利用永生化人類肝細胞建立新型乙肝病毒培養系統

StephenW

Establishment of a novel hepatitis B virus culture system using immortalized human hepatocytes
Yuichi Akahori  1   2 , Hiroki Kato  1   2 , Takashi Fujita  1   2 , Kohji Moriishi  3 , Yasuhito Tanaka  4 , Koichi Watashi  5 , Michio Imamura  6 , Kazuaki Chayama  6 , Takaji Wakita  5 , Makoto Hijikata  7   8
Affiliations
Affiliations

    1
    Institute for Frontier Life and Medical Sciences, Kyoto University, Kyoto, Japan.
    2
    Graduate School of Biostudies, Kyoto University, Kyoto, Japan.
    3
    Interdisciplinary Graduate School of Medicine and Engineering, University of Yamanashi, Kofu, Japan.
    4
    Graduate School of Medical Sciences, Nagoya City University, Nagoya, Japan.
    5
    Department of Virology II, National Institute of Infectious Diseases, Tokyo, Japan.
    6
    Department of Gastroenterology and Metabolism, Hiroshima University Hospital, Hiroshima, Japan.
    7
    Institute for Frontier Life and Medical Sciences, Kyoto University, Kyoto, Japan. [email protected].
    8
    Graduate School of Biostudies, Kyoto University, Kyoto, Japan. [email protected].

    PMID: 33303813 DOI: 10.1038/s41598-020-78655-x

Free article
Abstract

Recent development of hepatitis B virus (HBV) culture systems has made it possible to analyze the almost all steps of the viral life cycle. However, the reproducibility of interaction between HBV and host cells seemed inaccurate in those systems because of utilization of cancer cell lines with a difference from hepatocytes in the majority of cases. In this study, in order to resolve this point, a novel HBV culture system using non-cancer-derived immortalized human hepatocytes derived cell lines, producing exogenous human sodium taurocholate cotransporting polypeptide, was developed. One of the cell clones, E/NtG8 cells, was permissive to both blood-borne HBV (HBVbb) and culture-derived recombinant HBV when cultured in the three-dimensional condition. Furthermore, the production of infectious HBV particles, which showed the similar physicochemical properties to HBVbb, was observed for about a month after HBVbb infection in this system, suggesting that it may reproduce whole steps of the HBV lifecycle under the condition analogous to human liver cells infected with HBV. This system seemed to contribute not only to find novel interactions between HBV and host cells but also to understand mechanism of HBV pathogenesis.

StephenW

利用永生化人類肝細胞建立新型乙肝病毒培養系統
赤森雄一1 2，加藤裕樹1 2，藤田隆1 2，森田公治3，田中康人4，晃一郎5，今村道夫6，茶山一明6，秋田孝司5，日方誠7 8
隸屬關係
隸屬關係

    1個
    日本京都大學前沿生命與醫學研究所。
    2
    日本京都大學生物研究科研究生院。
    3
    日本山梨大學山梨大學醫學與工程學交叉學科研究生院。
    4
    日本名古屋市名古屋市立大學醫學研究科。
    5
    日本東京國立傳染病研究所病毒學系II。
    6
    日本廣島大學附屬醫院消化內科。
    7
    日本京都大學前沿生命與醫學研究所。 [email protected]。
    8
    日本京都大學生物研究科研究生院。 [email protected]。

    PMID：33303813 DOI：10.1038 / s41598-020-78655-x

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抽象

乙型肝炎病毒（HBV）培養系統的最新發展使得分析病毒生命週期的幾乎所有步驟成為可能。但是，在大多數係統中，由於利用癌細胞係與肝細胞有所不同，因此在這些系統中，HBV與宿主細胞之間相互作用的再現性似乎不准確。在本研究中，為了解決這一問題，開發了一種使用非癌症來源的永生化人肝細胞衍生細胞系的新型HBV培養系統，該系統可生產外源性人牛磺膽酸鈉共轉運多肽。當在三維條件下培養時，其中一種細胞克隆E / NtG8細胞可同時攜帶血源性HBV（HBVbb）和培養物衍生的重組HBV。此外，在該系統中感染HBVbb後約一個月，觀察到了具有與HBVbb相似的理化特性的傳染性HBV顆粒的產生，這表明它可以在類似於人肝細胞的條件下重現HBV生命週期的整個步驟。感染了乙肝病毒。該系統似乎不僅有助於發現HBV與宿主細胞之間的新型相互作用，而且有助於了解HBV發病機理。

StephenW

https://www.nature.com/articles/s41598-020-78655-x.pdf