Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/15153
標題: 假性狂犬病病毒感染對豬隻肺泡巨噬細胞、支氣管淋巴結及周邊血液單核細胞之影響
The Effect of Pseudorabies Virus Infection on Swine Alveolar Macrophages, Tracheobronchial Lymph Node, and Peripheral Blood Mononuclear Cells
作者: 劉俊旻
Liu, Chung-Min
關鍵字: pseudorabies
假性狂犬病
pulmonary alveolar macrophage
tracheobronchial lymph node
peripheral blood mononuclear cell
肺泡巨噬細胞
支氣管淋巴結
周邊血液單核細胞
出版社: 獸醫病理學研究所
摘要: 為了解感染假性狂犬病(Pseudorabies; PR)病毒對肉豬呼吸道微環境之免疫細胞與周邊相關免疫組織之影響,本實驗將PR病毒(TW strain; 2 ×106 TCID50)以氣管內接種感染豬隻,利用肺臟灌洗技術、免疫螢光染色及流式細胞儀以偵測巨噬細胞表面抗原的變化並分析其吞噬功能及活化狀態。在血液方面則以雙重染色分析PR病毒感染後周邊血液淋巴細胞的變化並進行中和抗體檢測,此外於各攻毒時間分批將實驗豬隻犧牲,進行病理學檢查及分析支氣管淋巴結內淋巴細胞次族群的變化。試驗結果顯示在PR病毒感染後第4天,PR病毒攻毒豬隻出現精神不振、食慾減退、口鼻有較多的分泌物及呼吸症狀等現象並逐漸嚴重,於攻毒後第11天PR感染豬隻逐漸恢復精神及食慾。攻毒豬隻在病理病變及組織病變上呈現非化膿性腦膜腦炎、肺臟有間質性肺炎及局部性多發壞死灶、支氣管淋巴結腫大及壞死灶。支氣管淋巴結於發病期間在切片下之T細胞依賴區呈現淋巴芽母細胞增生現象,而恢復期豬隻有明顯二次性淋巴濾泡增生。對照組及攻毒組豬隻於活體肺臟灌洗中的肺泡巨噬細胞於攻毒後第4及弟8天其吞噬 Pasteurella multocida的能力皆下降。但在感染後第11天,對照組豬隻肺泡巨噬細胞之吞噬能力雖有20.3﹪抑制但已接近恢復狀態,而PR攻毒組仍有50.4﹪的抑制現象。在肺泡巨噬細胞表現型上,對照及攻毒兩組之MHC class I及MHC class II分子於第8天都有揚升的情形。急性感染期間兩組之白血球細胞數雖有逐漸下降的情形,但兩組間曲線的變化極為類似。在抗體檢測方面,攻毒後第11之豬隻於血清及支氣管肺泡腔灌洗液的10倍濃縮液皆可測得中和抗體揚升,而對照組豬隻則仍低於2倍。攻毒組之周邊血液淋巴細胞CD4+CD8-次族群於PRV感染後第32天有顯著揚升,但兩處理組於各時間點CD4+及CD8+次族群都沒有顯現CD25分子有增加的情形。支氣管淋巴結淋巴細胞之IL-2R、MHC class II分子於感染後第4及第8天無明顯變化,但於第11天對照組及攻毒組支氣管淋巴結CD4+MHC class II+淋巴細胞次族群之相對比例揚升。感染後第32天攻毒組之淋巴結CD4+CD8+及CD4+MHC class II+淋巴細胞次族群之比例皆有明顯上升。以上結果顯示出PR病毒感染對於肺泡巨噬細胞之吞噬作用有明顯抑制作用,並且會延遲豬隻PAM恢復原本吞噬的能力,同時誘導支氣管淋巴結T細胞依賴區淋巴芽母細胞的活化,而在攻毒後第32天可見支氣管淋巴結有明顯的淋巴濾泡的增生及可檢測到攻毒豬隻其血清及支氣管肺泡腔灌洗液的10倍濃縮液的中和抗體明顯的揚升。但於實驗結果中支氣管淋巴結及周邊血液單核細胞淋巴細胞表現型變化的情形與支氣管淋巴結之組織病變下芽母細胞活化之觀察未能一致,其間相關的原因仍有待進一步之探討。
To investigate the effect of pseudorabies (PR) virus on pulmonary microenvironment and local immune responses, the experimental pigs (10-week-old) were endobronchially inoculated with PR virus (TW strain, 2 x 106 TCID50) or sterile PBS. At the designed time the bronchoalveolar lavages (BAL), blood samples, and regional lymph nodes were collected for phenotypic and phagocytic assays and pathological study. Pigs inoculated with PRV showed depression, anorexia, and slight respiratory distress on 4 days postinfection (dpi) and those symptoms were persistent until 11 dpi. Histopathological lesions on PRV infected pigs displayed mild nonsuppurative meningoencephaltis, broncho-interstitial pneumonia with mild focal necrosis at 4, 8, and 11 dpi. The trachobronchial lymph node showed focal necrosis and lymphoblast hyperplasia in T cell dependent areas on 8 and 11 dpi, and then hyperplasia of lymph follicles on 32 dpi. The neutralizing antibody in blood and concentrated BAL fluid were present on 7 and 11 dpi, respectively. Studies on phenotype of PAM showed an increase in the expression of MHC class I and MHC class II in both control and PRV infected pigs 4 dpi. The phagocytic function of PAM to P. multocida on both control and PRV-infected pigs was suppressed after endobronchial inoculation, but the effect was more serious and recovered slowly on PRV infected pigs than controls. When we looked at the changes and the activation of lymphocytic subpopulations of blood and middle tracheobronchial lymph node, a dual staining of lymphocytes was performed. The relative percentages of CD4 and CD8 subpopulation during acute PR virus infection were not significantly changed. However, the subpopulation of CD4+CD8+ were markedly increased at recovery stage of PRV infection. Moreover, the results also exhibited more CD4+ and CD8+ lymphocytes expressing MHC class II molecules after PRV infection in blood and regional lymph node. In contrast, the expression of IL-2R was not significantly increased in CD4+ and CD8+ lymphocytes during acute infection. Those data conclude that the suppressive effect of phagocytic function of PAM during acute PR virus infection may contribute to the secondary infection in pulmonary disease. The increase in CD4+CD8+ and more MHC class II expression on lymphocytes after PRV infection may reflect the activated state of lymphocytes during acute phase and then return to memory state at recovery stage. However, the less expression of activation marker, IL-2R, on lymphocytes during acute phase that was not matched to the pathological changes of node is remained further elucidated.
URI: http://hdl.handle.net/11455/15153
Appears in Collections:獸醫病理生物學所

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