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研究生:黃志傑
研究生(外文):Ji-Jia Huang
論文名稱:日本血吸蟲26KDaGST疫苗抗原被動免疫分析及應用鋁佐劑誘導免疫效應之研究
論文名稱(外文):The studies of passive immunity on Schistosoma japonicum by 26 KDa GST and effects of Alum adjuvant on immune response
指導教授:李金木李金木引用關係
指導教授(外文):Kin-Mu Lee
學位類別:碩士
校院名稱:國立陽明大學
系所名稱:寄生蟲學研究所
學門:醫藥衛生學門
學類:醫學學類
論文種類:學術論文
論文出版年:2001
畢業學年度:89
語文別:中文
論文頁數:62
中文關鍵詞:日本血吸蟲
外文關鍵詞:Schistosoma japonicum
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中 文 摘 要
血吸蟲為世界衛生組織列為第二重大之熱帶寄生蟲疾病,三種血吸蟲以日本血吸蟲之致病力最嚴重。日本血吸蟲26KDa 麩氨基硫S-轉移(glutathione S-transferase;GST)已經被認定為一良好的疫苗候選者,本實驗室曾運用RT-PCR技術將台灣株日本血吸蟲26KDa GST的cDNA核酸選殖出來,於大腸桿菌中表現出重組蛋白,實驗證明其具疫苗保護效果。本研究論文進一步探討該疫苗抗原對於不同地域株日本血吸蟲感染預防之適用性。實驗結果顯示,數個不同地域株之日本血吸蟲26KDa GST的cDNA核酸序列極為相似,且所編碼之氨基酸序列完全相同。台灣及中國湖北株日本血吸蟲天然及重組蛋白免疫小鼠後所獲得之減蟲率及交叉保護力相似,顯示此26KDa GST疫苗抗原將可廣泛適用於各個不同地域株。以台灣株日本血吸蟲26KDa GST多源抗體及單株抗體進行被動免疫,分別得到50.2﹪及40﹪之減蟲率,證明本疫苗所誘發之保護效果與體液性免疫具有絕對的關聯。實驗結果亦顯示,弗氏佐劑或鋁佐劑混合26KDa GST免疫誘發之抗體種類不同,應用鋁佐劑未能獲得良好減蟲效果,小鼠血清中所存在之保護性抗體可能與IgG2b有關。此結果顯示疫苗免疫使用之佐劑種類是影響保護效果及誘發不同抗體類別之重要因素。

Abstract
Schistosomiasis is listed as the second most important tropical parasitic disease by World Health Organization. Among the three major human schistosomes, Schistosoma japonicum is the most pathogenic. The cDNA of 26KDa GST from the total RNA of adult worm of a Taiwanese strain of S. japonicum was synthesized and amplified by using RT-PCR technique, followed by the subcloning in our laboratory. The recombinant proteins were then expressed in Escherichia coli and demonstrated to be a good candidate vaccine. In this studies , the suitability of using 26KDa GST as a vaccine candidate against different strains of S. japonicum was evaluated. The results showed that the cDNA sequences of 26KDa GST among different strain of S. japonicum are similar and encode the same amino acid sequences. The worm reduction rate and cross protective immunity in mice immunized with native and recombinant GST26 from Taiwanese and Hupei strain of the parasite had similar results, indicating that the 26KDa GST can be a vaccine condidate against the different strains of S. japonicum. Passive transfer experiment using sera from the mice immunized with recombinant GST26, as well as monoclonal antibodies, showed a worm reduction rate of 50.2% and 40% respectively, indicating the close relationship between the protective immunity induced by GST26 and antibodies. The level of anti 26KDa GST IgG2b antibody in immunized mice with Freund's adjuvant was significantly higher than in the mice using Alum adjuvant. Also, poor reductive rate in mice group was obtained from the Alum adjuvant. This results showed that different adjuvant combined with subunit vaccine might produce different subclasses of antibodies which will lead to different protective effect.

