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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 莊榮輝 | |
dc.contributor.author | Jie-Long He | en |
dc.contributor.author | 何杰龍 | zh_TW |
dc.date.accessioned | 2021-06-16T16:10:20Z | - |
dc.date.available | 2016-03-15 | |
dc.date.copyright | 2013-03-15 | |
dc.date.issued | 2013 | |
dc.date.submitted | 2013-02-25 | |
dc.identifier.citation | 林仕鈺 (2004) 1998-2003年台灣家禽流行性感冒病毒監測。行政院農委會家畜衛生試驗所禽流感資訊。
陳怡彤 (2009) H6亞型家禽流行性感冒病毒抗原及抗體酵素連結免疫吸附法之開發。碩士論文。台北:國立台灣大學獸醫學研究所。 陳建豪 (2006) 家禽流行性感冒病毒血球凝集素蛋白之表現及其應用。碩士論文。台北:國立台灣大學獸醫學研究所。 楊平政、黃元品、王金和 (2006) 屠宰場仿土雞血清之家禽流行性感冒流行病學調查分析。台灣獸醫誌 32 : 24-29。 劉怡君 (2010) 利用禽流感病毒 RNA 聚合酶 PA 和 PB2 之單株抗體探討其細胞感染機制。碩士論文。台北:國立台灣大學微生物與生化研究所。 鄭明珠、李敏旭、陳麗璇、劉玉彬、郭舒婷、李淑慧 (2007) 2006年野鳥家禽流行性感冒監測。家畜衛試所研報 42 : 43-50。 謝快樂、黃文正、沈瑞鴻、邱新育、李龍湖、呂榮修 (1992) 台灣地區家禽流行性感冒之研究 (Ⅲ) 雞群病毒株之分離同定和病原性試驗。台灣省畜牧獸醫學會會報 59 : 45-55。 Abe Y, Takashita E, Sugawara K, Matsuzaki Y, Muraki Y, et al. (2004) Effect of the addition of oligosaccharides on the biological activities and antigenicity of influenza A/H3N2 virus hemagglutinin. J Virol 78: 9605-9611. Alexander DJ (1982) Avian infuenza-recent developments. Bull World Health Organ 52: 341-359. Alexander DJ (2000) A review of avian influenza in different bird species. Vet Microbiol 74: 3-13. Almond JW (1977) A single gene determines the host range of influenza virus. Nature 270: 617-618. Baigent SJ, McCauley JW (2001) Glycosylation of haemagglutinin and stalk-length of neuraminidase combine to regulate the growth of avian influenza viruses in tissue culture. Virus Res 79: 177-185. Barbey-Martin C, Gigant B, Bizebard T, Calder LJ, Wharton SA, et al. (2002) An antibody that prevents the hemagglutinin low pH fusogenic transition. Virology 294: 70-74. Bizebard T, Gigant B, Rigolet P, Rasmussen B, Diat O, et al. (1995) Structure of influenza virus haemagglutinin complexed with a neutralizing antibody. Nature 376: 92-94. Brown EG (2000) Influenza virus genetics. Biomed Pharmacother 54: 196-209. Brownlee GG, Sharps JL (2002) The RNA polymerase of influenza a virus is stabilized by interaction with its viral RNA promoter. J Virol 76: 7103-7113. Carrat F, Flahault A (2007) Influenza vaccine: the challenge of antigenic drift. Vaccine 25: 6852-6862. Caton AJ, Brownlee GG, Yewdell JW, Gerhard W (1982) The antigenic structure of the influenza virus A/PR/8/34 hemagglutinin (H1 subtype). Cell 31: 417-427. Chanturiya AN, Basanez G, Schubert U, Henklein P, Yewdell JW, et al. (2004) PB1-F2, an influenza A virus-encoded proapoptotic mitochondrial protein, creates variably sized pores in planar lipid membranes. J Virol 78: 6304-6312. Chen GW, Yang CC, Tsao KC, Huang CG, Lee LA, et al. (2004) Influenza A virus PB1-F2 gene in recent Taiwanese isolates. Emerg Infect Dis 10: 630-636. Chen W, Calvo PA, Malide D, Gibbs J, Schubert U, et al. (2001) A novel influenza A virus mitochondrial protein that induces cell death. Nat Med 7: 1306-1312. Chen W, Zhong Y, Qin Y, Sun S, Li Z (2012) The evolutionary pattern of glycosylation sites in influenza virus (H5N1) hemagglutinin and neuraminidase. PloS One 7: e49224. Chen YT, Juang RH, He JL, Chu WY, Wang CH (2010) Detection of H6 influenza antibody by blocking enzyme-linked immunosorbent assay. Vet Microbiol 142: 205-210. Cheung TK, Poon