HBV膜蛋白突变影响献血者隐匿性乙肝病毒的抗原性和表型

肝胆速递

http://www.sciencedirect.com/science/article/pii/S0168827812003637
Influence of mutations in hepatitis B virus surface protein on viral antigenicity and phenotype in occult HBV strains from blood donorsHBV膜蛋白突变影响献血者隐匿性乙肝病毒的抗原性和表型

    Cheng-Hao Huang1, †,
    Quan Yuan1, 2, †,
    Pei-Jer Chen3,
    Ya-Li Zhang1,
    Chang-Rong Chen4,
    Qing-Bing Zheng1,
    Shiou-Hwei Yeh3,
    Hai Yu1, 2,
    Yu Xue1,
    Yi-Xin Chen1, 2,
    Ping-Guo Liu5,
    Sheng-Xiang Ge1, 2,
    Jun Zhang1, 2, Corresponding author contact information, E-mail the corresponding author,
    Ning-Shao Xia1, 2, Corresponding author contact information, E-mail the corresponding author

    1 National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Life Science, Xiamen University, Xiamen, Fujian Province, China
    2 School of Public Health, Xiamen University, Xiamen, Fujian Province, China
    3 National Taiwan University College of Medicine, National Taiwan University, Taipei, Taiwan
    4 Xiamen Blood Service, Xiamen, Fujian Province, China
    5 Zhongshan Hospital, Medical College of Xiamen University, Xiamen 361004, China

Background & Aims

This study aimed at investigating mutations in the hepatitis B surface protein (HBsAg) in occult hepatitis B virus (HBV) infection (OBI) and their influence on viral antigenicity and phenotype.
Methods

The characteristics of 61 carriers with OBI (OBI group), 153 HBsAg(+) carriers with serum HBsAg ⩽100 IU/ml (HBsAg-L group) and 54 carriers with serum HBsAg >100 IU/ml (HBsAg-H group) from 38,499 blood donors were investigated. Mutations in the major hydrophilic region (MHR) of the viral sequences were determined. Thirteen representative MHR mutations observed in OBI sequences were antigenically characterized with a panel of monoclonal antibodies (MAbs) and commercial HBsAg immunoassays and functionally characterized in HuH7 cells and hydrodynamically injected mice.
Results

Of 61 OBI sequences, 34 (55.7%) harbored MHR mutations, which was significantly higher than the frequency in either the HBsAg-L (34.0%, p = 0.003) or the HBsAg-H group (17.1%, p <0.001). Alterations in antigenicity induced by the 13 representative MHR mutations identified in the OBI group were assessed by reacting recombinant HBV mutants with 30 different MAbs targeting various epitopes. Four out of the 13 mutations (C124R, C124Y, K141E, and D144A) strongly decreased the analytical sensitivity of seven commercial HBsAg immunoassays, and 10 (G119R, C124Y, I126S, Q129R, S136P, C139R, T140I, K141E, D144A, and G145R) significantly impaired virion and/or S protein secretion in both HuH7 cells and mice.
Conclusions

MHR mutations alter antigenicity and impair virion secretion, both of which may contribute to HBsAg detection failure in individuals with OBI.

肝胆速递

从献血者乙肝病毒表面蛋白对病毒的抗原性和隐匿性HBV株表型突变的影响

    诚浩Huang1，†，
    泉媛1，2，†，
    裴哲Chen3，
    雅李Zhang1，
    常荣Chen4，
    清冰Zheng1，
    琇慧Yeh3，
    海宇1，2，
    于学1，
    易新Chen1，2，
    平果Liu5，
    盛祥GE1，2，
    2011年06月Zhang1，2，通讯作者的联系信息，E-mail的通讯作者，
    宁邵霞，2，通讯作者的联系信息，E-mail的通讯作者

    1美国国家传染病诊断试剂与疫苗开发的，生命科学学院，厦门大学，厦门，福建，中国
    2公共卫生学院，厦门大学，厦门，福建，中国
    3国立台湾大学医学院，国立台湾大学，台北，台湾
    4厦门市血液中心，厦门，福建，中国
    5医学院附属中山医院，厦门大学，厦门361004，中国

背景与目的

本研究旨在调查突变的B型肝炎表面蛋白（HBsAg）的隐匿性乙肝病毒（HBV）感染（OBI），其对病毒的抗原性和表型的影响。
方法

61个运营商的特点与欧倍德（OBI组），153乙型肝炎表面抗原（HBsAg）携带者血清HBsAg（+）⩽100 IU /毫升（乙型肝炎表面抗原（HBsAg）-L组）和54血清HBsAg携带者> 100 IU /毫升（乙型肝炎表面抗原（HBsAg）-H组） 38,499献血者进行了调查。突变的病毒序列的主要亲水区（MHR）进行了测定。抗原十三代表MHR欧倍德序列中观察到的突变特征与单克隆抗体（MAbs）的面板和商业的HBsAg的免疫测定和功能特征在于HuH7细胞和流体动力学注入小鼠。
结果

