[推荐]累积的病毒进化的改变在乙肝e抗原血

StephenW

Gut 2013;62:1347-1355 doi:10.1136/gutjnl-2012-302408

    Hepatology

    Original article

Cumulative viral evolutionary changes in chronic hepatitis B virus infection precedes hepatitis B e antigen seroconversion

    Yan Cheng1,
    Stephane Guindon2,
    Allen Rodrigo3,
    Lin Ying Wee2,
    Masafumi Inoue4,
    Alex J V Thompson5,
    Stephen Locarnini5,
    Seng Gee Lim1,6

- Author Affiliations

    1Department of Medicine, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, Singapore
    2Department of Statistics, University of Auckland, Auckland, New Zealand
    3Department of Biology, National Evolutionary Synthesis Center, Duke University, Durham, North Carolina, USA
    4Experimental Therapeutic Center, Agency for Science, Technology and Research, Singapore
    5Research and Molecular Development, Victorian Infectious Diseases Reference Laboratory, Monash University, Melbourne, Australia
    6Department of Gastroenterology and Hepatology, National University Health System, Singapore, Singapore

    Correspondence to Professor Seng Gee Lim, Department of Gastroenterology and Hepatology, National University Health System, Tower Block level 10, 1E Kent Ridge Road, Singapore 119228, Singapore; [email protected]

    Received 27 February 2012
    Revised 16 November 2012
    Accepted 22 November 2012
    Published Online First 15 December 2012

Abstract

Objective To examine viral evolutionary changes and their relationship to hepatitis B e antigen (HBeAg) seroconversion.

Design A matched case–control study of HBeAg seroconverters (n=8) and non-seroconverters (n=7) with adequate stored sera before seroconversion was performed. Nested PCR, cloning and sequencing of hepatitis B virus (HBV) precore/core gene was performed. Sequences were aligned using Clustal X2.0, followed by construction of phylogenetic trees using Pebble 1.0. Viral diversity, evolutionary rates and positive selection were then analysed.

Results Baseline HBV quasispecies viral diversity was identical in seroconverters and non-seroconverters 10 years before seroconversion but started to increase approximately 3 years later. Concurrently, precore stop codon (PSC) mutations appeared. Some 2 years later, HBV-DNA declined, together with a dramatic reduction in HBeAg titres. Just before HBeAg seroconversion, seroconverters had HBV-DNA levels 2 log lower (p=0.008), HBeAg titres 310-fold smaller (p=0.02), PSC mutations > 25% (p<0.001), viral evolution 8.1-fold higher (p=0.01) and viral diversity 2.9-fold higher (p<0.001), compared to non-seroconverters, with a 9.3-fold higher viral diversity than baseline (p=0.011). Phylogenetic trees in seroconverters showed clustering of separate time points and longer branch lengths than non-seroconverters (p=0.01). Positive selection was detected in five of eight seroconverters but none in non-seroconverters (p=0.026). There was significant negative correlation between viral diversity (rs=−0.60, p<0.001) and HBV-DNA or HBeAg (rs=−0.58, p=0.006) levels; and positive correlation with PSC mutations (rs=0.38, p=0.009). Over time, the significant positive correlation was viral diversity (rs=0.65, p<0.001), while negative correlation was HBV-DNA (rs=−0.627, p<0.001) and HBeAg levels (rs=−0.512, p=0.015).

Conclusions Cumulative viral evolutionary changes that precede HBeAg seroconversion provide insights into this event that may have implications for therapy.

StephenW

累积的病毒进化的改变在慢性乙肝病毒感染者，乙肝e抗原血清学转换之前

    阎成1，
    斯特凡Guindon2，
    艾伦Rodrigo3，
    林英Wee2，
    雅文Inoue4
    亚历克斯J V Thompson5
    斯蒂芬Locarnini5
    恒生啧啧LIM1 6

- 作者所属机构

    1，新加坡国立大学杨潞龄医学院，新加坡，新加坡医学院
    2统计，奥克兰大学，奥克兰，新西兰
    3生物学国家进化综合中心，美国杜克大学，北卡罗莱纳州，
    治疗中心4Experimental，机构科学，技术和研究，新加坡
    5Research与分子发展，维多利亚传染病参考实验室，澳大利亚墨尔本莫纳什大学，
    新加坡，新加坡国立大学健康系统，胃肠病学和肝病学6Department

    函授生教授志廉，胃肠病学和肝病学系，国立大学健康系统，10座级，1E肯特岗路，新加坡119228，新加坡; [email protected]

    2012年2月27日
    2012年11月16日修订
    2012年11月22日
    在线发布2012年12月前15

抽象

目的研究病毒进化的改变和他们的关系，乙肝e抗原（HBeAg）血清学转换。

设计一个匹配的病例对照研究的HBeAg血清转化组（n = 8）和非组（n = 7）提供足够的存储血清血清转化前进行血清转换。巢式PCR，B型肝炎病毒（HBV）前C / C基因的克隆和测序。使用用Clustal X2.0，建设使用卵石1.0的系统发育树序列一致。病毒的多样性，进化速率和积极的选择，然后进行分析。

结果基线HBV准种病毒的多样性是相同的血清阳转和非血清转化的前10年血清转换，但大约3年后开始增加。同时，前C区终止密码子（PSC）的突变出现了。大约2年后，HBV-DNA下降，再加上e抗原滴度急剧减少。就在HBeAg血清转换，血清转化HBV-DNA水平低2个日志（P = 0.008），e抗原滴度310倍（P = 0.02），PSC突变> 25％（P <0.001），病毒进化的8.1倍（ P = 0.01）和9.3倍，较高的病毒多样性比基准值（p = 0.011）的2.9倍（P <0.001），非血清转化相比，病毒的多样性。血清转化系统进化树显示聚类单独的时间点和更长的分支长度比非血清转化（P = 0.01）。正选择检测在五个八血清转化，但没有在非血清转化（P = 0.026）。有显着的负相关关系，病毒多样性（RS = -0.60，P <0.001）和HBV-DNA或大三阳（RS = -0.58，P = 0.006）水平; PSC突变呈正相关（RS = 0.38，P = 0.009 ）。显着的正相关关系随着时间的推移，病毒多样性（RS = 0.65，P <0.001），而呈负相关（RS = -0.627，P <0.001），HBV-DNA和HBeAg水平（RS = -0.512，P = 0.015） 。

结论累积病​​毒进化变化，HBeAg血清转换之前提供洞察事件，可能会影响治疗。