure 7. rNS1 didn’t prolong prothrombin time.
Figure 8. Analysis of rNS1-immunized mice sera.
Figure 9. Selection of anti-NS1 monoclonal antibodies.
Figure 10. The binding ability of mAbs to DENV-infected cells analyzed by flow cytometry and IFA.
Figure 11. Development of NS1 capture ELISA.
Figuee 12. Detection of native form NS1 by NS1 capture ELISA.
Figure 13. Confirmation of native form NS1 existed in DENV 2 infected C6/36 lysate (A) and supernatant (B) using commercial Bio-rad platelia dengue NS1 antigen kit.

Alcon-LePoder, S., P. Sivard, et al. (2006). Secretion of flaviviral non-structural protein NS1: from diagnosis to pathogenesis. Novartis Found Symp 277: 233-247; discussion 247-253.
Alcon, S., A. Talarmin, et al. (2002). Enzyme-linked immunosorbent assay specific to Dengue virus type 1 nonstructural protein NS1 reveals circulation of the antigen in the blood during the acute phase of disease in patients experiencing primary or secondary infections. J Clin Microbiol 40(2): 376-381.
Allison, S. L., J. Schalich, et al. (2001). Mutational evidence for an internal fusion peptide in flavivirus envelope protein E. J Virol 75(9): 4268-4275.
Allonso, D., M. da Silva Rosa, et al. (2011). Polyclonal antibodies against properly folded Dengue virus NS1 protein expressed in E. coli enable sensitive and early dengue diagnosis. J Virol Methods 175(1): 109-116.
Amberg, S. M., A. Nestorowicz, et al. (1994). NS2B-3 proteinase-mediated processing in the yellow fever virus structural region: in vitro and in vivo studies. J Virol 68(6): 3794-3802.
Arya, S. C. and N. Agarwal (2007). Evaluation of immunoglobulin M and G capture enzyme-linked immunosorbent assay Panbio kits for diagnostic dengue infections. Journal of Clinical Virology 40(1): 89-89.
Avirutnan, P., A. Fuchs, et al. (2010). Antagonism of the complement component C4 by flavivirus nonstructural protein NS1. Journal of Experimental Medicine 207(4): 793-806.
Avirutnan, P., R. E. Hauhart, et al. (2011). Binding of Flavivirus Nonstructural Protein NS1 to C4b Binding Protein Modulates Complement Activation. J Immunol 187(1): 424-433.
Avirutnan, P., N. Punyadee, et al. (2006). Vascular leakage in severe dengue virus infections: a potential role for the nonstructural viral protein NS1 and complement. J Infect Dis 193(8): 1078-1088.
Avirutnan, P., L. Zhang, et al. (2007). Secreted NS1 of dengue virus attaches to the surface of cells via interactions with heparan sulfate and chondroitin sulfate E. PLoS Pathog 3(11): e183.
Azeredo, E. L., S. M. Zagne, et al. (2001). Characterisation of lymphocyte response and cytokine patterns in patients with dengue fever. Immunobiology 204(4): 494-507.
Bachmann, F. and I. E. Kruithof (1984). Tissue plasminogen activator: chemical and physiological aspects. Semin Thromb Hemost 10(1): 6-17.
Bessoff, K., E. Phoutrides, et al. (2010). Utility of a commercial nonstructural protein 1 antigen capture kit as a dengue virus diagnostic tool. Clin Vaccine Immunol 17(6): 949-953.
Bradbury, A. R. and J. D. Marks (2004). Antibodies from phage antibody libraries. J Immunol Methods 290(1-2): 29-49.
Bridget, W., V. N. Tran, et al. (2009). Hemostatic Changes in Vietnamese Children with Mild Dengue Correlate with the Severity of Vascular Leakage Rather than Bleeding. American Journal of Tropical Medicine and Hygiene 81(4): 638-644.
