HBV 复制肝细胞的体内生物发光成像允许监测抗病毒免疫

StephenW

HBV 复制肝细胞的体内生物发光成像允许监测抗病毒免疫
Katrin Manske 1，Annika Schneider 1，Chunkyu Ko 2 3，Percy A Knolle 1 4，Katja Steiger 5，Ulrike Protzer 2 4，Dirk Wohlleber 1
隶属关系
隶属关系

    1
    分子免疫学研究所，Klinikum Rechts der Isar，TUM 医学院，慕尼黑工业大学，80333 慕尼黑，德国。
    2
    慕尼黑工业大学病毒学研究所，德国慕尼黑 81675。
    3
    韩国化学技术研究所 (KRICT) 治疗与生物技术部传染病治疗研究中心，韩国大田 34114。
    4
    德国感染研究中心 (DZIF)，慕尼黑合作伙伴站点，81675 慕尼黑，德国。
    5
    慕尼黑工业大学病理学研究所，德国慕尼黑 81675。

    PMID：34835079 DOI：10.3390/v13112273

抽象的

对乙型肝炎病毒 (HBV) 感染的免疫是复杂的，目前尚未完全了解，包括导致慢性乙型肝炎发展的决定性因素。合适的动物模型。在这里，我们描述了表达与荧光素酶 (Ad-HBV-Luc) 相关联的 HBV 1.3 超长基因组的重组腺病毒的产生，从而可以精确分析受感染肝细胞的数量。这使得能够对 HBV 特异性 CD8 T 细胞和小鼠抗病毒免疫的发生进行敏感和密切的监测。高剂量的 Ad-HBV-Luc 发展为慢性乙型肝炎，伴有功能失调的 CD8 T 细胞，其特征是 PD1 和 TOX 高表达，KLRG1 和 GzmB 低表达。相比之下，低剂量的 Ad-HBV-Luc 感染导致急性肝炎，CD8 T 细胞介导的 HBV 复制肝细胞的清除与升高的 sALT 水平和细胞毒性 HBV 特异性 CD8 T 细胞数量增加有关。因此，感染剂量是诱导小鼠急性自限性或慢性 HBV 感染的关键因素。总之，新的 Ad-HBV-Luc 载体将允许在急性和慢性感染期间对 HBV 特异性免疫反应进行高度敏感和时间分辨的分析。

关键词：CD8 T细胞；乙肝病毒； T 细胞功能障碍;生物发光成像（BLI）；免疫;肝脏免疫学。
拨款支持

    项目编号 272983813 - TRR179/Deutsche Forschungsgemeinschaft

StephenW

In Vivo Bioluminescence Imaging of HBV Replicating Hepatocytes Allows for the Monitoring of Anti-Viral Immunity
Katrin Manske  1 , Annika Schneider  1 , Chunkyu Ko  2   3 , Percy A Knolle  1   4 , Katja Steiger  5 , Ulrike Protzer  2   4 , Dirk Wohlleber  1
Affiliations
Affiliations

    1
    Institute of Molecular Immunology, Klinikum Rechts der Isar, TUM School of Medicine, Technical University of Munich, 80333 Munich, Germany.
    2
    Institute of Virology, Technical University of Munich, 81675 Munich, Germany.
    3
    Infectious Diseases Therapeutic Research Center, Therapeutics & Biotechnology Division, Korea Research Institute of Chemical Technology (KRICT), Daejeon 34114, Korea.
    4
    German Center for Infection Research (DZIF), Munich Partner Site, 81675 Munich, Germany.
    5
    Institute of Pathology, Technical University of Munich, 81675 Munich, Germany.

    PMID: 34835079 DOI: 10.3390/v13112273

Abstract

Immunity against hepatitis B virus (HBV) infection is complex and not entirely understood so far, including the decisive factors leading to the development of chronic hepatitis B. This lack of a mechanistic understanding of HBV-specific immunity is also caused by a limited number of suitable animal models. Here, we describe the generation of a recombinant adenovirus expressing an HBV 1.3-overlength genome linked to luciferase (Ad-HBV-Luc) allowing for precise analysis of the quantity of infected hepatocytes. This enables sensitive and close-meshed monitoring of HBV-specific CD8 T cells and the onset of anti-viral immunity in mice. A high dose of Ad-HBV-Luc developed into chronic hepatitis B accompanied by dysfunctional CD8 T cells characterized by high expression of PD1 and TOX and low expression of KLRG1 and GzmB. In contrast, a low dose of Ad-HBV-Luc infection resulted in acute hepatitis with CD8 T cell-mediated elimination of HBV-replicating hepatocytes associated with elevated sALT levels and increased numbers of cytotoxic HBV-specific CD8 T cells. Thus, the infectious dose was a critical factor to induce either acute self-limited or chronic HBV infection in mice. Taken together, the new Ad-HBV-Luc vector will allow for highly sensitive and time-resolved analysis of HBV-specific immune responses during acute and chronic infection.

Keywords: CD8 T cells; HBV; T cell dysfunction; bioluminescence imaging (BLI); immunity; liver immunology.
Grant support

    Projektnummer 272983813 - TRR179/Deutsche Forschungsgemeinschaft