LL (2007) Biology of influenza a virus. Ann N Y Acad Sci 1102: 1-25. Connor RJ, Kawaoka Y, Webster RG, Paulson JC (1994) Receptor specificity in human, avian, and equine H2 and H3 influenza virus isolates. Virology 205: 17-23. Corti D, Voss J, Gamblin SJ, Codoni G, Macagno A, et al. (2011) A neutralizing antibody selected from plasma cells that binds to group 1 and group 2 influenza A hemagglutinins. Science 333: 850-856. Couceiro JN, Paulson JC, Baum LG (1993) Influenza virus strains selectively recognize sialyloligosaccharides on human respiratory epithelium; the role of the host cell in selection of hemagglutinin receptor specificity. Virus Res 29: 155-165. Cox NJ, Subbarao K (2000) Global epidemiology of influenza: past and present. Annu Rev Med 51: 407-421. Daniels R, Kurowski B, Johnson AE, Hebert DN (2003) N-linked glycans direct the cotranslational folding pathway of influenza hemagglutinin. Mol Cell 11: 79-90. Deshpande KL, Fried VA, Ando M, Webster RG (1987) Glycosylation affects cleavage of an H5N2 influenza virus hemagglutinin and regulates virulence. Proc Natl Acad Sci U S A 84: 36-40. Easterday BC, Hinshaw VS, Halvorson DA(1997) Influenza. In: Diseases of poultry. 10th ed B W Calnek, H J Barnes, C W Beard, L R Mcdougald, and Y M Saif, eds Iowa State University Press, Ames, Iowa: 583-605. Ekiert DC, Friesen RH, Bhabha G, Kwaks T, Jongeneelen M, et al. (2011) A highly conserved neutralizing epitope on group 2 influenza A viruses. Science 333: 843-850. Ekiert DC, Kashyap AK, Steel J, Rubrum A, Bhabha G, et al. (2012) Cross-neutralization of influenza A viruses mediated by a single antibody loop. Nature 489: 526-532. Gibbs JS, Malide D, Hornung F, Bennink JR, Yewdell JW (2003) The influenza A virus PB1-F2 protein targets the inner mitochondrial membrane via a predicted basic amphipathic helix that disrupts mitochondrial function. J Virol 77: 7214-7224. He JL, Hsieh MS, Chiu YC, Juang RH, Wang CH (2013) Preparation of monoclonal antibodies against poor immunogenic avian influenza virus proteins. J Immunol Methods 387: 43-50. Horimoto T, Kawaoka Y (1995) Direct reverse transcriptase PCR to determine virulence potential of influenza A viruses in birds. J Clin Microbiol 33: 748-751. Hsu CN, Wang CH (2006) Sequence comparison between two quasi strains of H6N1 with different pathogenicity from a single parental isolate. J Microbiol Immunol Infect 39: 292-296. Huang K, Bahl J, Fan XH, Vijaykrishna D, Cheung CL, et al. (2010) Establishment of an H6N2 influenza virus lineage in domestic ducks in southern China. J Virol 84: 6978-6986. Inglis SC, Barrett T, Brown CM, Almond JW (1979) The smallest genome RNA segment of influenza virus contains two genes that may overlap. Proc Natl Acad Sci U S A 76: 3790-3794. Inglis SC, Brown CM (1981) Spliced and unspliced RNAs encoded by virion RNA segment 7 of influenza virus. Nucleic Acids Res 9: 2727-2740. Katz JM, Lu X, Tumpey TM, Smith CB, Shaw MW, et al. (2000) Molecular correlates of influenza A H5N1 virus pathogenesis in mice. J Virol 74: 10807-10810. Kawaoka Y, Naeve CW, Webster RG (1984) Is virulence of H5N2 influenza viruses in chickens associated with loss of carbohydrate from the hemagglutinin? Virology 139: 303-316. Kawaoka Y, Webster RG (1989) Interplay between carbohydrate