Chaiyaratana, W., A. Chuansumrit, et al. (2009). Evaluation of dengue nonstructural protein 1 antigen strip for the rapid diagnosis of patients with dengue infection. Diagn Microbiol Infect Dis 64(1): 83-84.
Chakravarti, A. and R. Kumaria (2006). Circulating levels of tumour necrosis factor-alpha & interferon-gamma in patients with dengue & dengue haemorrhagic fever during an outbreak. Indian J Med Res 123(1): 25-30.
Chen, L. C., H. Y. Lei, et al. (2006). Correlation of serum levels of macrophage migration inhibitory factor with disease severity and clinical outcome in dengue patients. Am J Trop Med Hyg 74(1): 142-147.
Chen, S. T., Y. L. Lin, et al. (2008). CLEC5A is critical for dengue-virus-induced lethal disease. Nature 453(7195): 672-676.
Chen, Y., T. Maguire, et al. (1997). Dengue virus infectivity depends on envelope protein binding to target cell heparan sulfate. Nat Med 3(8): 866-871.
Chuang, Y. C., H. Y. Lei, et al. (2011). Dengue virus-induced autoantibodies bind to plasminogen and enhance its activation. J Immunol 187(12): 6483-6490.
Chuansumrit, A., C. Puripokai, et al. (2010). Laboratory predictors of dengue shock syndrome during the febrile stage. Southeast Asian J Trop Med Public Health 41(2): 326-332.
Chungue, E., O. Cassar, et al. (1995). Molecular epidemiology of dengue-1 and dengue-4 viruses. J Gen Virol 76 ( Pt 7): 1877-1884.
Chungue, E., L. Poli, et al. (1994). Correlation between detection of plasminogen cross-reactive antibodies and hemorrhage in dengue virus infection. J Infect Dis 170(5): 1304-1307.
Clarke, D. H. and J. Casals (1958). Techniques for hemagglutination and hemagglutination-inhibition with arthropod-borne viruses. Am J Trop Med Hyg 7(5): 561-573.
Colman, R. W. (1984). Surface-mediated defense reactions. The plasma contact activation system. J Clin Invest 73(5): 1249-1253.
Cui, T., R. J. Sugrue, et al. (1998). Recombinant dengue virus type 1 NS3 protein exhibits specific viral RNA binding and NTPase activity regulated by the NS5 protein. Virology 246(2): 409-417.
Das, D., S. L. Gares, et al. (2004). Evaluation of a Western Equine Encephalitis recombinant E1 protein for protective immunity and diagnostics. Antiviral Res 64(2): 85-92.
Das, D., F. Jacobs, et al. (2007). Differential expression of the Ebola virus GP(1,2) protein and its fragments in E. coli. Protein Expr Purif 54(1): 117-125.
Diercks, F. H. (1959). Isolation of a type 2 dengue virus by use of hamster kidney cell cultures. Am J Trop Med Hyg 8(4): 488-491.
Dussart, P., B. Labeau, et al. (2006). Evaluation of an enzyme immunoassay for detection of dengue virus NS1 antigen in human serum. Clin Vaccine Immunol 13(11): 1185-1189.
Dussart, P., L. Petit, et al. (2008). Evaluation of two new commercial tests for the diagnosis of acute dengue virus infection using NS1 antigen detection in human serum. PLoS Negl Trop Dis 2(8): e280.
Duyen, H. T., T. V. Ngoc, et al. (2011). Kinetics of plasma viremia and soluble nonstructural protein 1 concentrations in dengue: differential effects according to serotype and immune status. J Infect Dis 203(9): 1292-1300.
Eldadah, Z. A., D. M. Asher, et al. (1991). Detection of flaviviruses by reverse-transcriptase polymerase chain reaction. J Med Virol 33(4): 260-267.
Falgout, B., M. Pethel, et al. (1991). Both nonstructural proteins NS2B and NS3 are required for the proteolytic processing of dengue virus nonstructural proteins. J Virol 65(5): 2467-2475.
