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標題: | 小鼠模式中表現無唾液酸神經節甘脂GM1的肝臟佇留CD8 T細胞對HBV的清除扮演必要的角色 Asialo GM1-positive liver-resident CD8 T cells are essential for immune clearance of hepatitis B virus in a mouse model |
作者: | Chi-Chang Sung 宋奇璋 |
指導教授: | 許秉寧(Ping-Ning Hsu) |
關鍵字: | B型肝炎病毒HBV,高壓尾靜脈注射法小鼠模式,無唾液酸神經節甘脂GM1,NFIL3基因缺失小鼠,肝臟CD8 T細胞,去唾液酸糖蛋白受體, Hepatitis B virus,Hydrodynamic injection mouse model,Asialo GM1,Liver resident CD8 T cells,Asialoglycoprotein receptor, |
出版年 : | 2020 |
學位: | 博士 |
摘要: | HBV感染引起與慢性肝炎、肝硬化以及肝癌,造成全球性的健康問題。免疫系統如何控制病毒的機制知之甚少。以前的研究顯示,注射抗AsialoGM1(ASGM1)抗體可阻斷小鼠動物模型中HBV清除的能力,並且一般認為這是由於抗ASGM1導致NK細胞消失。為了進一步驗證NK細胞在HBV清除中的作用,在本研究中,我們使用NFIL3敲除(KO)小鼠(NK細胞缺乏的小鼠)來支持NK細胞對HBV的清除作用。意外的是,在NFIL3 KO小鼠中,清除HBV的能力沒有受到影響,表明NK細胞在HBV清除中不起關鍵作用。此外,用抗ASGM1處理的NFIL3 KO小鼠仍導致抗HBV抗性喪失,暗示存在其他非NK的ASGM1表達免疫細胞參與HBV清除。為了探討這個問題,我們對NFIL3 KO小鼠的肝內ASGM1 +細胞進行了分選,並分析了這些細胞的免疫表型,發現共表達CD44和LFA-1的肝臟駐留CD8 T細胞在這些細胞中佔多數。我們的研究表明, NK細胞在HBV清除中不是必需的。相反地,表達CD44和LFA-1的ASGM1陽性CD8 T細胞是參與抗HBV免疫的主要效應免疫細胞。重要的是,基因轉錄體分析,顯示ASGM1陽性CD8 T細胞相對其他細胞具有獨特的基因轉錄模式,比較像其他研究中所述的組織佇留T細胞(TRM)。我們進一步用過繼性細胞轉移的實驗證明,唯有來自肝臟的ASGM1陽性CD8 T細胞可以順利進入接受小鼠的肝臟中並有接近半數可存活超過二週,證明肝臟的ASGM1陽性CD8 T細胞確實擁有組織佇留的能力。總結以上幾點,我們認為NK細胞在HBV清除中並非關鍵的細胞,相對地,具有組織佇留能力的ASGM1陽性CD8 T細胞,同時表現CD44和LFA-1,在HBV清除中扮演重要的角色。根據本篇的研究,肝細胞上是何種接受子幫助ASGM1的結合,是未來可以進一步的探討的方向。 Hepatitis B virus (HBV) infection causes chronic liver diseases that may progress to chronic hepatitis, liver cirrhosis, and subsequent hepatocellular carcinoma. Previous studies demonstrated that CD8 T cells are critical in HBV elimination. However, whether there is a distinct subtype of CD8 T cells response for the clearance of HBV remains unclear. In this study, we showed that the treatment with anti-asialo GM1(ASGM1) antibody (Ab), an NK cell-depleting Ab for ablating the function of NK cells in vivo, led to impairment of the ability to clear HBV in a mouse animal model. Unexpectedly, the ability to clear HBV was not significantly impaired in NFIL3 KO mice, which are deficient in NK cells, indicating that conventional NK cells do not play a critical role in HBV clearance. Moreover, the ability to clear HBV was abolished when NFIL3 KO mice were further treated with anti-ASGM1 Ab, implying that other non-NK ASGM1-positive immune cells mediate HBV clearance. We isolated intrahepatic ASGM1+ cells from the liver of NFIL3 KO mice and analyzed the immune phenotype of these cells. Our results demonstrated a distinct CD44+ LFA-1hi CD8 T cells that were the major intrahepatic ASGM1-positive immune cells. Importantly, transcriptome analysis revealed that ASGM1-positive CD8 T cells existed substantial differences from others and have similar core gene signature of tissue resident memory cells (TRM). Regarding ASGM1-positive CD8 T cells showed both transcriptional and phenotypic liver residency, we further proved that these cells indeed homed to and remained long-lasting in the liver after adoptive transferring. Collectively, our study indicates that conventional NK cells are not essential for HBV clearance in this mouse model. Instead, ASGM1-positive liver-resident CD8 T cells which express CD44 and LFA-1 are the major effector immune cells mediating anti-HBV immunity. The hepatic receptor which mediates the binding of ASGM1 will be investigated. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/8308 |
DOI: | 10.6342/NTU202002529 |
全文授權: | 同意授權(全球公開) |
顯示於系所單位: | 免疫學研究所 |
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