in the stalk and the length of the connecting peptide determines the cleavability of influenza virus hemagglutinin. J Virol 63: 3296-3300. Kida H, Ito T, Yasuda J, Shimizu Y, Itakura C, et al. (1994) Potential for transmission of avian influenza viruses to pigs. J Gen Virol 75 ( Pt 9): 2183-2188. Kido H, Okumura Y, Yamada H, Le TQ, Yano M (2007) Proteases essential for human influenza virus entry into cells and their inhibitors as potential therapeutic agents. Curr Pharm Des 13: 405-414. Kilbourne ED (1987) Influenza. Plenum medical book company. New York and London. Kilbourne ED, Cerini CP, Khan MW, Mitchell JW, Jr., Ogra PL (1987) Immunologic response to the influenza virus neuraminidase is influenced by prior experience with the associated viral hemagglutinin. I. Studies in human vaccinees. J Immunol 138: 3010-3013. Klenk HD, Garten W (1994) Host cell proteases controlling virus pathogenicity. Trends Microbiol 2: 39-43. Klenk HD, Rott R, Orlich M, Blodorn J (1975) Activation of influenza A viruses by trypsin treatment. Virology 68: 426-439. Klenk HD, Wagner R, Heuer D, Wolff T (2002) Importance of hemagglutinin glycosylation for the biological functions of influenza virus. Virus Res 82: 73-75. Knossow M, Skehel JJ (2006) Variation and infectivity neutralization in influenza. Immunology 119: 1-7. Kohler G, Milstein C (1975) Continuous cultures of fused cells secreting antibody of predefined specificity. Nature 256: 495-497. Lamb RA, Krug RM (2001) Orthomyxoviridae: the viruses and their replication. In: D.M. Knipe and P.M. Howley (Eds), Fields Virology, Vol. 1, Lippincott Williams and Wilkins, Philadelphia., pp. 1487-1531. Laver WG, Air GM, Webster RG (1981) Mechanism of antigenic drift in influenza virus. Amino acid sequence changes in an antigenically active region of Hong Kong (H3N2) influenza virus hemagglutinin. J Mol Biol 145: 339-361. Lazarowitz SG, Choppin PW (1975) Enhancement of the infectivity of influenza A and B viruses by proteolytic cleavage of the hemagglutinin polypeptide. Virology 68: 440-454. Lee MS, Chang PC, Shien JH, Cheng MC, Chen CL, et al. (2006) Genetic and pathogenic characterization of H6N1 avian influenza viruses isolated in Taiwan between 1972 and 2005. Avian Dis 50: 561-571. Lee MS, Chang PC, Shien JH, Cheng MC, Shieh HK (2001) Identification and subtyping of avian influenza viruses by reverse transcription-PCR. J Virol Methods 97: 13-22. Lee PS, Yoshida R, Ekiert DC, Sakai N, Suzuki Y, et al. (2012) Heterosubtypic antibody recognition of the influenza virus hemagglutinin receptor binding site enhanced by avidity. Proc Natl Acad Sci U S A 109: 17040-17045. Lentz MR, Webster RG, Air GM (1987) Site-directed mutation of the active site of influenza neuraminidase and implications for the catalytic mechanism. Biochemistry 26: 5351-5358. Lonsdale R, Pau MG, Oerlemans M, Ophorst C, Vooys A, et al. (2003) A rapid method for immunotitration of influenza viruses using flow cytometry. J Virol Methods 110: 67-71. Lupiani B, Reddy SM (2009) The history of avian influenza. Comp Immunol Microbiol Infect Dis 32: 311-323. Martin K, Helenius A (1991) Nuclear transport of influenza virus ribonucleoproteins: the viral matrix protein (M1) promotes export and inhibits import. Cell 67: 117-130. Matrosovich M, Zhou