Flamand, M., F. Megret, et al. (1999). Dengue virus type 1 nonstructural glycoprotein NS1 is secreted from mammalian cells as a soluble hexamer in a glycosylation-dependent fashion. J Virol 73(7): 6104-6110.
Forwood, J. K., A. Brooks, et al. (1999). The 37-amino-acid interdomain of dengue virus NS5 protein contains a functional NLS and inhibitory CK2 site. Biochem Biophys Res Commun 257(3): 731-737.
Fry, S. R., M. Meyer, et al. (2011). The diagnostic sensitivity of dengue rapid test assays is significantly enhanced by using a combined antigen and antibody testing approach. PLoS Negl Trop Dis 5(6): e1199.
Germi, R., J. M. Crance, et al. (2002). Heparan sulfate-mediated binding of infectious dengue virus type 2 and yellow fever virus. Virology 292(1): 162-168.
Gorbalenya, A. E., E. V. Koonin, et al. (1989). Two related superfamilies of putative helicases involved in replication, recombination, repair and expression of DNA and RNA genomes. Nucleic Acids Res 17(12): 4713-4730.
Guzman, M. G., T. Jaenisch, et al. (2010). Multi-country evaluation of the sensitivity and specificity of two commercially-available NS1 ELISA assays for dengue diagnosis. PLoS Negl Trop Dis 4(8).
Guzman Tirado, M. G., G. Kouri Flores, et al. (1984). [Isolation of dengue virus type 2 from the serum of patients using the suckling mouse and cultured LLCMK2 cells]. Rev Cubana Med Trop 36(1): 4-10.
Halstead, S. B. and E. J. O'Rourke (1977). Dengue viruses and mononuclear phagocytes. I. Infection enhancement by non-neutralizing antibody. J Exp Med 146(1): 201-217.
Halstead, S. B., C. N. Venkateshan, et al. (1984). Heterogeneity of infection enhancement of dengue 2 strains by monoclonal antibodies. J Immunol 132(3): 1529-1532.
Hang, V. T., N. M. Nguyet, et al. (2009). Diagnostic accuracy of NS1 ELISA and lateral flow rapid tests for dengue sensitivity, specificity and relationship to viraemia and antibody responses. PLoS Negl Trop Dis 3(1): e360.
Harris, E., T. G. Roberts, et al. (1998). Typing of dengue viruses in clinical specimens and mosquitoes by single-tube multiplex reverse transcriptase PCR. J Clin Microbiol 36(9): 2634-2639.
Henchal, E. A., L. S. Henchal, et al. (1988). Synergistic interactions of anti-NS1 monoclonal antibodies protect passively immunized mice from lethal challenge with dengue 2 virus. J Gen Virol 69 ( Pt 8): 2101-2107.
Henchal, E. A. and J. R. Putnak (1990). The dengue viruses. Clin Microbiol Rev 3(4): 376-396.
Herrera, M., S. Vazquez, et al. (1985). [Determination of complement fixation antibodies in patients with dengue hemorrhagic fever]. Rev Cubana Med Trop 37(2): 195-202.
Huang, J. L., J. H. Huang, et al. (2001). High-level expression of recombinant dengue viral NS-1 protein and its potential use as a diagnostic antigen. J Med Virol 65(3): 553-560.
Huang, K. J., S. Y. Li, et al. (2000). Manifestation of thrombocytopenia in dengue-2-virus-infected mice. J Gen Virol 81(Pt 9): 2177-2182.
Huang, Y. H., H. Y. Lei, et al. (2003). Tissue plasminogen activator induced by dengue virus infection of human endothelial cells. J Med Virol 70(4): 610-616.
Huang, Y. H., C. C. Liu, et al. (2001). Activation of coagulation and fibrinolysis during dengue virus infection. J Med Virol 63(3): 247-251.