中文摘要………………………………………………………….1
英文摘要………………………………………………………….2
第一章 緒論…………………………………………………… 3
第二章 材料與方法…………………………………………...11
實驗動物………………………………………………………11
生活史維持……………………………………………………11
SjCH及SjCA 26 KDa GST cDNA基因定序分析…………...12
SjT、SjCH 26 KDa rGST之製備…….………………………13
SjCH nGSTs(26~28 KDa)蛋白之製備....................................15
SjT、SjCH 26 KDa rGST與SjCH nGSTs免疫小鼠對SjT及SjCH感染抵抗能力之比較......................................................16
連脢免疫吸附分析(ELISA)...................................................16
西方墨點法(Western blotting)….............................................17
SjT 26 KDa rGST免疫抗血清的取得......................................18
SjT 26 KDa rGST免疫抗血清被動轉移及交叉被動保護實驗………………………………………………………………18
日本血吸蟲26 KDa GST蛋白單株抗體之取得……………..19
單株抗體的被動免疫保護力實驗……………………………20
SjT 26 KDa rGST混合弗氏佐劑或鋁佐劑之免疫效果與抗體誘發種類及效價評估…………………………………………20
統計分析..…………………………………………………….21
第三章 結果………………………………………………..…22
日本血吸蟲26 KDa GST cDNA基因定序分析………………………………………………………………22
SjT、SjCH 26 KDa rGST及SjCH nGSTs(26~28 KDa)之製備………………………………………………………………22
SjT、SjCH 26 KDa rGST與SjCH nGSTs免疫小鼠對SjT及SjCH感染抵抗能力之比較................................................…. 24
SjT 26 KDa rGST免疫抗血清之取得與免疫抗血清被動轉移及交叉被動保護實驗.........................................................…..24
日本血吸蟲26 KDa GST蛋白單株抗體的被動免疫保護力實驗......................................................................................….....25
SjT 26 KDa rGST混合弗氏佐劑或鋁佐劑之免疫效果與抗體誘發種類及效價評估...............................................................26
第四章 討論…………………………………………………..28
致 謝…………………………………………………………..35
參考文獻......................................................................................36
圖表及說明..................................................................................46

羅進宗. 1990. 彰化縣日本血吸蟲疫區住民、學童及中國探親榮民之寄生蟲調查。中華民國寄生蟲學雜誌 3: 97-100.
周述龍及林建銀主編. 1989. 血吸蟲學,第一版。科學出版社。
鄭江及屈振麒. 1993. 全國血吸蟲病檢查總結。 中國血吸蟲病流行狀況-1989年全國抽樣調查。 高淑芬主編。 成都科技大學出版社。 pp. 16-30.
曾憲芳及余炳楨. 1994. 血吸蟲(裂體吸蟲)。 人體寄生蟲學,第二版。 趙慰先主編。 人民衛生出版社。 pp. 310-329.
趙慰先及高淑芬主編. 1996. 實用血吸蟲病學,第一版。人民衛生出版社。
柳舒祥. 1996. 台灣株日本血吸蟲26 KDa麩氨基硫S-轉移基因之分子選殖及疫苗特性之研究. 碩士論文.
楊明芬. 1999. 以IL-12作為26 KDa麩氨基硫S-轉移疫苗佐劑抗日本血吸蟲之免疫效應. 碩士論文.
Bashir, M., Q. Bickle, H. Bushara, L. Cook, F. Shi, D. He, M. Huggins, J. Lin, K. Malik, A. Moloney, M. Mukhtar, P. Yeh, S. Xu, M. Taylor, and Y. Shi. 1994. Evalution of defined antigen vaccines against Schistosoma bovis and S. japonicum in bovines. Tropical and Geographical Medicine 46: 255-258.
Bergquist, N. R. 1995. Controlling Schistosomiasis by vaccination: A realistic option? Parasitology Today 11: 191-194.
Beverly L.M. and David A.D. 1986. Passive transfer with serum and IgG antibodies of irradiated cercaria-induced resistance against Schistosoma mansoni in mice. The Journal of Immunology. 136(7):2644-2648.
