Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/66266
標題: 天竺鼠肺泡巨噬細胞經假性狂犬病疫苗激發後之形態變化
Morphologic Changes of Stimulated Alveolar Macrophages in Guinea Pig due to Pseudorabies Vaccination
作者: 洪成均
Hung, Terence
關鍵字: Alveolar macrophage
肺泡巨噬細胞
Pseudorabies vaccine
Lysosome
Actinobacillus pleuropneumoniae
Phagocytosis
Intracellular degradation
假性狂犬病疫苗
溶素體
胸膜肺炎放線桿菌
吞噬作用
胞內降解作用
出版社: 獸醫微生物學研究所
摘要: 摘要 肺臟中之肺泡巨噬細胞( Alveolar macrophage)駐在肺泡表面 ,為肺臟防禦異物侵入之第一線防禦細胞,具有吞噬、消化細菌的能力, 也是肺泡腔能維持無菌狀態最主要的原因,為瞭解肺泡巨噬細胞對疫苗免 疫作用的反應,以增強肺泡巨噬細胞清除胸膜肺炎放線桿菌的能力,故本 實驗以油劑假性狂犬病疫苗,免疫天竺鼠,取其肺泡巨噬細胞,進行活體 外試驗,檢測其被激活的程度,細胞數目,細胞內溶素體的數量及吞噬能 力,比較受激活的和未受激活細胞之間的差異,並以形態學及定量的方法 ,利用光學及穿透式電子顯微鏡,直接觀察肺泡巨噬細胞受激活後之形態 變化,及吞噬消化胸膜肺炎放線桿菌的能力,以瞭解假性狂犬病疫苗免疫 對肺泡巨噬細胞的激活作用。 實驗結果顯示,受免疫的天竺鼠肺臟可 見其支氣管周圍之淋巴濾泡明顯增大 , 於對照組的淋巴濾泡平均直徑大 小為 71.66 ± 22.89 μm,而實驗組為194.13 ± 71.42 μm,兩組間具 有顯著差異(p<0.01);所沖出之肺泡巨噬細胞數目,以每克肺所獲得之 總平均細胞數來表示,對照組為2.14 ± 0.87 個,而實驗組為 5.95 ± 3.38個,免疫組較對照組顯著增多(P< 0.05),但是其增加的來源和血 液中單核球並沒有直接關係;計算溶素體與細胞質的面積比方面,對照組 為 0.021 ± 0.007,實驗組為 0.110 ± 0.032, 兩組間具有顯著差異 (P< 0.05),可見免疫後肺泡巨噬細胞內之溶素體數目增多;而加入胸 膜肺炎放線桿菌後,經穿透式電子顯微鏡的觀察,結果清楚的發現免疫組 肺泡巨噬細胞較未免疫的肺泡巨噬細胞可發揮較大的吞噬作用及較強的胞 內降解能力。 由這些形態學上的証據,客觀的證明經油劑假性狂犬病 疫苗免疫後的肺臟,在免疫防禦上有較佳的抵抗性;而受疫苗免疫激活的 肺泡巨噬細胞,對胸膜肺炎放線桿菌則有較強的清除力 。 由此推論經假 性狂犬病疫苗免疫後,肺泡巨噬細胞有受激活的效果,可使細胞本身之吞 噬消化功能加強,進而增加其對抗細菌的能力,達到預防胸膜肺炎放線桿 菌感染的目的。
Abstract The pulmonary alveolar macrophages reside on the alveolar surface , where they provide phagocytic defense against invading bacteria in the respiratory tract. They play a major role in the first defense and maintain the sterility of the alveolar spaces of lung. In order to understand the effects of vaccination on alveolar macrophages, guinea pigs were immunized with commercial oil type pseudorabies vaccine. Fourteen days later, the alveolar macrophages were collected by lung lavage and cultured in RPMI-1640 medium. The cells were counted in a hemocytometer and the number of intracytoplasmic lysosomes was measured by morphometric technique. Bacterial cultivation of Actinobacillus pleuropneumoniae was performed to assess the phagocytotic and the bacteriocidal ability at various time intervals, examed by light microscopy and electron microscopy. The purpose of this study was to observe the morphologic changes and phagocytotic activity of alveolar macrophages after the stimulation of pseudorabies vaccination. The results showed that the average diameter of bronchus- associated lymphoid follicles was 71.66 ± 22.89 μm in control group, and 194.13 ± 71.42 μm in vaccinated group (p<0.01). The average number of alveolar macrophages in the lung lavage fluid was 2.14 ± 0.78 X 106 cells/gm in control group,and 5.95 ± 3.38 X 106 cells/gm in vaccinated group(p<0.05 ). The mean area ratio of lysosomes over cytoplasm was 0.021 ± 0.007 in control group, and 0.110 ±0.032 in vaccinated group(p<0.05 ). The macrophage lysosomes of vaccinated group was more than that in control group. After challenging with Actinobacillus pleuropneumoniae, macrophage phagosomes in vaccinated group were more than that in control group. The phagocytic index of vaccinated group was significantly higher than control at 0.5,1, or 4 hours after inoculation. In the culture of macrophages with Actinobacillus pleuropneumoniae, degenerative changes appeared during first hour in control group, but occured at secondary hours and later in vaccinated group. Conclusively, the alveolar macrophages could be activated to enhance their microbiocidal ability by pseudorabies vaccination. These morphologic and functional changes were useful to clean the Actinobacillus pleuropneumoniae in lung. Therefore, it was considered that the vaccinated animals may obtain much ability to prevent from the Actinobacillus pleuropneumoniae infection.
URI: http://hdl.handle.net/11455/66266
Appears in Collections:微生物暨公共衛生學研究所

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