Jacobs, M. G., P. J. Robinson, et al. (2000). Dengue virus nonstructural protein 1 is expressed in a glycosyl-phosphatidylinositol-linked form that is capable of signal transduction. FASEB J 14(11): 1603-1610.
Johnson, B. W., B. J. Russell, et al. (2005). Serotype-specific detection of dengue viruses in a fourplex real-time reverse transcriptase PCR assay. J Clin Microbiol 43(10): 4977-4983.
Kettleborough, C. A., K. H. Ansell, et al. (1994). Isolation of tumor cell-specific single-chain Fv from immunized mice using phage-antibody libraries and the re-construction of whole antibodies from these antibody fragments. Eur J Immunol 24(4): 952-958.
Khor, B. S., J. W. Liu, et al. (2006). Dengue hemorrhagic fever patients with acute abdomen: clinical experience of 14 cases. Am J Trop Med Hyg 74(5): 901-904.
Khromykh, A. A., A. N. Varnavski, et al. (2001). Coupling between replication and packaging of flavivirus RNA: evidence derived from the use of DNA-based full-length cDNA clones of Kunjin virus. J Virol 75(10): 4633-4640.
Kiermayr, S., R. M. Kofler, et al. (2004). Isolation of capsid protein dimers from the tick-borne encephalitis flavivirus and in vitro assembly of capsid-like particles. J Virol 78(15): 8078-8084.
Kohler, G. and C. Milstein (1975). Continuous cultures of fused cells secreting antibody of predefined specificity. Nature 256(5517): 495-497.
Kumarasamy, V., A. H. Wahab, et al. (2007). Evaluation of a commercial dengue NS1 antigen-capture ELISA for laboratory diagnosis of acute dengue virus infection. J Virol Methods 140(1-2): 75-79.
Kuno, G., D. J. Gubler, et al. (1985). Comparative sensitivity of three mosquito cell lines for isolation of dengue viruses. Bull World Health Organ 63(2): 279-286.
Kurucz, I., J. A. Titus, et al. (1995). Correct disulfide pairing and efficient refolding of detergent-solubilized single-chain Fv proteins from bacterial inclusion bodies. Mol Immunol 32(17-18): 1443-1452.
Lanciotti, R. S., C. H. Calisher, et al. (1992). Rapid detection and typing of dengue viruses from clinical samples by using reverse transcriptase-polymerase chain reaction. J Clin Microbiol 30(3): 545-551.
Lanciotti, R. S., D. J. Gubler, et al. (1997). Molecular evolution and phylogeny of dengue-4 viruses. J Gen Virol 78 ( Pt 9): 2279-2284.
Lanciotti, R. S., J. G. Lewis, et al. (1994). Molecular evolution and epidemiology of dengue-3 viruses. J Gen Virol 75 ( Pt 1): 65-75.
Lee, I. K., J. W. Liu, et al. (2005). Clinical characteristics and risk factors for concurrent bacteremia in adults with dengue hemorrhagic fever. Am J Trop Med Hyg 72(2): 221-226.
Lei, H. Y., T. M. Yeh, et al. (2001). Immunopathogenesis of dengue virus infection. J Biomed Sci 8(5): 377-388.
Libraty, D. H., P. R. Young, et al. (2002). High circulating levels of the dengue virus nonstructural protein NS1 early in dengue illness correlate with the development of dengue hemorrhagic fever. J Infect Dis 186(8): 1165-1168.
Lima Mda, R., R. M. Nogueira, et al. (2010). Comparison of three commercially available dengue NS1 antigen capture assays for acute diagnosis of dengue in Brazil. PLoS Negl Trop Dis 4(7): e738.
Lin, C. F., H. Y. Lei, et al. (2001). Generation of IgM anti-platelet autoantibody in dengue patients. J Med Virol 63(2): 143-149.
Lin, C. F., H. Y. Lei, et al. (2003). Antibodies from dengue patient sera cross-react with endothelial cells and induce damage. J Med Virol 69(1): 82-90.