Beverly L.M., and David A.D. 1992. The role of IgG antibodies from irradiated cercaria-immunized rabbits in the passive transfer of immunity to Schistosoma mansoni-infected mice. Am. J. Trop. Med. Hyg. 47(6): 821-829.
Bickle, Q. D., B. J. Andrews, M. J. Doenhoff, M. J. Ford, and M. G. Taylor. 1985. Resistence against Schistosoma mansoni induced by highly irradiated infections:studies on species specificity of immunization and attempts to transfer resistance. Parasitology. 90:301-312.
Capron, A., J. P. Dessaint, M. Capron, J. H. Quma, and A. E. Butterworth. 1987. Immunity to Schistosomes:Progress to toward vaccine. Science 238: 1065-1072.
Capron, A., G. Riveau, J. M. Grzych, D. Boulanger, M. Capron, R. Pierce. 1994. Development of a Vaccine Strategy against human and bovine Schistosomiasis. Tropical and Geographical Medicane. 46 : 242-246.
Chen, M. G., and S. Fu. 1989. Schistosomiasis japonicum. In Tropical Medicine and Parasitology. Prentice-Hall International Inc. pp. 452-456.
Coles, G. C., J. I. Bruce, G. K. Kinoti, W. T. Mutahi, E. P. Dias, and N. Katz. 1986. Drug resistance in schistosomiasis. Transaction of the Royal Society of Tropical Medicine and Hygiene 80: 347.
Comoy, E. E.,C. Vendeville, A. Capron, and G. thyphronitis. 1997. Infection with Salmonella typhimurium Modulates the Immune Response to Schistosoma mansoni Glutathione-S-Transase. American Society for Microbiology. 65 : 3261-3266.
Comoy, E. E., A. Capron, and G.Thyphronitis. 1998. Adjuvant is the Major Parameter Influencing the Isotype Profiles Generated During Immunization with a Protein Antigen, the Schistosoma mansoni Sm 28-GST. Scand. J. Immunol. 47 : 444-452.
Doherty, P. J., M. H. Contreras, H. M. Dosch, and S. Pant. 1989. Rapid amplification of complementary DNA from small amounts of unfractionated RNA. Analytical Biochemistry 177: 7-10.
Dunne, D. W., F. M. Jones, L. Cock, and N. A. Moloney. 1994. Passively transferable protection against Schistosoma japonicum induced in the mouse by multiple vaccination with attenuated larvae: the development of immunity, antibody isotype responses and antigen recognition. Parasite Immunology 16: 655-668.
Duvall, R. H., and W. B. DeWitt. 1967. An improved perfusion technique for recovering adult schistosomes from laboratory animals. The American Journal of Tropical Medicine and Hygiene 16: 483-486.
Fallon, P. G., R. E. Fookes and M. J. Doenhoff. 1996. Protection of mice against Schistosoma mansoni infection by passive transfer of sera from infected rabbits. Parasite immunology 18:7-14.
Fan, P. C., K. M. Lee, C. C. Wu and C. Y. Liu. 1994. Further Study on Infectivity of Zoophilic and Anthropophilic Strains of Schistosoma Japonicum to Different Races of Oncomelania Hupensis Snails. Yonsei Reports on Tropical Medicine. 25 : 23-29.
Gobert, G. N., D. J. Stenzel, D. P. McManus. 1998. Immunolocalisation of the glutathione S-transferase, GST-26 and GST-28, within Schistosoma japonicum. International Journal for Parasitology 28 : 1437-1443.
Grzych, J. M., D. Grenzel, C. B. Xu, J. L. Neyrinck, M. Capron, J. H. Ouma, A. E. Butterworth, and A. Capron. 1993. IgA antibodies to a protective antigen in human Schistosomiasis mansoni . The Journal of immunology 150:527-535.
Henkle, K. J., K. M. Davern, M. D. Wright, A. J. Ramos, and G. F. Mitchell. 1990. Comparison of the cloned genes of the 26- and 28- kilodalton glutathione S-transferases of Schistosoma japonicum and Schistosoma mansoni. Molecular and Biochemical Parasitology 40: 23-34.
