细胞外HBV RNA的长度是异质的，并且除了慢性乙型肝炎患者中

StephenW

J Virol. 2018 Oct 3. pii: JVI.00798-18. doi: 10.1128/JVI.00798-18. [Epub ahead of print]
Extracellular HBV RNAs are heterogeneous in length and circulate as capsid-antibody-complexes in addition to virions in chronic hepatitis B patients.
Bai L1,2, Zhang X3, Li W1, Wu M1, Liu J2, Kozlowski M1,2, Chen L1, Zhang J4, Huang Y1,4, Yuan Z3,2.
Author information

1
    Shanghai Public Health Clinical Center, Fudan University, Shanghai, China.
2
    Key Laboratory of Medical Molecular Virology, Ministry of Education and Health, School of Basic Medical Sciences, Shanghai Medical College of Fudan University, Shanghai, China.
3
    Shanghai Public Health Clinical Center, Fudan University, Shanghai, China. [email protected] [email protected].
4
    Department of Infectious Diseases, Huashan Hospital, Fudan University, Shanghai, China.

Abstract

Extracellular HBV RNA has been detected in both HBV-replicating cell culture media and sera from chronic hepatitis B (CHB) patients, but its exact origin and composition remain controversial. Here, we demonstrated that extracellular HBV RNA species were of heterogeneous lengths, ranging from the length of pregenomic RNA to a few hundred nucleotides. In cell models, these RNAs were predominantly associated with naked capsids although virions also harbored a minority of them. Moreover, HBV RNAs in hepatitis B patients' blood circulation were localized in unenveloped capsids in the form of capsid-antibody-complexes (CACs) and in virions. Furthermore, we showed that extracellular HBV RNAs could serve as template for viral DNA synthesis. In conclusion, extracellular HBV RNAs mainly consist of pgRNA or the pgRNA species degraded by the RNase H domain of the polymerase in the process of viral DNA synthesis and circulate as CACs and virions. Their presence in the blood circulation of CHB patients may be exploited to develop novel biomarkers for HBV persistence.Importance Although increasing evidence suggests the presence of extracellular HBV RNA species, their origin and molecular forms are still under debate. In addition to the infectious virions, HBV is known to secrete several species of incomplete viral particles, including hepatitis B surface antigen (HBsAg) particles, naked capsids and empty virions during its replication cycle. Here, we demonstrated that extracellular HBV RNAs were associated with naked capsids and virions in HepAD38 cells. Interestingly, we found that unenveloped capsids circulate in the blood of hepatitis B patients in the form of capsids-antibody-complexes (CACs) and, together with virions, serve as vehicles carrying these RNAs molecules. Moreover, extracellular HBV RNAs are heterogeneous in length and represent either pregenomic RNA (pgRNA) or products of incomplete reverse transcription during viral replication. These findings provide a conceptual basis for further application of extracellular RNA species as novel biomarkers for HBV persistence.

PMID:
    30282709
DOI:
    10.1128/JVI.00798-18

StephenW

J Virol。 2018年10月3日.pii：JVI.00798-18。 doi：10.1128 / JVI.00798-18。 [提前打印]
细胞外HBV RNA的长度是异质的，并且除了慢性乙型肝炎患者中的病毒粒子外，还作为衣壳 - 抗体 - 复合物循环。
Bai L1,2，Zhang X3，Li W1，Wu M1，Liu J2，Kozlowski M1,2，Chen L1，Zhang J4，Huang Y1,4，Yuan Z3,2。
作者信息

1
    复旦大学上海市公共卫生临床中心，上海
2
    复旦大学上海医学院基础医学院教育与卫生部医学分子病毒学重点实验室，上海
3
    复旦大学上海市公共卫生临床中心，上海[email protected] [email protected]。
4
    复旦大学附属华山医院感染科，上海

抽象

已经在HBV复制细胞培养基和慢性乙型肝炎（CHB）患者的血清中检测到细胞外HBV RNA，但其确切起源和组成仍存在争议。在这里，我们证明了细胞外HBV RNA种类具有不同的长度，范围从前基因组RNA的长度到几百个核苷酸。在细胞模型中，这些RNA主要与裸衣壳相关，尽管病毒粒子也含有少数这些RNA。此外，乙型肝炎患者血液循环中的HBV RNA以衣壳 - 抗体 - 复合物（CAC）和病毒粒子的形式定位于未发育的衣壳中。此外，我们发现细胞外HBV RNA可以作为病毒DNA合成的模板。总之，细胞外HBV RNA主要由pgRNA或在病毒DNA合成过程中被聚合酶的RNA酶H结构域降解的pgRNA物质组成，并作为CAC和病毒体循环。它们在CHB患者的血液循环中的存在可被用于开发用于HBV持久性的新型生物标志物。重要性尽管越来越多的证据表明存在细胞外HBV RNA种类，但它们的起源和分子形式仍然存在争议。除了感染性病毒粒子外，已知HBV在其复制周期中分泌几种不完整的病毒颗粒，包括乙型肝炎表面抗原（HBsAg）颗粒，裸衣壳和空病毒粒子。在这里，我们证明细胞外HBV RNA与HepAD38细胞中的裸衣壳和病毒体相关。有趣的是，我们发现未发育的衣壳以衣壳 - 抗体 - 复合物（CAC）的形式在乙型肝炎患者的血液中循环，并且与病毒粒子一起作为携带这些RNA分子的载体。此外，细胞外HBV RNA的长度是异质的，并且代表在病毒复制期间的前基因组RNA（pgRNA）或不完全逆转录的产物。这些发现为进一步应用细胞外RNA种类作为HBV持久性的新型生物标志物提供了概念基础。

结论：
    30282709
DOI：
    10.1128 / JVI.00798-18

StephenW

https://jvi.asm.org/content/jvi/ ... I.00798-18.full.pdf

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