Lin, C. F., H. Y. Lei, et al. (2002). Endothelial cell apoptosis induced by antibodies against dengue virus nonstructural protein 1 via production of nitric oxide. J Immunol 169(2): 657-664.
Lin, Y. J. and S. C. Wu (2005). Histidine at residue 99 and the transmembrane region of the precursor membrane prM protein are important for the prM-E heterodimeric complex formation of Japanese encephalitis virus. J Virol 79(13): 8535-8544.
Lin, Y. L., H. Y. Lei, et al. (2002). Heparin inhibits dengue-2 virus infection of five human liver cell lines. Antiviral Res 56(1): 93-96.
Liu, I. J., C. Y. Chiu, et al. (2011). Molecular mimicry of human endothelial cell antigen by autoantibodies to nonstructural protein 1 of dengue virus. J Biol Chem 286(11): 9726-9736.
Liu, L. W., J. Ye, et al. (1991). Proteolytic formation of either of the two prothrombin activation intermediates results in formation of a hirugen-binding site. J Biol Chem 266(35): 23633-23636.
Loskutoff, D. J., J. A. van Mourik, et al. (1983). Detection of an unusually stable fibrinolytic inhibitor produced by bovine endothelial cells. Proc Natl Acad Sci U S A 80(10): 2956-2960.
Mackenzie, J. M., M. K. Jones, et al. (1996). Immunolocalization of the dengue virus nonstructural glycoprotein NS1 suggests a role in viral RNA replication. Virology 220(1): 232-240.
Martina, B. E., P. Koraka, et al. (2009). Dengue virus pathogenesis: an integrated view. Clin Microbiol Rev 22(4): 564-581.
Munoz-Jordan, J. L., M. Laurent-Rolle, et al. (2005). Inhibition of alpha/beta interferon signaling by the NS4B protein of flaviviruses. J Virol 79(13): 8004-8013.
Munoz-Jordan, J. L., G. G. Sanchez-Burgos, et al. (2003). Inhibition of interferon signaling by dengue virus. Proc Natl Acad Sci U S A 100(24): 14333-14338.
Nemerson, Y. and B. Furie (1980). Zymogens and cofactors of blood coagulation. CRC Crit Rev Biochem 9(1): 45-85.
Nybakken, G. E., T. Oliphant, et al. (2005). Structural basis of West Nile virus neutralization by a therapeutic antibody. Nature 437(7059): 764-769.
O'Mullan, P., D. Craft, et al. (2009). Thrombin induces broad spectrum proteolysis in human serum samples. Clin Chem Lab Med 47(6): 685-693.
Op De Beeck, A., R. Molenkamp, et al. (2003). Role of the transmembrane domains of prM and E proteins in the formation of yellow fever virus envelope. J Virol 77(2): 813-820.
Parida, M., K. Horioke, et al. (2005). Rapid detection and differentiation of dengue virus serotypes by a real-time reverse transcription-loop-mediated isothermal amplification assay. J Clin Microbiol 43(6): 2895-2903.
Philips, M., A. G. Juul, et al. (1984). Human endothelial cells produce a plasminogen activator inhibitor and a tissue-type plasminogen activator-inhibitor complex. Biochim Biophys Acta 802(1): 99-110.
Randolph, V. B., G. Winkler, et al. (1990). Acidotropic amines inhibit proteolytic processing of flavivirus prM protein. Virology 174(2): 450-458.
Rico-Hesse, R. (1990). Molecular evolution and distribution of dengue viruses type 1 and 2 in nature. Virology 174(2): 479-493.
Russell, P. K. and A. Nisalak (1967). Dengue virus identification by the plaque reduction neutralization test. J Immunol 99(2): 291-296.
Shu, P. Y., S. F. Chang, et al. (2003). Development of group- and serotype-specific one-step SYBR green I-based real-time reverse transcription-PCR assay for dengue virus. J Clin Microbiol 41(6): 2408-2416.