Hsu, H. F., S. Y. Li, C. K. Wang, P. C. Fan, and T. C. Huang. 1952. Studies on Schistosomiasis japonica in Formosa. American Journal of Tropical Medicine and Hygiene 1: 287-301.
Hsu, H. F., S. Y. Li Hsu, and L. S. Ritchie. 1955. Epidemiological study on Schistosomiasis japonica in Formosa. The American Journal of Tropical Medicine and Hygiene 4: 1042-1048.
Hsu, H. F., and S. Y. Li Hsu. 1956. On the infectivity of the Formosan strain of Schistosoma japonicum in Homo sapiens. American Journal of Tropical Medicine and Hygiene 5: 521-528.
Hsu, H. F., and S. Y. Li Hsu. 1962. Schistosoma japonicum in Formosa:A critical review. Experimental Parasitology 12: 459-462.
Janknecht, R., G. DeMartynoff, J. Lou, R. A. Hipskind, A. Nordheim, and H. G. Stunnenberg. 1991. Rapid and efficient purification of native histidine-tagged protein expressed by recombinant vaccine virus. Proceeding of the National Academy of Science USA 88: 8972-8976.
Lawson, B. W. L., Q. D. Bickle, and M. G. Taylor. 1995. Journal of Helminthology 69: 39-45.
Lee, K. M., C. T. Lo, P. C. Fan, and C. K. Chung. 1992. Susceptibility of Oncomelania hupensis subspecies to infection with Chinese, Taiwan and Japanese strains of Schistosoma japonicum. Chinese Journal of Parasitology 5: 67-71.
Liu, S. X., G. C. Song, Y. X. Xu, W. Yang, and D. P. Mc Manus. 1995.Anti-fecundity immunity induced in pigs vaccinated with recombinant Schistosoma japonicum 26 KDa glutathione-S-transferase. Parasite Immunology 17 :335-340.
Liu, S. X., G. C. Song, Y. X. Xu, Y. Wen, and D. P. Mc Manus. 1995. Immunization of mice with recombinant Sjc26GST induces a pronounced anti-fecundity effect after experimental infection with Chinese Schistosoma japonicum. Vaccine 13: 603-607.
Liu, S. X., Y. K. He, G. C. Song, X. S. Luo, Y. Xu, and D. P. Mc Manus. 1997. Anti-fecundity immunity to Schistosoma japonicum induced in Chinese water buffaloes(Bos buffelus)after vaccination with recombinant 26 KDa glutathione-S-transferase(reSjc26GST). Veterinary Parasitology 69 :39-47.
Liu, J. L., J. Fontaine, A. Capron and J. M. Grzych, 1996 Ultrastructural localization of Sm28 GST protective antigen in Schistosoma mansoni adulu worms . Parositology 113 : 377-391.
Lo, C. T., and C. Y. Lin. 1993. Infectivity of the newly-isolated Han-pao strain of Schistosoma japonicum from Taiwan to some oncomelanid snails. Annals of Tropical Medicine and Parasitology 87: 411-415.
Lo, C. T., and K. M. Lee. 1995. Schistosoma japonicum, zoophilic strain, in Oncomelania hupensis chiui and O. h. formosana: miracidial penetration and comparative histology. Journal of Parasitology 81: 708-713.
Mitchell, G. F., E. G. Garcia, K. M. Davern, W. U. Tiu, and D. B. Smith. 1988. Sensitization against the parasite antigen Sj26 is not sufficient for consistent expression of Schistosoma japonicum in mice. Transaction of the Royal Society of Tropical Medicine and Hygiene 82: 885-889.
Mitchell, G. F. 1989. Glutathione S-transferases-Potential components of anti-schistosome vaccines? Parasitology Today 5: 34-37.
Moloney, N. A., Q. D. Bickle, and G. Webbe. 1985. The induction of specific immunity against Schistosoma japonicum by exposure of mice to ultraviolet attenuated cercariae. Parasitology. 90 : 313-323.
