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標題: 胸膜肺炎放線桿菌RTX毒素之致病性
Pathogenesis of Actinobacillus pleuropneumoniae RTX-Toxins
作者: 張淑芬
Chang, Shu-Fen
關鍵字: Actinobacillus pleuropneumoniae
出版社: 獸醫學系
摘要: 胸膜肺炎放線桿菌引起豬放線桿菌胸膜炎,造成豬場嚴重之經濟損失。本 菌有許多分泌之物質、外膜性結構,及蛋白質毒力因子。其中, Actinobacillus pleuropneumoniae RTX-toxins (Apx)被認為是最重要的 。許多報告指出,Apx很可能是造成肺臟出血之主因。本研究即在探討Apx 對肺臟組織之傷害。以血清型第一型放線桿菌胸膜肺炎菌,利用膠體層析 過濾法來純化Apx,以蛋白質電泳分析以確定其分子量。以胸膜肺炎放線 桿菌、粗製及純化之Apx,分別對動物進行試驗。以間接免疫螢光法加以 觀察,並以組織病理學、穿透式電子顯微鏡觀察,以了解Apx是否即為造 成本病在肺臟出血的主要毒力因子。在臨床及實驗病例可見肺臟腫脹,觸 感堅硬,顏色暗紅,大小、形狀皆不規則之肉眼病變;組織切片下則可見 出血、多形態單核細胞浸潤於肺壞死區及肺泡腔內。穿透式電子顯微鏡觀 察時可見血管腔內聚集了許多紅血球,多數之血管細胞壞死的情形很嚴重 ,細胞核呈濃縮或已溶解,細胞質多數呈現空泡變化,並具有粒線體腫脹 、內質網擴張等變性變化。將該肺臟病變區製作冷凍切片,以間接免疫螢 光法加以觀察,結果病採對抗Apx-抗體呈陽性反應。結果顯示出,Apx確 實與本病在肺臟之出血具有相關性。
Actinobacillus pleuropneumoniae causes porcine Actinobacillus pleuropneumonia, a disease of great economic importance. Several secretory products and outer membrane components, of which Actinobacillus pleuropneumoniae RTX-toxins (Apx) were considered the most important. There were reported papers also indicated that Apx might be the major factor to induce hemorrhage. The purpose of this study was to gain further evidence of the Apx toxicity.The heat-sensitive Apx of Actinobacillus pleuropneumoniae serotype 1 strain 4074 was partially purified by ammonium sulfate precipitation and gel chromatography, then subjected to sodium dodecyl sulfatepoly-acrylamide gel and stained with Coomassie blue. In order to compare the lesions caused by rough or partially purified Apx, specific pathogen free pigs and guinea pigs were inoculated with Actinobacillus pleuropneumoniae . The clinical signs, gross lesions, histopathologic and ultrastructural changs were compared. Besides, by indirect immunofluorescent labelling the location of Apx in lung lesion was detected. The results showed that the lung lesions, induced by Actinobacillus pleuropneumoniae, rough Apx in pigs, or purified Apx in guinea pigs, were similar to those in clinical cases , included hemorrhage, necrosis, exudate, fibrin, and bacterial clumps in alveolar spaces . It could be detected the Apx localized in lung by indirect immunofluorescent immunostaining, both clinical and Apx inoculated cases presented positive reaction of anti-Apx antibody.The results supported that Apx could induce hemorrhage in lung.
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