N, Kawaoka Y, Webster R (1999) The surface glycoproteins of H5 influenza viruses isolated from humans, chickens, and wild aquatic birds have distinguishable properties. J Virol 73: 1146-1155. Matrosovich MN, Matrosovich TY, Gray T, Roberts NA, Klenk HD (2004) Human and avian influenza viruses target different cell types in cultures of human airway epithelium. Proc Natl Acad Sci U S A 101: 4620-4624. Murphy BR, Webster RG (1996) Orthomyxoviruses. In fields virology, 3rd ed B N Fields, D M Knipe, P M Ho wley, R M Chanock, J L Melnick, T P Monath, B Roizman, and S T Straus, eds Lippincott-Raven Press, Philadelphia., pp. 1397-1445. Murti KG, Bean WJ, Jr., Webster RG (1980) Helical ribonucleoproteins of influenza virus: an electron microscopic analysis. Virology 104: 224-229. Neumann G, Kawaoka Y (2006) Host range restriction and pathogenicity in the context of influenza pandemic. Emerg Infect Dis 12: 881-886. Nobusawa E, Aoyama T, Kato H, Suzuki Y, Tateno Y, et al. (1991) Comparison of complete amino acid sequences and receptor-binding properties among 13 serotypes of hemagglutinins of influenza A viruses. Virology 182: 475-485. OIE (2009) Update on highly pathogenic avian influenza in animals (type H5 and H7). Avian influenza. http://www.oie.int/downld/AVIAN%20INFLUENZA/A_AI-Asia.htm. OIE. (2008) World Organization for Animal Health. Avian influenza. In: Manual of diagnostic tests and vaccines for terrestrial animals. http://www.oie.int/eng/ normes/mmanual/A_00037.htm. O'Neill RE, Jaskunas R, Blobel G, Palese P, Moroianu J (1995) Nuclear import of influenza virus RNA can be mediated by viral nucleoprotein and transport factors required for protein import. J Biol Chem 270: 22701-22704. O'Neill RE, Talon J, Palese P (1998) The influenza virus NEP (NS2 protein) mediates the nuclear export of viral ribonucleoproteins. EMBO J 17: 288-296. Perdue ML, Garcia M, Senne D, Fraire M (1997) Virulence-associated sequence duplication at the hemagglutinin cleavage site of avian influenza viruses. Virus Res 49: 173-186. Philpott M, Easterday BC, Hinshaw VS (1989) Neutralizing epitopes of the H5 hemagglutinin from a virulent avian influenza virus and their relationship to pathogenicity. J Virol 63: 3453-3458. Philpott M, Hioe C, Sheerar M, Hinshaw VS (1990) Hemagglutinin mutations related to attenuation and altered cell tropism of a virulent avian influenza A virus. J Virol 64: 2941-2947. Privalsky ML, Penhoet EE (1978) Influenza virus proteins: identity, synthesis, and modification analyzed by two-dimensional gel electrophoresis. Proc Natl Acad Sci U S A 75: 3625-3629. Privalsky ML, Penhoet EE (1981) The structure and synthesis of influenza virus phosphoproteins. J Biol Chem 256: 5368-5376. Reed LJ, Muench H (1938) A simple method of estimating fifty percent endpoints. The American Journal of Hygiene 27: 493–497. Richardson JC, Akkina RK (1991) NS2 protein of influenza virus is found in purified virus and phosphorylated in infected cells. Arch Virol 116: 69-80. Rimmelzwaan GF, Baars M, Claas EC, Osterhaus AD (1998) Comparison of RNA hybridization, hemagglutination assay, titration of infectious virus and immunofluorescence as methods for monitoring influenza virus replication in vitro. J Virol Methods 74: 57-66. Rogers GN, Paulson JC, Daniels