Responsive image
博碩士論文 etd-0626101-125625 詳細資訊
Title page for etd-0626101-125625
論文名稱
Title
廣東住血線蟲成蟲腸道蛋白質之免疫力分析
Immunological analyses of intestinal proteins extracted from adult Angiostrongylus cantonensis
系所名稱
Department
畢業學年期
Year, semester
語文別
Language
學位類別
Degree
頁數
Number of pages
54
研究生
Author
指導教授
Advisor
召集委員
Convenor
口試委員
Advisory Committee
口試日期
Date of Exam
2001-06-18
繳交日期
Date of Submission
2001-06-26
關鍵字
Keywords
免疫保護力、腸道蛋白質、廣東住血線蟲
Angiostrongylus cantonensis, gut protein, protective immunity
統計
Statistics
本論文已被瀏覽 5699 次,被下載 8258
The thesis/dissertation has been browsed 5699 times, has been downloaded 8258 times.
中文摘要
本研究以廣東住血線蟲雄性成蟲體抗原、雌性成蟲體抗原及雌性成蟲腸道蛋白質免疫SD品系大鼠,評估腸道蛋白質在作為保護免疫原上的價值。結果在腦部第五期幼蟲及心肺部成蟲回收方面,其保護率分別為15 %及14 %,較其他各免疫組來得低,但在回收的蟲體長度方面,腸道蛋白質組蟲體長度則較其他各組來得短,另外在大鼠每克糞便中第一期幼蟲的排出上,腸道蛋白質組亦較低。在細胞性免疫反應的偵測上,以腸道蛋白質刺激大鼠脾臟淋巴母細胞時,呈現隨免疫次數增加刺激指數也增加的情形,且其刺激指數較其他各組來得高,在血清中IgG抗體的效價和免疫保護力沒有明顯的相關性。各抗原組成中一分子量為84kDa之蛋白質分子對腸道蛋白質的免疫血清均出現強烈的抗原辨識性。以間接免疫螢光法偵測,發現在成蟲的肌肉層及腸道和生殖道的外層以及腸道內腔出現顯著的螢光反應。另外腸道蛋白質和腸道膜蛋白的組成相當類似,各組免疫血清認知腸道蛋白質組成的分子量位置大部分亦出現在腸道膜蛋白的組成上,因此腸道膜蛋白及84kDa蛋白質在對廣東住血線蟲感染的保護免疫力上所扮演的角色,需要更進一步的研究。
Abstract
Abstract

In order to determine whether antigens prepared from adult intestine of Angiostrongylus cantonensis have potential to induce a protective immunity in the rodent hosts, somatic antigens extracted from male and female adults as well as gut antigens isolated from female adults were used to immunize rats against A. cantonensis infection. A 14% reduction in L5 recorvery from brain as well as a 15% reduction in adult recovery from pulmonary artery were achieved in the immunized rats when compared with the control group after infection with 50 larvae. The length of worms recovered from immunized rats was shorter than that in other groups. The larvae recovered from fecal materials in immunized rats were also reduced. In cell proliferation test, the stimulation index of gut antigens increased with times of immunization and exhibited the highest values. However, serum IgG titers were not correlated with protective immune responses. A 84 kDa protein contained in all antigen preparations was recognized by immune serum against gut antigens. Strong positive reactions were detected by indirect immunofluorescent assay in the internal musculature of the body, gut and reproductive tract wall and gut lumen. The composition of gut antigens was similar to that of gut membrane proteins. Immune sera recognized several major gut proteins were also appeared on gut membrane proteins. Further studies are required to provide evidence that gut membrane proteins play in the protective immune response against A. cantonensis infections, including the 84 kDa protein.
目次 Table of Contents
目錄

前言…………………………………………………………………1
材料與方法…………………………………………………………5
結果………………………………………………………………..16
討論………………………………………………………………..25
參考文獻…………………………………………………………..30
附表………………………………………………………………..36
附圖………………………………………………………………..44