Smith, T. J., W. E. Brandt, et al. (1970). Physical and biological properties of dengue-2 virus and associated antigens. J Virol 5(4): 524-532.
Steinitz, M., G. Klein, et al. (1977). EB virus-induced B lymphocyte cell lines producing specific antibody. Nature 269(5627): 420-422.
Steinitz, M. and S. Tamir (1982). Human monoclonal autoimmune antibody produced in vitro: rheumatoid factor generated by Epstein-Barr virus-transformed cell line. Eur J Immunol 12(2): 126-133.
Sutherland, M. R., H. M. Friedman, et al. (2004). Herpes simplex virus type 1-encoded glycoprotein C enhances coagulation factor VIIa activity on the virus. Thromb Haemost 92(5): 947-955.
Tee, H. P., S. H. How, et al. (2009). Risk factors associated with development of dengue haemorrhagic fever or dengue shock syndrome in adults in Hospital Tengku Ampuan Afzan Kuantan. Med J Malaysia 64(4): 316-320.
Tesh, R. B. (1979). A method for the isolation and identification of dengue viruses, using mosquito cell cultures. Am J Trop Med Hyg 28(6): 1053-1059.
Tricou, V., N. N. Minh, et al. (2011). Kinetics of Viremia and NS1 Antigenemia Are Shaped by Immune Status and Virus Serotype in Adults with Dengue. PLoS Negl Trop Dis 5(9): e1309.
Triplett, D. A. (2000). Coagulation and bleeding disorders: review and update. Clin Chem 46(8 Pt 2): 1260-1269.
Wang, S. M. and S. D. Sekaran (2010). Evaluation of a commercial SD dengue virus NS1 antigen capture enzyme-linked immunosorbent assay kit for early diagnosis of dengue virus infection. J Clin Microbiol 48(8): 2793-2797.
Watanabe, S., K. H. Tan, et al. (2012). The magnitude of dengue virus NS1 protein secretion is strain dependent and does not correlate with severe pathologies in the mouse infection model. J Virol 86(10): 5508-5514.
Wengler, G. (1991). The carboxy-terminal part of the NS 3 protein of the West Nile flavivirus can be isolated as a soluble protein after proteolytic cleavage and represents an RNA-stimulated NTPase. Virology 184(2): 707-715.
Winter, G., A. D. Griffiths, et al. (1994). Making antibodies by phage display technology. Annu Rev Immunol 12: 433-455.
Wu, Y. C. (1986). Epidemic dengue 2 on Liouchyou Shiang, Pingtung Country in 1981. Chinese J. Microbiol. Immunol. 19 (3): 203-211.
Yon, C., T. Teramoto, et al. (2005). Modulation of the nucleoside triphosphatase/RNA helicase and 5'-RNA triphosphatase activities of Dengue virus type 2 nonstructural protein 3 (NS3) by interaction with NS5, the RNA-dependent RNA polymerase. J Biol Chem 280(29): 27412-27419.
Young, P. R., P. A. Hilditch, et al. (2000). An antigen capture enzyme-linked immunosorbent assay reveals high levels of the dengue virus protein NS1 in the sera of infected patients. J Clin Microbiol 38(3): 1053-1057.
Yu, I. M., W. Zhang, et al. (2008). Structure of the immature dengue virus at low pH primes proteolytic maturation. Science 319(5871): 1834-1837.
Yuill, T. M., P. Sukhavachana, et al. (1968). Dengue-virus recovery by direct and delayed plaques in LLC-MK2 cells. Am J Trop Med Hyg 17(3): 441-448.
Zainah, S., A. H. Wahab, et al. (2009). Performance of a commercial rapid dengue NS1 antigen immunochromatography test with reference to dengue NS1 antigen-capture ELISA. J Virol Methods 155(2): 157-160.

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