RS, Skehel JJ, Wilson IA, et al. (1983) Single amino acid substitutions in influenza haemagglutinin change receptor binding specificity. Nature 304: 76-78. Rott R (1992) The pathogenic determinant of influenza virus. Vet Microbiol 33: 303-310. Scholtissek C (1995) Molecular evolution of influenza viruses. Virus Genes 11: 209-215. Schulze-Horsel J, Genzel Y, Reichl U (2008) Flow cytometric monitoring of influenza A virus infection in MDCK cells during vaccine production. BMC Biotechnol 8: 45. Shinya K, Ebina M, Yamada S, Ono M, Kasai N, et al. (2006) Avian flu: influenza virus receptors in the human airway. Nature 440: 435-436. Skehel JJ, Stevens DJ, Daniels RS, Douglas AR, Knossow M, et al. (1984) A carbohydrate side chain on hemagglutinins of Hong Kong influenza viruses inhibits recognition by a monoclonal antibody. Proc Natl Acad Sci U S A 81: 1779-1783. Skehel JJ, Wiley DC (2000) Receptor binding and membrane fusion in virus entry: the influenza hemagglutinin. Annu Rev Biochem 69: 531-569. Stech O, Veits J, Weber S, Deckers D, Schroer D, et al. (2009) Acquisition of a polybasic hemagglutinin cleavage site by a low-pathogenic avian influenza virus is not sufficient for immediate transformation into a highly pathogenic strain. J Virol 83: 5864-5868. Stieneke-Grober A, Vey M, Angliker H, Shaw E, Thomas G, et al. (1992) Influenza virus hemagglutinin with multibasic cleavage site is activated by furin, a subtilisin-like endoprotease. EMBO J 11: 2407-2414. Subbarao K, Klimov A, Katz J, Regnery H, Lim W, et al. (1998) Characterization of an avian influenza A (H5N1) virus isolated from a child with a fatal respiratory illness. Science 279: 393-396. Sun S, Wang Q, Zhao F, Chen W, Li Z (2011) Glycosylation site alteration in the evolution of influenza A (H1N1) viruses. PloS One 6: e22844. Suzuki Y, Ito T, Suzuki T, Holland RE, Jr., Chambers TM, et al. (2000) Sialic acid species as a determinant of the host range of influenza A viruses. J Virol 74: 11825-11831. Swayne DE (2008) The global nature of avian influenza, IA. In: D.E. Swayne (Ed), Avian influenza, Blackwell, Ames. Taylor HP, Armstrong SJ, Dimmock NJ (1987) Quantitative relationships between an influenza virus and neutralizing antibody. Virology 159: 288-298. Temoltzin-Palacios F, Thomas DB (1994) Modulation of immunodominant sites in influenza hemagglutinin compromise antigenic variation and select receptor-binding variant viruses. J Exp Med 179: 1719-1724. Thanh TT, van Doorn HR, de Jong MD (2008) Human H5N1 influenza: current insight into pathogenesis. Int J Biochem Cell Biol 40: 2671-2674. Tong S, Li Y, Rivailler P, Conrardy C, Castillo DA, et al. (2012) A distinct lineage of influenza A virus from bats. Proc Natl Acad Sci U S A 109: 4269-4274. Tsuchiya E, Sugawara K, Hongo S, Matsuzaki Y, Muraki Y, et al. (2001) Antigenic structure of the haemagglutinin of human influenza A/H2N2 virus. J Gen Virol 82: 2475-2484. Turek R, Gresikova M, Tumova B (1984) Isolation of influenza A virus and paramyxoviruses from sentinel domestic ducks. Acta Virol 28: 156-158. Vigerust DJ, Shepherd VL (2007) Virus glycosylation: role in virulence and immune interactions. Trends Microbiol 15: 211-218. Vigerust DJ, Ulett KB, Boyd KL, Madsen J, Hawgood S, et al. (2007) N-linked glycosylation attenuates