Ohan, N. W., and J. J. Heikkila. 1993. Reverse transcription-polymerase chain reaction: An overview of the technique and its application. Advances in Biotechnology 11: 13-19.
O‘Leary, K. A., and J. W. Tracy. 1991. Schistosoma mansoni: Glutathione S-transferase-catalyzed detoxication of Dichlorvos. Experimental Parasitology 72: 355-361.
Owhashi, M., N. Kirai, Y. Hori. 1997. Eosinophii chemotactic factor release from neutrophils induced by stimulation with Schistosoma japonicum eggs. Parasitol Res. 83 : 42-46.
Pearce, E. J., S. L. James, S. Hiney, D. E. Lanar, A. Sher. 1988. Induction of protective immunity against Schistosoma manson by vaccination with schistosome paramyosin(Sm97), a nonsurface parasite antigen. Proc. Natl. Acad Sci. 85 : 5678-5682.
Price, H. P., M. J. Doenhoff, J. R. Sayers. 1996. Cloning, heterologyous expression and antigenicity of a schistosome cercarial proteases. Parasitology, 114 : 447-453.
Richter, D., R. N. Incani, and D. A. Harn. 1993. Isotype responses to candidate vaccine antigens in protective sera obtained from mice vaccinated with irradiated cercariae of Schistosoma mansoni. Infection and Immunity. 61 : 3003-3011.
Ross, A. G. P., Y. S. Li, A. C. Sleigh, and D. P. McManus. 1997. Schistosomiasis control in the people¢s republic of china. Parasitology Today 13:152-155.
Savioli, L., E. Renganathan, A. Montresor, A. Davis and K. Behbehani. 1997 Control of schistosomiasis-A Global Picture. Parasitology Today 13: 444-448.
Sher A., S. R. Smithers, and P. Mackenzie. 1975. Passive transfer of acquired resistance to Schistosoma mansoni in laboratory mice. Parasitology. 70 : 347-357.
Sher, A., S. R. Smithers, P. Mackenzie, and K. Broomfield. 1977. Schistosoma mansoni: Immunoglobulins involved in passive immunization of laboratory mice. Experimental Parasitology. 41 : 160-166.
Smith, D. B., K. M. Davern, P. G. Board, W. U. Tiu, E. G. Garcia, and G. F. Mitchell. 1986. Mr 26000 antigen of Schistosoma japonicum recognized by resistant WEHI 129/J mice is a parasite glutathione S-transferase. Proceeding of National Academy of Science USA. 83: 8703-8707.
Smith, M. A., J. A. Clegg, D. Snary, and A. J. Trejdosiewicz. 1982. Passive immunization of mice against Schistosoma mansoni with an IgM monolonal antibody. Parasitology. 84 : 83-91.
Smither, S. R. 1986. Improving prospects for a schistosomiasis vaccine. Nature 323: 205.
Taylor, M. G., M. C. Huggins, S. Fuhui, J. Lin, T. E, Y. Ping, S. Wei, Q. C. Gui, L. B. Fa, and Q. D. Bickle. 1998. Production and testing of Schistosoma japonicum candidate vaccine antigens in the natural ovine host. Vaccine. 16 : 1290-1298.
The QIAexpressionist: The high level expression and protein purification system. 1992. QIAGEN Inc.
Trottein, F., M. P. Kieny, C. Verwaerde, G. Torpier, R. J. Pierce, J. M. Balloul, D. Schmitt, J. P. Lecocq, and A. Capron. 1990. Molecular cloning and tissue distribution of a 26-kilodalton Schistosopma mansoni glutathione S-transferase. Molecular and Biochemical Parasitology 41: 35-44.
Trottein, F., C. Gordin, R. J. Pierce, B. Sellin, M. G. Taylor, I. Gorillot, M. S. Silva, J. P. Lecocq, and A. Capron. 1992. Inter-species variation of schistosome 28-KDa glutathione S-transferases. Molecular and Biochemical Parasitology 54: 63-72.