圖表目錄

表一、以廣東住血線蟲雄性成蟲體抗原、雌性成蟲體抗原及雌性成蟲腸道蛋白免疫後,再以
50條第三期幼蟲感染之大鼠,其腦部第五期幼蟲之回收情形...................36
表二、以廣東住血線蟲雄性成蟲體抗原、雌性成蟲體抗原及雌性成蟲腸道蛋白免疫後,再以
50條第三期幼蟲感染之大鼠,其心肺部成蟲之回收情形.......................37
表三、以廣東住血線蟲雄性成蟲體抗原、雌性成蟲體抗原及雌性成蟲腸道蛋白免疫後,再以
50條第三期幼蟲感染之大鼠,其腦部第五期幼蟲之蟲體長度變化情形...........38
表四、以廣東住血線蟲雄性成蟲體抗原、雌性成蟲體抗原及雌性成蟲腸道蛋白免疫後,再以
50條第三期幼蟲感染之大鼠,其心肺部成蟲之蟲體長度變化情形...............39
表五、各組大鼠於二次免疫後,感染50隻廣東住血線蟲第三期幼蟲,其感染後35天到50天之
糞便中第一期幼蟲蟲數...................................................40
表六、廣東住血線蟲雄性成蟲體抗原刺激免疫後大鼠脾臟淋巴母細胞增殖反應結果.....41
表七、廣東住血線蟲雌性成蟲體抗原刺激免疫後大鼠脾臟淋巴母細胞增殖反應結果.....42
表八、廣東住血線蟲雌性成蟲腸道蛋白刺激免疫後大鼠脾臟淋巴母細胞增殖反應結果...43
圖一、廣東住血線蟲四種不同抗原經SDS膠體電泳後之胜
參考文獻 References

