从慢性HBV携带者收获的感染组织中来自整合子的病毒RNA的相

StephenW

J Virol. 2018 Feb 28. pii: JVI.02221-17. doi: 10.1128/JVI.02221-17. [Epub ahead of print]
Relative abundance of the integrant-derived viral RNAs in infected tissues harvested from chronic HBV carriers.Freitas N1, Lukash T1, Gunewardena S2, Chappell B1, Slagle BL3, Gudima SO4.
Author information1Department of Microbiology, Molecular Genetics and Immunology, University of Kansas Medical Center, Kansas City, Kansas, USA.2Department of Molecular and Integrative Physiology, University of Kansas Medical Center, Kansas City, Kansas, USA.3Department of Molecular Virology and Microbiology, Baylor College of Medicine, Houston, Texas, USA.4Department of Microbiology, Molecular Genetics and Immunology, University of Kansas Medical Center, Kansas City, Kansas, USA [email protected].

AbstractFive matching sets of non-malignant liver tissues and HCCs from individuals chronically infected with hepatitis B virus (HBV) were examined. The HBV genomic sequences were determined using overlapping PCR amplicons covering the entire viral genome. Four pairs of tissues were infected with HBV of genotype C, while one pair - with genotype B. HBV replication markers were found in all tissues. In majority of HCC samples, the levels of pre-genomic/pre-core RNA (pgRNA) and covalently closed circular DNA (cccDNA) were lower than those of liver tissue counterparts. Regardless of the presence of HBV replication markers, (i) integrant-derived HBV RNAs (id-RNAs) were found using RT-PCR analysis in all tissues, and were considerably abundant or predominant in 6/10 tissue samples (2 livers and 4 HCCs); (ii) the RNAs that were polyadenylated using cryptic HBV polyadenylation signal and therefore could be produced by HBV replication or derived from integrated HBV DNA were found in 5/10 samples (3 livers and 2 HCCs), and were considerably abundant species in 3/10 tissues (2 livers and 1 HCC); and (iii) cccDNA-transcribed RNAs polyadenylated near position 1931 were not abundant in 7/10 tissues (2 livers and 5 HCCs), and were predominant only in two livers. Subsequent RNA sequencing analysis of selected liver/HCC samples also showed relative abundance of id-RNAs in most of examined tissues. Our findings suggesting that id-RNAs could represent a significant source of HBV envelope proteins, which is independent of viral replication, are discussed in the context of possible contribution of id-RNAs to the HBV life cycle.IMPORTANCE The relative abundance of integrant-derived HBV RNAs (id-RNAs) in chronically infected tissues suggests that id-RNAs coding for the envelope proteins may facilitate production of considerable fraction of surface antigens (HBsAg) in infected cells bearing HBV integrants. If the same cells support HBV replication, then a significant fraction of assembled HBV virions could bear id-RNA-derived HBsAg as a major component of their envelopes. Therefore, infectivity of these HBV virions, and their ability to facilitate virus cell-to-cell spread could be determined mainly by the properties of id-RNA-derived envelope proteins, and not by the properties of replication-derived HBsAg. These interpretations suggest that id-RNAs may play a role in maintenance of chronic HBV infection, and therefore contribute to HBV life cycle. Furthermore, the production of HBsAg from id-RNAs independently of viral replication may at least in part explain why treatment with interferon or nucleos(t)ides in most cases failed to achieve loss of serum HBsAg.
PMID:29491161DOI:10.1128/JVI.02221-17

StephenW

J Virol。 2018年2月28日。pii：JVI.02221-17。 doi：10.1128 / JVI.02221-17。 [电子版提前打印]
从慢性HBV携带者收获的感染组织中来自整合子的病毒RNA的相对丰度。
Freitas N1，Lukash T1，Gunewardena S2，Chappell B1，Slagle BL3，Gudima SO4。
作者信息

1
    美国堪萨斯州堪萨斯城堪萨斯大学医学中心微生物学分子遗传与免疫学系。
2
    美国堪萨斯州堪萨斯城堪萨斯大学医学中心分子与整合生理学系。
3
    贝勒医学院分子病毒学和微生物学系，美国得克萨斯州休斯顿。
4
    美国堪萨斯州堪萨斯城堪萨斯大学医学中心微生物学分子遗传与免疫学系[email protected]。

抽象

检查了5组匹配的非恶性肝组织和来自慢性感染乙型肝炎病毒（HBV）的个体的HCC。使用覆盖整个病毒基因组的重叠PCR扩增子确定HBV基因组序列。 HBV基因型C感染四对组织，一对 - 基因型B，HBV复制标志物在所有组织中均有发现。在大多数HCC样品中，前基因组/前核心RNA（pgRNA）和共价闭合环状DNA（cccDNA）的水平低于肝组织对应物的水平。无论HBV复制标记物是否存在，（i）在所有组织中使用RT-PCR分析发现了来源于整合子的HBV RNA（id-RNA），并且在6/10组织样品（2个肝脏和4个肝癌）; （ii）在5/10个样品（3个肝脏和2个HCC）中发现使用隐藏的HBV多聚腺苷酸化信号聚腺苷酸化并因此可以由HBV复制产生或从整合的HBV DNA衍生的RNAs，并且在3 / 10个组织（2个肝脏和1个HCC）;和（iii）靠近1931位多腺苷酸化的cccDNA转录的RNA在7/10组织（2个肝脏和5个HCC）中不丰富，并且仅在两个肝脏中占优势。选择的肝/ HCC样品的随后的RNA测序分析也显示在大多数检查的组织中id-RNA的相对丰度。我们的研究结果表明，id-RNAs可能代表HBV包膜蛋白的一个重要来源，它与病毒复制无关，在id-RNAs可能对HBV生命周期产生影响的背景下进行了讨论。IMPORT立体成分的相对丰度慢性感染组织中的HBV RNAs（id-RNAs）表明编码包膜蛋白的id-RNAs可能有助于在带有HBV整合子的感染细胞中产生大量表面抗原（HBsAg）。如果相同的细胞支持HBV复制，则组装的HBV病毒颗粒的显着部分可以承担由id-RNA衍生的HBsAg作为其信封的主要组分。因此，这些HBV病毒粒子的感染性以及它们促进病毒细胞间传播的能力可主要由id-RNA衍生的包膜蛋白的性质决定，而不是由复制衍生的HBsAg的性质决定。这些解释表明，id-RNAs可能在维持慢性HBV感染中起作用，因此有助于HBV的生命周期。此外，独立于病毒复制的来自id-RNA的HBsAg的产生可能至少部分解释了为什么在大多数情况下用干扰素或核苷（t）治疗未能达到血清HBsAg的损失。