H3N2 influenza viruses. J Virol 81: 8593-8600. Vines A, Wells K, Matrosovich M, Castrucci MR, Ito T, et al. (1998) The role of influenza A virus hemagglutinin residues 226 and 228 in receptor specificity and host range restriction. J Virol 72: 7626-7631. Wang CC, Chen JR, Tseng YC, Hsu CH, Hung YF, et al. (2009) Glycans on influenza hemagglutinin affect receptor binding and immune response. Proc Natl Acad Sci U S A 106: 18137-18142. Wang W, Lu B, Zhou H, Suguitan AL, Jr., Cheng X, et al. (2010) Glycosylation at 158N of the hemagglutinin protein and receptor binding specificity synergistically affect the antigenicity and immunogenicity of a live attenuated H5N1 A/Vietnam/1203/2004 vaccine virus in ferrets. J Virol 84: 6570-6577. Wanzeck K, Boyd KL, McCullers JA (2011) Glycan shielding of the influenza virus hemagglutinin contributes to immunopathology in mice. Am J Respir Crit Care Med 183: 767-773. Webster RG, Bean WJ, Gorman OT, Chambers TM, Kawaoka Y (1992) Evolution and ecology of influenza A viruses. Microbiol Rev 56: 152-179. Webster RG, Laver WG (1980) Determination of the number of nonoverlapping antigenic areas on Hong Kong (H3N2) influenza virus hemagglutinin with monoclonal antibodies and the selection of variants with potential epidemiological significance. Virology 104: 139-148. Webster RG, Rott R (1987) Influenza virus A pathogenicity: the pivotal role of hemagglutinin. Cell 50: 665-666. White J, Matlin K, Helenius A (1981) Cell fusion by Semliki Forest, influenza, and vesicular stomatitis viruses. J Cell Biol 89: 674-679. Whittle JR, Zhang R, Khurana S, King LR, Manischewitz J, et al. (2011) Broadly neutralizing human antibody that recognizes the receptor-binding pocket of influenza virus hemagglutinin. Proc Natl Acad Sci U S A 108: 14216-14221. Wiley DC, Wilson IA, Skehel JJ (1981) Structural identification of the antibody-binding sites of Hong Kong influenza haemagglutinin and their involvement in antigenic variation. Nature 289: 373-378. Wilson IA, Skehel JJ, Wiley DC (1981) Structure of the haemagglutinin membrane glycoprotein of influenza virus at 3 A resolution. Nature 289: 366-373. Wrammert J, Koutsonanos D, Li GM, Edupuganti S, Sui J, et al. (2011) Broadly cross-reactive antibodies dominate the human B cell response against 2009 pandemic H1N1 influenza virus infection. J Exp Med 208: 181-193. Wu ZL, Ethen C, Hickey GE, Jiang W (2009) Active 1918 pandemic flu viral neuraminidase has distinct N-glycan profile and is resistant to trypsin digestion. Biochem Biophys Res Commun 379: 749-753. Xu R, Ekiert DC, Krause JC, Hai R, Crowe JE, Jr., et al. (2010) Structural basis of preexisting immunity to the 2009 H1N1 pandemic influenza virus. Science 328: 357-360. Yasuda J, Nakada S, Kato A, Toyoda T, Ishihama A (1993) Molecular assembly of influenza virus: association of the NS2 protein with virion matrix. Virology 196: 249-255. Ye ZP, Baylor NW, Wagner RR (1989) Transcription-inhibition and RNA-binding domains of influenza A virus matrix protein mapped with anti-idiotypic antibodies and synthetic peptides. J Virol 63: 3586-3594. Yoshida R, Igarashi M, Ozaki H, Kishida N, Tomabechi D, et al. (2009) Cross-protective potential of a novel monoclonal antibody directed against antigenic site B of the hemagglutinin of influenza A viruses. PLoS Pathog 5: e1000350. Zambon MC (1999) Epidemiology and pathogenesis of influenza. J Antimicrob Chemother 44 Suppl B: 3-9. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/62784 | - |