61 OBI序列，有34名（55.7％）窝藏MHR突变的频率显着高于乙肝表面抗原-L（34.0％，P = 0.003）或乙型肝炎表面抗原（HBsAg），H组（17.1％，P <0.001）。针对不同表位的有30种不同的单克隆抗体反应重组HBV变异株的抗原诱导的13个代表鉴定的OBI组中的MHR突变的改变进行了评估。四的13个突变（C124R，C124Y，K141E和D144A），大大减少了分析灵敏度的七家商业HBsAg的免疫，和10（G119R，C124Y，I126S，Q129R，S136P，C139R，T140I，K141E，D144A，G145R ）显着减值的病毒粒子和/或S蛋白的分泌在两个HuH7细胞和小鼠。
结论

MHR突变改变的抗原性和破坏病毒颗粒的分泌，这两者都可能有助于在个人与欧倍德对HBsAg检测故障。

肝胆速递

http://onlinelibrary.wiley.com/doi/10.1111/j.1365-2893.2012.01595.x/abstract
Mutations associated with occult hepatitis B virus infection result in decreased surface antigen expression in vitro

    C. M. Martin1,
    J. A. Welge2,
    S. D. Rouster1,
    M. T. Shata1,
    K. E. Sherman1,
    J. T. Blackard1

Article first published online: 4 JUN 2012
    1

    Division of Digestive Diseases
    2

    Departments of Psychiatry and Environmental Health, University of Cincinnati College of Medicine, Cincinnati, OH, USA

*Jason Blackard, PhD, Division of Digestive Diseases, University of Cincinnati College of Medicine, ML 0595, 231 Albert Sabin Way, Cincinnati, OH 45267, USA. E-mail: [email protected]

Summary.  Occult hepatitis B virus (HBV) infection is characterized by the absence of detectable hepatitis B surface antigen (HBsAg) in the serum, despite detectable HBV DNA. Investigations of the mechanisms underlying the development of occult HBV infection are lacking in the current literature, although viral mutations in the surface region, resulting in decreased HBsAg expression or secretion, represent one potential mechanism. Wild-type HBsAg expression vectors were constructed from genotype-matched chronic HBV sequences. Site-directed mutagenesis was then utilized to introduce three genotype A mutations – M103I, K122R and G145A – associated with occult HBV infection in vivo, alone and in combination, into the wild-type HBsAg vectors. Transfection of Huh7 and HepG2 cell lines was performed, and cell culture supernatants and cell lysates were collected over 7 days to assess the effects of these mutations on extracellular and intracellular HBsAg levels. The G145A mutation resulted in significantly decreased extracellular and intracellular HBsAg expression in vitro. The most pronounced reduction in HBsAg expression was observed when all three mutations were present. The mutations evaluated in vitro in the current study resulted in decreased HBsAg expression and potentially increased hepatic retention and/or decreased hepatic secretion of synthesized HBsAg, which could explain the lack of HBsAg detection that is characteristic of occult HBV infection in vivo.

肝胆速递

相关的突变隐匿性乙肝病毒感染所致，在降低表面抗原表达的影响

    C. M. Martin1，
    J. A. Welge2，
    S. D. Rouster1，
    M. T. Shata1，
    K. E. Sherman1，
    J. T. Blackard1

在网上公布：2012年6月4日
    1

    科消化系统疾病
    2

    ，辛辛那提大学医学院的大学，辛辛那提，俄亥俄州，美国精神病学和环境健康部门

*贾森Blackard，博士，科消化系统疾病，辛辛那提大学医学院，ML 0595，231伟业萨宾方式，辛辛那提，OH 45267，USA。电子邮件：jason.blackard uc.edu

总结。隐匿性乙型肝炎病毒（HBV）感染的特征是检测到B型肝炎表面抗原（HBsAg）的血清中的情况下，尽管检测到HBV DNA。缺乏机制的发展，隐匿性HBV感染的调查在目前的文献中，虽然病毒变异的表面区域，从而降低了HBsAg的表达和分泌，是一个潜在的机制。野生型HBsAg表达载体的构建基因型相匹配的慢性HBV序列。定点突变介绍三种基因型A突变 -  M103I，K122R和G145A  - 隐匿性HBV感染的体内，单独和组合，然后利用到野生型乙肝表面抗原载体。转染Huh7和HepG2细胞，并进行细胞培养上清液和细胞裂解液收集了超过7天，以评估这些突变的细胞外和细胞内的HBsAg水平的影响。 G145A突变导致的显着减少的细胞外和细胞内的HBsAg在体外表达。最显着的减少乙肝表面抗原的表达，观察，当所有三个突变。在目前的研究在体外评估的突变导致降低HBsAg的表达，并可能增加肝保留和/或减少肝脏合成的乙肝表面抗原分泌的，这可以解释缺乏的特点是隐匿性HBV感染体内的乙肝表面抗原检测。