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標題: | 探討NS5蛋白入核對登革病毒在埃及斑蚊體內複製之重要性 Dengue virus NS5 protein nuclear localization is crucial for virus replication in Aedes aegypti |
作者: | Yi-Shan Wu 吳依珊 |
指導教授: | 蕭信宏 |
關鍵字: | 埃及斑蚊,登革病毒,入核,免疫調控,病毒複製, Aedes aegypti,dengue virus,nuclear localization,immune regulation,virus replication, |
出版年 : | 2019 |
學位: | 碩士 |
摘要: | 蚊子是許多疾病的傳播媒介,例如:瘧疾、登革熱,茲卡、屈躬病、日本腦炎、西尼羅河熱以及黃熱病等。其中,登革熱為目前重要的疾病之一,根據世界衛生組織統計,全球每年大約有3億9千萬的感染病例,且當中約9千萬人具有嚴重臨床症狀。而在2014至2015年間台灣共有超過5萬件的感染案例,但由於目前仍無有效的治療藥物上市,因此,如何預防病媒蚊叮咬以及控制病媒蚊的數量,是目前防治蚊媒傳染疾病最重要的研究課題。根據先前哺乳類研究文獻指出,第二型登革病毒 (dengue virus serotype 2, DENV2)中的非結構性蛋白5 (non-structural protein 5, NS5)在感染宿主細胞的過程中,會進入宿主細胞的細胞核內,且會對病毒的感染力產生影響,因此,本研究以埃及斑蚊為研究模式,探討NS5蛋白入核對登革病毒在埃及斑蚊體內複製之重要性。首先,我們利用免疫螢光染色法分析登革病毒第一至四型的NS5蛋白入核狀況,結果顯示只有第二型登革病毒的NS5蛋白在埃及斑蚊體內會進入細胞核中。接著,我們利用可抑制NS5蛋白入核的抑制劑Ivermectin來處理埃及斑蚊,發現經Ivermectin處理的組別,登革病毒的複製能力及感染力皆有顯著下降。除此之外,先前研究顯示NS5蛋白入核會影響messenger RNA的剪接,而在本研究中也發現NS5蛋白入核會影響免疫相關基因Caspar的剪接。未來我們將更進一步探討NS5蛋白入核對於影響登革病毒複製能力的詳細機制,以及探討與Caspar相關的免疫調控機制。 Mosquitoes are one of the fatal animals in the world and they act as vectors for several diseases, including malaria, dengue fever, Zika, japanese encephalitis, West Nile fever, and chikungunya. Among these diseases, dengue fever is currently one of the world’s most important tropical diseases. Previous studies have indicated that the non-structural protein 5 (NS5) was able to translocate into the nucleus and influence the dengue virus serotype 2 (DENV2) infectivity in mammalian cell lines. However, the detail mechanisms of NS5-mediated infectivity remain largely unknown. Therefore, the aim of this study is to investigate the role of the dengue virus NS5 nuclear localization on DENV replication in the mosquito Aedes aegypti. We showed that dengue virus NS5 was localized in the nucleus only in the case of DENV2 (16681 strain) infection in Aedes aegypti. Next, we investigated the effect of NS5 nuclear localization in viral replication by application of Ivermectin, which was demonstrated to be an inhibitor for NS5 protein nuclear localization. We showed that treatment of Ivermectin resulted in the significant reduction of DENV2 replication and infectivity. We also showed that Caspar splicing was inhibited in the case of NS5 nuclear localization, suggesting the important role of NS5 nuclear localization on host mRNA splicing. In the future, we will further investigate the detail mechanisms underlying the regulation of NS5 nuclear localization on DENV2 replication and immune response modulation. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/74172 |
DOI: | 10.6342/NTU201903186 |
全文授權: | 有償授權 |
顯示於系所單位: | 微生物學科所 |
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