dc.description.abstract | 本研究以病毒顆粒與合成胜肽為抗原進行小鼠免疫,生產專一性的單株抗體,並匯集成「禽流感病毒蛋白質體單株抗體庫」。我們已經篩選出一些單株抗體,可以辨識 H6N1 亞型 A/chicken/Taiwan/2838V/00 (2838V) 低病原性禽流感病毒的主要結構蛋白質,其中包括可專一性辨識 H6 亞型病毒血球凝集素次體 1 的單株抗體 (EB2 mAb),被用於追蹤受 2838V 病毒感染細胞表面 HA1 以研究醣類對感染機制的交互作用。HA 活化切位 (HACS) 是決定禽流感病毒致病力的重要因子,低病原性禽流感在此活化切位上只有單一的鹼性胺基酸,僅能被 trypsin 給切開。但是除了活化切位外,HA1 上還有其他的切位可能被 trypsin 攻擊,這些切位如何抵抗 trypsin 的作用成為本研究關注的焦點。觀察切醣後經 trypsin 處理的病毒樣本,本研究發現 2838V 病毒 HA1 上的 N167 醣類可能保護 R201 切位抵抗 trypsin 切割。當這個醣類被切除,2838V HA1 便容易被 trypsin 切開。由細胞 in vitro 與雞胚蛋 in vivo 的實驗證實,2838V 同時切醣與外加 trypsin 會降低感染能力,這代表受 N167 醣類保護抵抗 trypsin 切割的 R201 切位附近的區域對於調控 2838V 病毒感染扮演重要角色。此外,本研究亦發現 EB2 單株抗體辨認 2838V HA 頭部接近受體結合區的抗原決定位。此抗體對於 H6 亞型病毒的中和能力已經流式細胞儀偵測感染細胞與雞胚蛋實驗確認。以 EB2 單株抗體辨認的抗原決定位序列合成的胜肽對小鼠進行免疫注射,小鼠抗血清具有與 EB2 單株抗體相似的中和反應能力。此抗原決定位可能具有臨床應用的價值,做為禽流感 HA 蛋白質結構與功能性研究的工具,甚至協助 HA 次單元疫苗的開發。 | zh_TW |
dc.description.abstract | In this study, a monoclonal antibody bank (mAb bank) against A/chicken/Taiwan/2838V/00 (2838V) virus was established by either the viron or synthetic peptides as the antigen. Several mAb against the major virion proteins of 2838V have been screened including a high specific hemagglutinin 1 subunit mAb (EB2) recognizing H6 subtype AIV. This mAb was used to trace the HA1 variation by infecting cells, and to investigate the glycan interaction in HA1. HACS is critical to viral pathogenicity. Low pathogenic AIV contains an HA cleavage site (HACS) having only one arginine or lysine which could be cleaved by trypsin. In addition, many cleavage sites excluding HACS are located in HA1. How the other cleavage sites resist trypsin digestion is our great interest. In the present study, our observations indicated that the glycosylation at N167 of the 2838V HA1 could protect R201 site from tryptic cleavage. The 2838V HA1 became sensitive to tryptic cleavage while the N167 glycans were removed. The infectivity of 2838V was also decreased upon pre-treated it with PNGase-F and trypsin in vitro and in vivo assay. Our observations suggest that the inaccessibility of the R201 residue of HA1 for tryptic cleavage, which is sterically hindered by glycosylation at N167, is an important factor in determining the infectivity of the 2838V AIV. Furthermore, we found that the EB2 mAb recognizes a linear HA epitope at the globular head near the receptor binding site of the 2838V. Flow cytometry of AIV-infected cells and embryonated eggs assay suggested that the neutralizing activity of this mAb for H6 AIVs. The mice injection with the EB2-defined epitope peptide showed similarly neutralizing activity to EB2 mAb. This epitope might be useful for clinical applications and as a tool for further study of the structure and function of the AIV HA protein, even subunit vaccine development. | en |
dc.description.provenance | Made available in DSpace on 2021-06-16T16:10:20Z (GMT). No. of bitstreams: 1 ntu-102-D93b47202-1.pdf: 11716997 bytes, checksum: 155f5bdef9e7e37689ab300a7c4e2df6 (MD5) Previous issue date: 2013 | en |
dc.description.tableofcontents | 口試委員會審定書 I
