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標題: 家禽里奧病毒足底接種後之足底病理變化及炎症細胞的消長
Pathological Changes of Footpad and Fluctuation of Inflammatory Cells after Arthrotropic Avian Reovirus Footpad Inoculation
作者: Tsung-Ching Liu
關鍵字: 家禽里奧病毒;S1133株;炎症反應;異嗜球;Avian reovirus;S1133 strain;inflammation;heterophil
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以往家禽里奧病毒所造成的病毒性關節炎在野外感染後所觀察的病變皆以跗關節為主。而學者們在實驗模式接種下(含足底)也依野外感染的方式觀察跗關節,卻少有人觀察足底接種後的足底病變。本實驗先將家禽里奧病毒S1133株進行病毒培養複製再計算力價後以2000 TCID50的劑量共10 μl接種左肢足底,並觀察足底的病理變化以及炎症細胞的消長情形,最後再以免疫組織化學染色去標定家禽里奧病毒的複製訊號以及FITC染色去標定異嗜球;以接種10 μl滅菌PBS當作控制組。結果可觀察到足底接種病毒後有潮紅腫脹的現象,但潮紅會在接種後第5至6天逐漸消退而只剩下腫脹。鏡檢下,充鬱血情形與肉眼病變結果相呼應。此外除了看到淺層滑膜及纖維滑膜細胞的活化後增生肥大外,也可在接種處及趾骨與趾關節交接處有炎症細胞浸潤,一開始以淋巴球浸潤為主,但接種後第2天則逐漸改以異嗜球浸潤,直到接種後第5天,轉呈現少量至中等量的淋巴球浸潤,此外另可在接種後第3天開始有淋巴濾泡於血管周圍形成,而接種處引起的炎症反應會比骨腱交接處嚴重,其原因可能與病毒量及血管分布的多寡有關。在使用FITC染色及免疫組織化學染色計算出異嗜球的絕對數量並進行病變指數評分,可見接種後1.5至3.5天異嗜球的數量達到最高而在接種後第6天開始只可見少數異嗜球;此外,異嗜球並非家禽里奧病毒複製場所,而足底的滑膜細胞才是其標的細胞。關節囊液會在接種後第1至6天觀察到大量異嗜球伴隨少量脫落的滑膜細胞,之後異嗜球大量消退,該現象與野外感染的跗關節內關節囊液所觀察到的結果相類似。為了釐清炎症反應及異嗜球的趨化非受到病毒培養之細胞質內容物及病毒顆粒本身所造成,另外進行左肢足底接種培養過細胞的培養液及使用紫外光不活化後的病毒液各10 μl,結果顯現除了在接種後第1至2天有觀察到輕微的局部炎症反應外,並無觀察到類似接種家禽里奧病毒等之滑膜細胞活化或異嗜球趨化等病變。故認為家禽里奧病毒接種後所引起的炎症反應是細胞受到病毒感染後所引起,且炎症細胞及滑膜細胞並不會特別受到細胞質內容物或是病毒顆粒本身而引致趨化及活化。然而要了解異嗜球在整個致病機轉上所扮演的角色,則有待後續研究進行。

In the past, the lesion observed in chicken infected by wild strand virus of avian reovirus was confirned to the tarsal joint. In experiment models, scholars observed tarsal joint leion rather than foot pad lesion even after footpad inoculation. The aim of this experiment is to understand the pathological changes of footpad and the fluctuation of inflammatory cells after arthrotropic ARV S1133 footpad inoculation. The footpad appeared red and swollen after day post-inoculation (DPI) 5-6 then only swelling was noticed. Hyperemia and congestion were evident microscopically wihich concur with redness of the gross lesion. In addition to synovial cells activation evident by hyperplasia and hypertrophy, there were inflammatory cells at the inoculation site and the area between digital bone and digital flexor tendon. At first, majority population of the cells were lymphocytes, then shifted to heterophils infiltration after DPI 2-5 and at last replaced with mild to moderate lymphocytic infiltration starting after DPI 6. Moreover, perivascular lymphoid nodules were formed after DPI 3. The lesion at the inoculation site was more severe than the area between digital bone and digital flexor tendon which might be related to virus quantity and vascular richness. The result of the FITC stain and immunohistochemical stain showed highest count of heterophils after DPI 1.5-3.5 and only few heterophils were seen after DPI 6. Furthermore, synovial cells were the target cells of ARV replication rather than heterophils. The synovial fluid consisted of heterophils and few exfoliated synovial cells after DPI 1-6 then followed by drastic decrease in count. Another experiment using cell medium (used for viral incubation) and inactivated viral fluid for inoculation was pursued in order to clarify whether the inflammation and heterophil chemotaxis were induced by cytoplasmic content/viral particles or viral infection. The result showed that there were no similar pathological changes except mild inflammation after DPI 1-2. Therefore, inflammation after inoculation was induced by ARV infection rather than cytoplasmic content or virus particles. However, the exact role of heterophils responsible for the pathogenesis of ARV infection needs further investigation.
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