臺灣博碩士論文加值系統

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

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�@圖譜......................44
圖二、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49天之
糞便中第一期幼蟲蟲數變化情形...........................................45
圖三、以ELISA法檢查各組大鼠抗廣東住血線蟲雄性成蟲體抗原之血清抗體力價........46
圖四、以ELISA法檢查各組大鼠抗廣東住血線蟲雌性成蟲體抗原之血清抗體力價........47
圖五、以ELISA法檢查各組大鼠抗廣東住血線蟲雌性成蟲腸道蛋白質之血清抗體力價....48
圖六、免疫雄性成蟲體抗原之大鼠血清認知各抗原組成分子量位置...................49
圖七、免疫雌性成蟲體抗原之大鼠血清認知各抗原組成分子量位置...................50
圖八、免疫雌性成蟲腸道蛋白之大鼠血清認知各抗原組成分子量位置.................51
圖九、控制組之大鼠血清認知各抗原組成分子量位置...............................52
圖十、二級抗體認知各抗原組成分子量位置.......................................53
圖十一、以間接免疫螢光法分析免疫腸道蛋白質大鼠血清中抗體作用在廣東住血線蟲成蟲上
的位置.................................................................54

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