誌謝 II 中文摘要 III 英文摘要 IV 目錄 V 圖表目錄 VIII 第 一 章 緒論 1 1.1 禽流感研究的歷史背景 1 1.2 禽流感病毒簡介 4 1.2.1 病毒的分類與命名規則 4 1.2.2 病毒形態與構造 5 1.2.3 病毒的基因體 7 1.2.4 病毒的複製 8 1.2.5 病毒的突變 10 1.2.6 病毒的蛋白質體 10 1.3 血球凝集素是影響禽流感病毒毒力的關鍵 16 1.3.1 與宿主受體表面醣類的親和力決定病毒專一性 16 1.3.2 蛋白酶活化切位影響病毒毒力 17 1.3.3 病毒蛋白質醣基化所扮演的角色 19 1.4 禽流感病毒研究方法 23 1.4.1 病毒增殖 23 1.4.2 測定病毒效價 24 1.4.3 以單株抗體進行蛋白質體學研究與應用 24 1.4.4 禽流感病毒蛋白質體單株抗體庫 26 1.4.5 篩選具中和反應的單株抗體 27 1.4.6 研究醣蛋白質的工具 28 1.4.7 以流式細胞儀評估低病原性禽流感病毒的感染能力 29 1.5研究動機及目的 30 第 二 章 材料與方法 31 2.1 病毒樣本處理 31 2.2 建立 H6N1 亞型禽流感病毒蛋白質體單株抗體庫 31 2.2.1 第一階段:以病毒顆粒為抗原 31 2.2.2 第二階段:設計短鏈胜肽為抗原 32 2.2.3 分析單株抗體辨識的抗原身分 34 2.2.4 雙重篩選法確定單株抗體的專一性 34 2.2.5 鑑定單株抗體型別 34 2.2.6 分析單株抗體的抗原決定位 35 2.2.7 兔的傳統抗血清製備與特性檢測 36 2.2.8 禽流感病毒 H5N2 亞型傳統抗血清製備 37 2.3 製備 2838V HA1 重組蛋白質 38 2.3.1 構築 2838V HA1 部分片段重組蛋白質之重組質體 38 2.3.2 重組蛋白質之表現與純化 40 2.4 禽流感病毒蛋白質體分析 40 2.4.1 比較各禽流感病毒株之二維電泳圖譜差異 40 2.4.2 以 LC-MS/MS 鑑定禽流感病毒二維電泳圖譜之主要蛋白質點 41 2.5 禽流感病毒蛋白質序列分析 41 2.5.1 禽流感病毒蛋白質序列分析與資料庫比對 41 2.5.2 禽流感病毒主要蛋白質分子模擬 41 2.6 分析禽流感病毒 HA1 的醣類與 trypsin 切位之關係 42 2.6.1 以酵素去除禽流感病毒蛋白質 N-linked 醣類 42 2.6.2 以凝集素偵測各禽流感病毒醣蛋白質醣類的差異 42 2.6.3 以醣晶片檢測禽流感病毒醣蛋白質醣類的差異 45 2.6.4 分析禽流感病毒 HA1 的 trypsin 切位 46 2.6.5 禽流感病毒 HA1 對 trypsin 的耐受性與醣類的關係 46 2.6.6 預測禽流感病毒 HA1 上醣基化位置 46 2.6.7 以 LC-MS/MS 追蹤受醣類保護的 trypsin 切割位 47 2.6.8 禽流感病毒 2838V HA1 與 trypsin 的分子對接 47 2.7 以流式細胞儀檢測禽流感病毒 47 2.7.1 禽流感病毒感染 DF1 細胞 47 2.7.2 禽流感病毒細胞感染半定量測定 48 2.7.3 以流式細胞儀測定禽流感病毒感染細胞能力 48 2.7.4 以 RT-PCR 確認流式細胞儀檢測的結果 49 2.7.5 評估醣類對 trypsin 耐受性與病毒感染細胞能力的影響 50 2.7.6 評估單株抗體的中和反應能力 50 2.8 以雞胚蛋檢測禽流感病毒的感染力 50 2.8.1 各禽流感病毒株的雞胚蛋感染半致死測定 50 2.8.2 以雞胚蛋評估醣類對 trypsin 耐受性與病毒感染力的影響 50 2.8.3 以雞胚蛋評估 EB2 單株抗體的中和反應能力 51 2.9 可誘導中和反應的 HA1 抗原決定位 51 2.9.1 以 EB2 單株抗體辨認的抗原決定位合成胜肽免疫小鼠 51 2.9.2 以雞胚蛋評估小鼠抗血清的中和反應能力 51 2.9.3 單株抗體辨認的抗原決定位與資料庫序列比對 52 第 三 章 結果與討論 53 3.1 比較不同禽流感病毒株的蛋白質體 53 3.2 比較不同禽流感病毒株的二維膠體電泳蛋白質圖譜 53 3.3 建立 H6N1 亞型禽流感病毒蛋白質體抗體庫 57 3.3.1 以病毒樣本為抗原製備 HA 的單株抗體 57 3.3.2 禽流感病毒 H6 亞型 HA1 單株抗體的特性 58 3.4 禽流感病毒 2838V HA 的醣類末端仍為禽類的 α-2,3 SAL 66 3.5 切醣酵素可切除禽流感病毒 HA 的 N-linked 醣類 67 3.6 禽流感病毒 2838V HA1 的醣類可保護蛋白質抵抗蛋白酶切割 72 3.7 醣類保護 HA1 抵抗蛋白酶切割只發生於 2838V 病毒株 77 3.8 禽流感病毒 3233 HA1 的 N167 醣基化位置缺失 81 3.9 以 LC-MS/MS 追蹤醣類保護 2838V HA1 的可能區域 82 3.10 分析 EB2 單株抗體的抗原決定位 85 3.11 提出「HA 頭部醣類遮蔽切位以抵抗 trypsin 切割」假說 87 3.12 以流式細胞儀結合 EB2 單株抗體評估禽流感病毒的感染率 89 3.13 醣類遮蔽切位可保護 2838V 病毒感染細胞的能力 93 3.14 醣類遮蔽切位可保護 2838V 病毒感染雞胚蛋的能力 95 3.15 以細胞證實 EB2 是有效的中和抗體 98 3.16 以雞胚蛋證實 EB2 是有效的中和抗體 100 3.17 評估 EB2 單株抗體的抗原決定位誘導小鼠免疫反應 102 3.18 資料庫比對顯示 EB2 單株抗體可涵蓋大部分 H6 亞型病毒 104 3.19 高產能篩選禽流感病毒低免疫性蛋白質單株抗體的新策略 105 第 四 章 總結 109 參考文獻 111 附錄 119 A.1一般蛋白質分析 119 A.2二維膠體電泳 127 A.3單株抗體之製備 131 A.4蛋白質體學技術 144 A.5病毒樣本處理 146 A.6製備重組蛋白質 151 | |
dc.language.iso | zh-TW | |
dc.title | 血球凝集素上的醣基調控 H6N1 禽流感病毒感染 | zh_TW |
dc.title | The glycan of hemagglutinin regulates the H6N1 avian influenza virus infection. | en |
dc.type | Thesis | |
dc.date.schoolyear | 101-1 | |
dc.description.degree | 博士 | |
dc.contributor.coadvisor | 王金和 | |
dc.contributor.oralexamcommittee | 張明富,張世宗,陳翰民 | |
dc.subject.keyword | 禽流感病毒,單株抗體,活化切割位,醣類交互作用, | zh_TW |
dc.subject.keyword | Avian influenza virus,monoclonal antibody,HA cleavage site,glycan interaction, | en |
dc.relation.page | 161 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2013-02-26 | |
dc.contributor.author-college | 生命科學院 | zh_TW |
dc.contributor.author-dept | 生化科技學系 | zh_TW |
顯示於系所單位: | 生化科技學系 |
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