Victoratos, P., M. Yiagou, N. Avramidis, and L. Hadjipetrou. 1997. Regulation of cytokine gene expression by adjuvants in vivo. Clinical and Experimental Immunology. 109 : 569-578.
Walker, J., P. Crowley, A. D. Moreman, and J. Barrett. 1993. Biochemical properties of cloned glutathione S-transferase from Schistosoma mansoni and Schistosoma japonicum. Molecular and Biochemical Parasitology 61: 255-264.
Wang, J. S., F. M. Wu, K. C. Tung, and J. P. Chan. 1988. An outbreak and epidemiology of bovine schistosomiasis japonica in changhua county of Taiwan. Journal of Chinese Society of Veteranary Science 14: 298-296.
Wright, M. D., K. M. Davern, and G. F. Mitchell. 1991. The functional and immunological significance of some schistosome surface molecules. Parasitology Today 7: 56-58.
Wu, Z., Z. Shaoji, B. Pan, L. Hu, R. Wei, Z. Gao, J. Lie, and B. Uwe. 1994. Reinfection with Schistosoma japonicum after treatment with praziquantel in Poyang lake region, China. Southeast Asian Journal of Tropical Medicine and Public Health 25: 163-169.
Xu, C. B., C. Verwaerde, J. M. Grzych, J. Fontaine, and A. Capron. 1991. A monoclonal antibody blocking the Schistosoma mansoni 28KDa glutathione S-transferase activity reduces female worm fecundity and egg viability. Europe Journal of Immunology 21: 1801-1807.
Yan B.S., Lee K.M., Liu S.H., and Syu W.J. 1996. Characterization of monoclonal antibodies to the 26-KDa Glutathione S-transferase of Schistosoma japonicum. Hybridoma 15(6): 429-433.

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1. 黃世輝,1994,〈地方振興與現代設計〉,《臺灣手工業》,臺灣省手工業研究所,頁64-69。
2. 陳嬡蘋,1998,〈藝壇瑰寶,臺灣工藝的先驅者──顏水龍逝世專題報導〉,《文化視窗》,創刊號,頁36-41。
3. 陳樹升,1997,〈藝術探索──顏水龍的藝術世界〉,《明道文藝》,第258期,頁4-9。
4. 陳炎鋒,1985,〈顏水龍與花都半個世紀的戀情〉,《藝術家》,第127期,頁170-175。
5. 莊素娥,1992,〈純藝術的反叛者──顏水龍〉,《藝術家》,頁302-315。
6. 林俊成,1997,〈臺灣CIS的先驅──顏水龍〉,《臺灣美術》,頁5-8。
7. 李季育,1998,〈十九世紀末歐洲的「藝術與工藝運動」〉,《國立歷史博物館館刊(歷史文物)》,8:6 民87.06 頁32-36。
8. 吳婷玉,1996,〈老輩藝術家剪影──人老心不老:顏水龍〉,《典藏藝術雜誌》,第41期,頁134-137。
9. 江韶瑩,1992,〈臺灣工藝的發展與變遷(中)〉,《臺灣美術》,第15期,頁56-60。
10. 江韶瑩,1992,〈臺灣工藝的發展與變遷(下)〉,《臺灣美術》,第16期,頁85-90。
11. 江韶瑩,1991,〈臺灣工藝的發展與變遷(上)〉,《臺灣美術》,第14期,頁22-43。
12. 黃世輝,1998,〈談日本區域文化產業振興政策〉,《空間》,第107期,頁35-37。
13. 黃佩君,1993,〈讓藝術走出象牙塔──國寶級藝師顏水龍的藝術拓荒生涯〉,《中央月刊》,頁37-42。
14. 黃金財,1996,〈以藝載道的藝術人生──畫壇耆宿顏水龍〉,《臺灣月刊》,第165期,頁62-65。
15. 楊照,1997,〈捨棄傳統象牙塔,顏水龍回歸蔗民生活〉,《新新聞週刊》,第558期,頁80-81。
 
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