乙型肝炎病毒前C区/核心基因羧基端突变对病毒生物合成与宿

StephenW

The Impact of Hepatitis B Virus Precore/Core Gene Carboxyl Terminal Mutations on Viral Biosynthesis and the Host Immune Response

    Jia-Feng Wu1,
    Yen-Hsuan Ni1,2,
    Huey-Ling Chen1,3,
    Hong-Yuan Hsu1 and
    Mei-Hwei Chang1,3

+ Author Affiliations

    1Department of Pediatrics, National Taiwan University Hospital, Taipei, Taiwan
    2Department of Genetics, National Taiwan University Hospital, Taipei, Taiwan
    3Department of Hepatitis Research Center, National Taiwan University Hospital, Taipei, Taiwan

    Correspondence: Mei-Hwei Chang, MD., Department of Pediatrics and Hepatitis Research Center, National Taiwan University Hospital; No. 8, Chung-Shan South. Rd., Taipei, Taiwan. TEL: (886-2)-23123456, ext. 71723, FAX: (886-2)-23938871; E-mail: [email protected]

Abstract

Background. We aimed to elucidate the impact of hepatitis B virus (HBV) precore/core gene mutations on spontaneous hepatitis B e antigen (HBeAg)-seroconversion, HBV biosynthesis, and the human immune responses in chronic HBV-infected patients.

Methods. We analyzed the HBV precore/core gene sequences by cloning method in 33 chronic HBV-infected patients during the inflammatory phase before spontaneous HBeAg-seroconversion. The impact of the most prevalent mutant on HBeAg biosynthesis was assessed by Western blot and native agarose gel analysis in Huh7 cells, and the human immune responses were assessed by in vitro stimulation of CD3+CD8+ T lymphocytes of chronic HBV-infected subjects.

Results. The P135Q and G1896A were the most prevalent mutants before HBeAg-seroconversion, acting as markers of HBeAg-seroconversion (hazard ratios=2.75 and 4.50; P=0.01 and <0.001, respectively). The P135Q mutants displayed decreased HBeAg secretion and HBV capsid molecular-weight, while showing increased 22kD HBeAg pro-protein accumulation in Huh7 cells. The P135Q mutant peptide induced less interferon-γ expression in CD3+CD8+ T lymphocytes in HBeAg-negative subjects as compared to the wild type peptide (P=0.03).

Conclusions. The HBV P135Q mutant emergence during the inflammatory phase was associated with HBeAg-seroconversion. It was associated with altered HBV capsid assembly, HBeAg biosynthesis, and reduced human immune responses following HBeAg-seroconversion.

    Received August 22, 2013.
    Revision received October 4, 2013.
    Accepted October 11, 2013.

StephenW

乙型肝炎病毒前C区/核心基因羧基端突变对病毒生物合成与宿主的免疫反应的影响

    贾锋武1 ，
    颜轩妮， 2 ，
    惠凌晨1 ， 3 ，
    宏远Hsu1和
    美慧Chang1 ， 3

+作者所属机构

    儿科，台大医院，台北，台湾教研室
    遗传学教研室中，台大医院，台北，台湾
    3Department肝炎研究中心，台大医院，台北，台湾

    通讯：张美惠，医学博士，儿科和肝炎研究中心，国立台湾大学医学院附设医院系; 8号，中山南。路，台北，台湾。电话：（886-2 ） -23123456 ，分机。 71723 ，传真： （ 886-2 ） -23938871 ;电子邮箱： [email protected]

摘要

背景。我们的目的是阐明乙型肝炎病毒（HBV ）前C / C基因变异对自发乙型肝炎e抗原（HBeAg ）血清转换， HBV生物合成，并在慢性HBV感染者的人体免疫反应的影响。

方法。我们通过在自发性HBeAg血清转换前炎症期克隆方法在33个慢性HBV感染者分析HBV前C区/核心基因序列。对HBeAg合成最普遍突变的影响，用Western blot和在Huh7细胞原生琼脂糖凝胶电泳分析评估，以及人体免疫反应进行体外刺激慢性HBV感染者的CD3 + CD8 + T淋巴细胞的评估。

结果。该P135Q和G1896A为HBeAg血清转换之前，最常见的突变体，充当的HBeAg血清转换的标记（危险比= 2.75和4.50 ，P = 0.01和< 0.001） 。显示P135Q突变体降低HBeAg的分泌及HBV衣壳分子量，而呈现增加的22KD大三阳亲蛋白质积累在Huh7细胞。该P135Q突变肽诱导的CD3 + CD8 + T淋巴细胞较少干扰素- γ表达HBeAg阴性受试者相比于野生型肽（P = 0.03）。

结论。在炎症期的乙肝P135Q突变的出现是与e抗原血清学转换有关。它与乙肝改变衣壳组装， HBeAg的生物合成，并遵循的HBeAg血清学转换降低人体的免疫反应有关。

    收到2013年8月22日。
    修订收到2013年10月4日。
    接受2013年10月11日。