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    Title: SARS冠狀病毒類木瓜酵素對類鐸受體7及8訊號傳遞之影響
    Effect of SARS coronavirus papain-like protease on Toll-like receptor 7/8- signaling
    Authors: 蕭莉馨;Li-Hsin Hsiao
    Contributors: 健康照護學院醫學檢驗生物技術學系碩士班
    Keywords: 嚴重急性呼吸道症候群;類鐸受體7.8;SARS;Toll-like receptor 7/8
    Date: 2010
    Issue Date: 2010-09-29 12:10:21 (UTC+8)
    Abstract: 嚴重急性呼吸道症候群冠狀病毒為有高度傳染性,具套膜的正向單股RNA病毒,會引起致死性呼吸道疾病。SARS-CoV的基因組中的兩個大型多聚蛋白,分別為pp1a及pp1b,可編碼出兩個蛋白酶,分別為類木瓜酵素及3C-like蛋白酶。而類木瓜酵素被發現可抑制先天性免疫反應。在本篇研究,我們將利用類鐸受體7及8配位體咪喹莫特測試類木瓜酵素對類鐸受體7及8訊號傳遞的影響。首先,將類木瓜酵素構築在人類單核球白血病細胞中。再經實驗得知類木瓜酵素會抑制ISRE及NF-κB啟動子的活化。隨後利用即時定量聚合酶連鎖反應試驗可知類木瓜酵素會使IL-6,IL-8,PKR及OAS基因表現量下降。重要的是我們發現類木瓜酵素會影響訊號傳遞活化,如TRAF3及TRAF6泛素化能力下降,p38 MAPK,NF-κB,CREB,IRF-3, STAT1及JAK-2磷酸化受到不同程度的影響。在這篇論文結果中,我們可以了解嚴重急性呼吸道症候群冠狀病毒在宿主細胞的類鐸受體7及8訊號傳遞對所產生的影響。

    Severe acute respiratory syndrome coronavirus (SARS-CoV), an enveloped single-strand positive-sense RNA virus, is highly contagious, causing mortal respiratory disease. The SARS-CoV RNA genome encodes two different proteases papain-like protease (PLpro) and 3C-like protease (3CLpro), being responsible for proteolytic processes of large replicase polyproteins pp1a and pp1b. PLpro is reported to be an interferon antagonist, being associated with the inhibition of innate immune responses. In this study, we investigated the effects of PLpro on TRL7/8 signaling induced by imiquimod (IMQ). Firstly, expression of PLpro in human promonocyte HL-CZ cells was analyzed Western blotting. In addition, expression of PLpro significantly inhibited IMQ-induced activation of interferon-stimulated response element (ISRE) and Nuclear factor-kappaB(NF-κB)promoters. Furthermore, quantitative real time PCR showed that IMQ induced a significant increase of IL-6, IL-8, PKR, and OAS mRNA expression in vector control cells than those in PLpro-expressing cells. Importantly, Western blotting showed that PLpro inhibited the phosphorylation of p38 MAPK, CREB, IRF-3 and STAT1 in IMQ-induced responses. In this study, SARS-CoV PLpro was firstly reported to block TLR7/8 signaling, being helpful for understanding the SARS pathogenesis.
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