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系統識別號 U0026-0812200912094721
論文名稱(中文) 克沙奇B3病毒引起肝炎之體外及體內模式的研究
論文名稱(英文) In vitro and in vivo studies of coxsackievirus B3-induced hepatitis
校院名稱 成功大學
系所名稱(中) 微生物及免疫學研究所
系所名稱(英) Department of Microbiology & Immunology
學年度 94
學期 2
出版年 95
研究生(中文) 劉蓉燕
研究生(英文) Jung-Yen Liu
學號 s4693405
學位類別 碩士
語文別 英文
論文頁數 45頁
口試委員 召集委員-蘇益仁
口試委員-黎煥耀
指導教授-劉清泉
口試委員-余俊強
口試委員-林貴香
中文關鍵字 延遲加速因子  克沙奇病毒和腺病毒接受器  動物模式  肝炎  克沙奇B型病毒 
英文關鍵字 hepatitis  Coxsackievirus B  decay-accelerating factor  animal model  coxsackievirus and adenovirus receptor 
學科別分類
中文摘要 克沙奇B群病毒屬於人類腸病毒的一種,可在嬰幼兒的感染造成多樣的臨床症狀,包括輕微無特異性的發燒、皮疹、腸胃道症狀,到嚴重的肝炎、心肌炎和腦膜腦炎等、甚至會併發休克和瀰漫性血管內凝集,進而威脅生命導致多重器官衰竭死亡。在過去的研究報告顯示,致死性克沙奇B群病毒感染主要是引發心肌炎或腦膜腦炎。在台灣,近年來都有克沙奇B群病毒造成嬰幼兒死亡的案例,但在臨床上觀察發現,似敗血性症候群(sepsis-like syndrome)及猛爆性肝炎為致死的主要死因,且主要流行的病毒株為克沙奇B群病毒第三型。患者多為小於三個月的小嬰兒或新生兒,以發燒、黃疸、皮膚瘀血為表徵,實驗室數據可發現血小板低下、貧血、凝血時間延長、轉氨酶增加、膽紅素升高、及乳酸去氫酶(lactate dehydrogenase, LDH)大幅度升高等,除此之外克沙奇病毒B群病毒接受器延遲加速因子(decay accelerating factor)在克沙奇B型病毒病人的周邊血液單核球細胞的表現增加。因此本研究的目的是要探討克沙奇B群病毒侵犯肝細胞的趨向性,以及探討克沙奇病毒B群病毒接受器在致病機轉所扮演的角色。在體外實驗部份發現,從臨床分離出的克沙奇B群病毒可以在肝細胞生長,造成肝細胞的死亡,而且克沙奇B群病毒第三型較其他腸病毒比起來,傾向於侵犯肝臟,造成肝細胞的死亡。在動物實驗方面,七天大的小鼠以腹腔注射克沙奇B群病毒第三型兩小時後,即能在肝臟偵測到病毒的存在,之後病毒會散佈到其他許多器官,但仍以肝臟的病毒量最高。除此之外,小鼠感染克沙奇B群病毒後,也可以產生轉氨酶及乳酸去氫酶增加等類似的臨床症狀及病理變化。我們發現小鼠的肝臟會大量表現克沙奇病毒-腺病毒接受器(coxsackievirus-adenovirus receptor),且隨著年齡的升高有降低的趨勢,而且在體外模式中加入抗克沙奇病毒-腺病毒接受器的單株抗體可以阻斷克沙奇B型病毒去感染肝細胞。由以上結果顯示,克沙奇B型病毒有侵犯肝細胞的趨向性,以及克沙奇病毒B群病毒接受器在致病機轉中也扮演了一個重要的角色。這也讓我們更了解克沙奇B型病毒的致病過程與機制。

英文摘要 Coxsackievirus B (CB) belongs to the Picornaviridae family. It is responsible for an array of clinical diseases ranging from asymptomatic illness to severe diseases such as myocarditis, meningoencephalitis and fulminant hepatitis. In 2005 an outbreak of CB infection, which was characterized by fulminant hepatitis in neonates occurred in Taiwan. Patients with CB-associated fulminant hepatitis usually have abnormal liver function with markedly elevated aspartate aminotransferase (AST), alanine aminotransferase (ALT) and lactate dehydrogenase (LDH). Using clinical isolated strains, we found that CB3 grew in hepatocytes and induced cell death more severe than other enteroviruses. These observations also could be demonstrated in seven-day-old ICR mice. After intraperitioneal inoculation, CB3 was first detected in the liver. At the 48 hours post infection, virus could be isolated from many tissues, including brain, heart, lung, liver and spleen, with liver being the organ containing the highest viral titer. Histopathological studies illustrated neutrophilic infiltration and cell death in the liver. Furthermore, we found that decay accelerating factor (DAF), a CB receptor, was up-regulated on PBMC of CB patients. To exploit the possible mechanisms of CB3 entry, we examined the expression of the viral receptors in various mice organs. We found that coxsackievirus-adenovirus receptor (CAR) was highly expressed in the liver of newborn mice, and the expression was inversely correlated with age of mice. Anti-CAR antibody inhibited CB3 infection of liver cells in vitro, suggesting that the receptor plays an important role for mediating CB infection of hepatocytes. In summary, these findings provide further insight into the mechanism of CB3 to infect liver.

論文目次 Abstract i
Abstract in Chinese ii
Acknowledgment iiii
Contents v
Index of Figures vi
Abbreviations vii
Introduction 1
Materials and Methods 9
Results 16
Discussion 21
References 25
Appendix 1 44
Appendix 2 45
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