Alger N. E., Cabrera E. J. (1972) An increase in death rate of Anopheles stephensi fed on rabbits immunized with mosquito antigen. Journal of Economic Entomology 65: 165-168
Alicata J. E. (1988) Angiostrongyliasis cantonensis (eosinophilic meningitis): historical events in its recognition as a new parasitic disease of man. Journal of Washington Academy Science 78: 38-46
Alicata J. E., Brown R. W. (1962) Observations on method of human infection with Angiostrongylus cantonensis in Tahiti. Canadian Journal of Zoology 40: 755-760
Alicata J. E., Jindrak K. (1970) Angiostrongylosis in the Pacific and South-East Asia. Charles C. Thomas, Springfield, Illinois. pp. 105
Bhaibulaya M. (1975) Comparative studies on the life cycle of angiostrongylus mackerrasae Bhaibulaya, 1968 and Angiostrongylus cantonensis Chen, 1935. International Journal for Parasitology 5: 7-20
Bitakaramire P. K. (1966) Studies on immunity to Haemonchus contortus infection: the elimination of a challenge infection by immune sheep. Parasitology 56: 619-622
Chen S. N. (1986) Enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies to Angiostrongylus cantonensis. Transactions of the Royal Society of Tropical Medicine and Hygiene 80: 398-405
Cheng C. W., Sato Y., Chen E. R. (1984) Epidemiological and clinical observations on eosinophilic meningitis and meningoencephalitis of children caused by infection with Angiostrongylus cantonensis in southern Taiwan. Ryukgu Medical Journal 7: 1-9
Dharmkrong-at A., Sirisinha S. (1983) Analysis of antigens from different developmental stages of Angiostrongylus cantonensis. The Southeast Asian Journal of Tropical Medicine and Public Health 14: 154-162
Fujii T. (1987) Angiostrongylus cantonensis: immunoblot analysis of the antigens recognized by rats. Parasitology Research 73: 366-374
Jasmer D. P., McGuire T. C. (1991) Protective immunity to a blood-feeding nematode (Haemonchus contortus) induced by parasite gut antigens.
Infection and Immunity 59: 4412-4417
Kamiya M. (1970) Haemagglutination test in rats infected with Angiostrongylus cantonensis. Southeast Asian Journal of Tropical Medicine and Public Health 1: 570-571
Kamiya M., Klongkamnuankarn K., Tharavanij S., Tanaka H. (1972) Change of indirect hemagglutination reactions in serum after the transfer of adult Angiostrongylus cantonensis to the abdominal cavity of rats. The Southeast Asian Journal of Tropical Medicine and Public Health 3: 119-123
Kamiya M., Tanaka H. (1969) Hemagglutination test in rats infected with Angiostrongylus cantonensis. Japanese Journal of Experimental Medicine 39: 593-599
Knox D. P., Smith S. K., Smith W. D.(1999) Immunization with an affinity purified protein extracted from the adult parasite protects lambs against infection with Haemonchus contortus. Parasite Immunology 21: 201-210
Ko R. C., Chiu M. C., Kum W., Chan S. H. (1984) First report of human angiostrongyliasis in Hong Kong diagnosed by computerized axial topography (CAT) and enzyme-linked immunosorbent assay. Transection of the Royal Society of Tropical Medicine and Hygiene 18: 354-355
Kum W. W., Ko R. C. (1987) Active and passive immunity against Angiostrongylus cantonensis infection in mice and rats. Tropical Biomedicine 4: 73-74
Lee S. H. (1969) The use of irradiated third-stage larvae of Angiostrongylus cantonensis as antigen to immunize albino rats against homologous infection. Proceedings of Helminthology Society of Washington 36: 95-97
Lee J. D., Chen E. R., Chung L. Y., Yen C. M. (1997) Humoral immune responses in mice infected with gamma-irradiated third-stage larvae of Angiostrongylus cantonensis. Kaohsiung Journal of Medical Science 13: 277-85
Liang H. K., Jin X. X., Feng Z. M., Wang M. Y., Pan S. T., Su S. Z.(1993) Detection of serum antibodies in rats infected with Angiostrongylus cantonensis by indirect fluorescent antibody test. Chinese Journal of Parasitology and Parasitic Diseases 11: 286-287
Lim B. L., Kong O. C., Joe L. K.(1965) Natural infection of Angiostrongylus cantonensis in Malaysian rodents and intermediated hosts, and preliminary observations on acquired resistance. American Journal of Tropical Medicine and Hygiene 14: 610-617
Lok J. B., Knight D. H., Selavka C. M., Eynard J., Zhang Y., Bergman R. N. (1995)Studies of reproductive competence in male Dirofilaria immitis treated with milbemycin oxime. Tropical Medicine and Parasitology 46:235-40
Munn E. A., Smith T. S., Smith H. et al. (1997) Vaccination against Haemonchus contortus with denatured forms of the protective antigen H11. Parasite Immunology 19: 243-248
Nomura S., Lin P. H. (1945) First case of human infection with Haemostrongylus ratti Yokogawa. Taiwan No Ikai 3: 589-592
