在核苷（酸）类似物处理下产生的HBV病毒粒子由于不可逆的DN

StephenW

Hepatology. 2019 Jul 6. doi: 10.1002/hep.30844. [Epub ahead of print]
HBV virions produced under nucleos(t)ide analogue treatment are mainly not infectious due to irreversible DNA chain termination.
Liu Y1, Liu H1, Hu Z2, Ding Y3, Pan XB4, Zou J1, Xi J1, Yu G1, Huang H1, Luo MT5, Guo F6, Liu S7, Sheng Q3, Jia J8, Zheng YT5, Chen X1, Guo JT2, Wei L6, Lu F1.
Author information

1
    State Key Laboratory of Natural and Biomimetic Drugs, Department of Microbiology & Infectious Disease Center, School of Basic Medical Sciences, Peking University Health Science Center, Beijing, 100191, P.R. China.
2
    Baruch S. Blumberg Institute, Doylestown, Pennsylvania, 18902, USA.
3
    Department of Infectious Diseases, Shengjing Hospital of China Medical University, Shenyang, 110004, P.R. China.
4
    Hangzhou Key Laboratory of Inflammation and Immunoregulation, Department of Basic Medical Science, School of Medicine, Hangzhou Normal University, Hangzhou, 311121, Zhejiang, P.R. China.
5
    Key Laboratory of Animal Models and Human Disease Mechanisms of the Chinese Academy of Sciences, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming, 650223, China.
6
    Peking University People's Hospital, Peking University Hepatology Institute, Beijing, Key Laboratory of Hepatitis C and Immunotherapy for Liver Diseases, Beijing, 100044, P.R. China.
7
    Beijing Artificial Liver Treatment& Training Center, Beijing Youan Hospital, Capital Medical University, Beijing, 100069, P.R. China.
8
    Liver Research Center, Beijing Friendship Hospital, Capital Medical University, Beijing, 100050, P.R. China.

Abstract

Nucleos(t)ide analogue (NA) have been widely used for the treatment of chronic hepatitis B (CHB). Because viral DNA polymerase lacks proof-reading function (3' exonuclease activity), theoretically, the incorporated NAs would irreversibly terminate viral DNA synthesis. This study explored the natures of nascent HBV DNA and infectivity of progeny virions produced under NA treatment. HBV infectivity was determined by infection of HepG2-NTCP cells and primary human hepatocytes (PHH). The biochemical properties of HBV DNA in the progeny virions were investigated by real-time quantitative polymerase chain reaction (qPCR), Northern blot or Southern blot hybridization, sucrose gradient centrifugation and in vitro endogenous DNA polymerase assay. The progeny HBV virions produced under NA treatment were mainly not infectious to HepG2-NTCP cells or PHH. Biochemical analysis revealed that under NA treatment HBV DNA in nucleaocapsids or virions were predominantly short minus-strand DNA with irreversible termination. This finding was supported by the observation of first disappearance of relaxed circular DNA (rcDNA) and then the proportional decline of HBV DNA levels corresponding to the regions of PreC/C, S and X genes in serial sera of patients receiving NA treatment. Conclusion: HBV virions produced under NA treatment are predominantly replication deficient because the viral genomes are truncated and elongation of DNA chains is irreversibly terminated. Clinically, our results suggest that the viral loads of CHB patients under NA therapy vary with the different regions of genome being detected by qPCR assays. Our findings also imply that NA prevention of perinatal and sexual HBV transmission as well as infection of transplanted livers works not only by reducing viral loads, but also by producing non-infectious virions. This article is protected by copyright. All rights reserved.

This article is protected by copyright. All rights reserved.
KEYWORDS:

Hepatitis B virus; Nucleos(t)ide analogue; infectivity; premature termination

PMID:
    31278760
DOI:
    10.1002/hep.30844

StephenW

肝病。 2019年7月6日doi：10.1002 / hep.30844。 [印刷前的电子版]
在核苷（酸）类似物处理下产生的HBV病毒粒子由于不可逆的DNA链终止而主要不具有感染性。
Liu Y1，Liu H1，Hu Z2，Ding Y3，Pa​​n XB4，Zou J1，Xi J1，Yu G1，Huang H1，Luo MT5，Guo F6，Liu S7，Sheng Q3，Jia J8，Zheng YT5，Chen X1，Guo JT2 ，魏L6，鲁F1。
作者信息

1
    北京大学医学部基础医学院微生物与传染病中心，天然药物及仿生药物国家重点实验室，北京，100191，中国。
2
    Baruch S. Blumberg Institute，Doylestown，Pennsylvania，18902，USA。
3
    中国医科大学附属盛京医院感染科，沉阳110004，中国。
4
    杭州师范大学医学院基础医学系，杭州市炎症与免疫调节重点实验室，浙江杭州，311121，中国。
五
    中国科学院昆明动物研究所中国科学院动物模型与人类疾病机制重点实验室，昆明650223
6
    北京大学人民医院，北京大学肝病研究所，北京，肝炎与肝病免疫治疗重点实验室，北京100044，中国。
7
    首都医科大学附属北京佑安医院北京人工肝治疗培训中心，北京100069，中国。
8
    首都医科大学附属北京友谊医院肝脏研究中心，北京100050，中国。

抽象

核苷（酸）类似物（NA）已被广泛用于治疗慢性乙型肝炎（CHB）。由于病毒DNA聚合酶缺乏校对功能（3'外切核酸酶活性），理论上，纳入的NA会不可逆地终止病毒DNA合成。本研究探讨了新生HBV DNA的性质和NA处理后产生的子代病毒粒子的感染性。通过感染HepG2-NTCP细胞和原代人肝细胞（PHH）确定HBV感染性。通过实时定量聚合酶链反应（qPCR），Northern印迹或Southern印迹杂交，蔗糖梯度离心和体外内源DNA聚合酶测定来研究HBV DNA在子代病毒粒子中的生化特性。在NA处理下产生的子代HBV病毒粒子主要对HepG2-NTCP细胞或PHH没有感染性。生化分析显示，在NA处理下，核衣壳或病毒体中的HBV DNA主要是具有不可逆终止的短负链DNA。这一发现得到了松弛环状DNA（rcDNA）首次消失的观察结果的支持，然后是接受NA治疗的患者系列血清中与PreC / C，S和X基因区域相对应的HBV DNA水平的比例下降。结论：在NA治疗下产生的HBV病毒粒子主要是复制缺陷型，因为病毒基因组被截短并且DNA链的延伸不可逆地终止。临床上，我们的结果表明，在NA治疗下CHB患者的病毒载量随着qPCR测定检测到的基因组的不同区域而变化。我们的研究结果还表明，NA预防围产期和性HBV传播以及移植肝脏的感染不仅可以减少病毒载量，还可以通过产生非感染性病毒粒子。本文受版权保护。版权所有。

本文受版权保护。版权所有。
关键词：

乙型肝炎病毒;核（t）ide类似物;感染;提前终止

结论：
    31278760
DOI：
    10.1002 / hep.30844