<i>In vitro</i> investigation of HBV clinical isolates from Chinese patients reveals that genotype C isolates possess higher infectivity than genotype B isolates

Tingting Liu; Anlei Liu; Yong Liu; Shan Cen; Quan Zhang

doi:10.1016/j.virs.2022.03.008

June 2022

Tingting Liu, Anlei Liu, Yong Liu, Shan Cen and Quan Zhang. In vitro investigation of HBV clinical isolates from Chinese patients reveals that genotype C isolates possess higher infectivity than genotype B isolates[J]. Virologica Sinica, 2022, 37(3): 398-407. doi: 10.1016/j.virs.2022.03.008

Citation: Tingting Liu, Anlei Liu, Yong Liu, Shan Cen, Quan Zhang. In vitro investigation of HBV clinical isolates from Chinese patients reveals that genotype C isolates possess higher infectivity than genotype B isolates .VIROLOGICA SINICA, 2022, 37(3) : 398-407. http://dx.doi.org/10.1016/j.virs.2022.03.008

中国乙型肝炎病毒B基因型和C基因型病毒株感染性的比较

刘婷婷 ^a ,
刘安雷 ^b ,
刘勇 ^c ,
岑山 ^d,, ,
张全 ^c,e,,

1 南京大学医学院附属南京鼓楼医院;
2 中国医学科学院北京协和医学院;
3 中国医学科学院医药生物技术研究所

通讯作者： 岑山, shancen@imb.pumc.edu.cn; 张全, huanlezq44@126.com
收稿日期： 2021-11-03
录用日期： 2022-03-15

摘要

乙型肝炎病毒（Hepatitis B virus，HBV）基因型B和基因型C主要在亚洲地区流行，两者在自然史和临床表现上均有差异。通过基于肝癌细胞系的HBV复制细胞模型，HBV基因型对病毒复制和蛋白表达的影响已在体外得到了大量研究。该模型通过转染的方法将具有复制功能性的HBV基因组导入到肝癌细胞系中，进而可以模拟病毒生活史后期的复制阶段。但是，目前HBV不同基因型对病毒生活史早期感染阶段的影响尚不清楚，主要是因为难于获取体外感染实验所需的高滴度的感染性病毒颗粒。本研究发现通过优化HBV复制细胞模型的转染条件和改造HBV表达载体，可以极大提高病毒颗粒的产量。利用该方法获取高滴度的病毒颗粒进行体外细胞感染实验，我们发现中国C基因型的病毒株感染性高于B基因型。除此之外，我们还鉴定出一个存在于HBV表面抗原小蛋白上的sL21S突变，其可以显著降低C基因型病毒株的感染性，但是并不影响B基因型病毒株的感染性。本研究表明优化后的HBV复制细胞模型可以简单方便的提供各种不同基因组序列的病毒颗粒，便于我们研究大量的不同HBV病毒株的感染性，也将有助于我们对HBV感染不同临床表现机制的理解以及开发和评价靶向HBV生活史早期感染阶段的药物。
- 乙型肝炎病毒
- , B基因型
- , C基因型
- , 临床分离病毒株
- , 感染性
- , sL21S突变

In vitro investigation of HBV clinical isolates from Chinese patients reveals that genotype C isolates possess higher infectivity than genotype B isolates

Tingting Liu ^a ,
Anlei Liu ^b ,
Yong Liu ^c ,
Shan Cen ^d,, ,
Quan Zhang ^c,e,,

a Department of Transfusion Medicine, Nanjing Drum Tower Hospital, The Affiliated Hospital of Nanjing University Medical School, Nanjing, 210008, China;
b Department of Emergency Medicine, Peking Union Medical College Hospital, Chinese Academy of Medical Science, Beijing, 100730, China;
c Department of Laboratory Medicine, Nanjing Drum Tower Hospital and Jiangsu Key Laboratory for Molecular Medicine, The Affiliated Hospital of Nanjing University Medical School, Nanjing, 210008, China;

Corresponding author: Shan Cen, shancen@imb.pumc.edu.cn Quan Zhang, huanlezq44@126.com
Received Date: 03 November 2021
Accepted Date: 15 March 2022

Abstract

Hepatitis B virus (HBV) genotype B and C are two major genotypes that are prevalent in Asia and differ in natural history and disease progression. The impact of HBV genotypes on viral replication and protein expression has been explored by the transfection of hepatoma cells with replication-competent HBV DNA, which mimics the later stages of the viral life cycle. However, the influence of HBV genotypes on the early events of viral infection remains undetermined, mainly due to the difficulties in obtaining sufficient infectious viral particles for infection assays. Here, we report that a high-titer HBV inoculum can be generated from the transient transfection-based cell model after optimizing transfection conditions and modifying the HBV-expressing construct. By performing in vitro infection assays using transiently transfected derived viruses, we found that clinical genotype C isolates possessed higher infectivity than genotype B isolates. Moreover, we identified a naturally occurring mutation sL21S in small hepatitis B surface protein, which markedly decreased the infectivity of HBV genotype C isolates, but not that of genotype B isolates. In summary, using infectious viral particles provided by the optimized transient transfection-based cell model, we have been able to investigate a wide range of HBV variants on viral infectivity, which may contribute to our understanding of the reasons for different clinical outcomes in HBV infections and the development of therapeutic drugs targeting the early stages of HBV life cycle.
- Hepatitis B virus (HBV)
- , Genotype B
- , Genotype C
- , Clinical isolates
- , Infectivity
- , sL21S

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Electronic Supplementary Material

10.1016j.virs.2022.03.008-ESM.docx