Opdebeeck J. P., Wong J. Y., Jackson L. A., Dobson C. (1988) Hereford cattle immunized and protected against Boophilus microplus with soluble and membrane-associated antigens from the midgut of ticks. Parasite Immunology 10: 405-10
Prieto G., McCall J. W., Venco L., Genchi M., Simon F., Genchi C. (2001) IgG response against infective larvae of Dirofilaria immitis in experimentally infected cats. Veterinary Research 32: 93-96
Shaio M. F., Hou S. C., Chen J. G. (1990) Immunoglobulin G-dependent classical complement pathway activation in neutrophil-mediated cytotoxicity to infective larvae of Angiostrongylus cantonensis. Annals of Tropical Medicine and parsitology 84: 185-191
Smith W. D. (1993) Protection in lambs immunised with Haemonchus contortus gut membrane proteins. Research in Veterinary Science 54: 1-9
Smith S. K., Pettit D., Newlands G. F., Redmond D. L., Skuce P. J., Knox D. P, Smith W. D. (1999) Further immunization and biochemical studies with a protective antigen complex from the microvillar membrane of the intestine of Haemonchus contortus. Parasite Immunology 21: 187-99
Smith W. D., Smith S. K., Murray J. M. (1994) Protection studies with intergral membrane protein fractions of Haemonchus contortus. Parasite Immunology 16: 231-241
Sugaya H., Abe T., Yoshimura K., Sasaki O. (1993) Antigen dependent release of interleukin 5 in vitro from spleen cells of mice infected with Angiostrongylus cantonensis. International Journal for Parasitology 23: 865-9
Sugaya H., Ishida K., Yoshimura K. (1997) Induction of cerebrospinal fluid eosinophilia in rats by the intraventricular injection of Angiostrongylus cantonensis antigen. International Journal for Parasitology 27: 113-117
Techasoponmani R., Sirisinha S. (1980) Use of excretory and secretory products from adult female worms to immunize rats and mice against Angiostrongylus cantonensis infection. Parasitology 80: 457-469
Uahkowithchai V., Techasoponmani R., Sirisinha S. (1977) Effect of immunization with metabolic antigens on experimental infection of rats with Angiostrongylus cantonensis. Southeast Asian Journal of Tropical Medicine and Public Health 8: 486-493
Wang L. C., Chao D., Chen E. R. (1989) Acquired immunity in rats against Angiostrongylus cantonensis infections. International Journal for Parasitology 19: 617-620
Welch J. S., Dobson C. (1978) Immunodiagnosis of parasitic zoonoses: comparative efficacy of three immunofluorescence tests using antigens purified by affinity chromatography. Transactions of the Royal Society of Tropical Medicine and Hygiene 72: 282-288
Willadsen P., Riding G. A., McKenna R. V., Kemp D. H., Tellam R. L., Nielsen J. N., Lahnstein J., Cobon G. S., Gough J. M.(1989) Imuunological control of a parasitic arthropod. Identification of a protective antigen from Boophilus microplus. The Journal of Immunology 143:1346-51
Yii C. Y. (1976) Clinical observations on eosinophilic meningitis and meningoencephalitis caused by Angiostrongylus cantonensis on Taiwan. American Journal of Tropical Medicine and Hygiene 25: 233-249
Yong W. K., Dobson C. (1982a) Population dynamics of Angiostrongylus cantonensis during primary infections in rats. Parasitology 85: 399-409
Yong W. K., Dobson C. (1982b) Antibody responses in rats infected with Angiostrongylus cantonensis and the passive transfer of protective immunity with immune serum. Zeitschrift fur Parasitenkunde 67: 329-336
Yong W. K., Glanville R. J., Dobson C. (1983) The role of the spleen in protective immunity against Angiostrongylus cantonensis in rats: splenectomy and passive spleen cell transfers. International Journal for Parasitology 13: 165-170
Yoshimura K., Aiba H., Oya H. (1979) Transplantation of young adult Angiostrongylus cantonensis into the rat pulmonary vessels and its application to the assessment of acquired resistance. International Journal for Parasitology 9: 97-103
Yoshimura K., Sato K., Uchida K., Oya H. (1982) The course of Angiostrongylus cantonensis infection in athymic nude and neonatally thymectomized mice. Zeitschrift fur Parasitenkunde 67: 217-226
Yoshimura K., Uchida K., Sato K., Oya H.(1983) Antibody-dependent adherence and damage to the young adult worms of Angiostrongylus cantonensis by rat eosinophils in vitro. Japanese Journal of Parasitology 32: 489-500
電子全文 Fulltext
本電子全文僅授權使用者為學術研究之目的,進行個人非營利性質之檢索、閱讀、列印。請遵守中華民國著作權法之相關規定,切勿任意重製、散佈、改作、轉貼、播送,以免觸法。
論文使用權限 Thesis access permission:校內外都一年後公開 withheld
開放時間 Available:
校內 Campus: 已公開 available
校外 Off-campus: 已公開 available


紙本論文 Printed copies
紙本論文的公開資訊在102學年度以後相對較為完整。如果需要查詢101學年度以前的紙本論文公開資訊,請聯繫圖資處紙本論文服務櫃台。如有不便之處敬請見諒。
開放時間 available 已公開 available

QR Code