N-糖基化突变乙肝病毒表面主要亲水区主要免疫逃逸

StephenW

N-Glycosylation Mutations within Hepatitis B Virus Surface Major Hydrophilic Region Contribute Mostly to Immune Escape

    De-Min Yu a ,
    Xin-Hua Lia, f,
    Vannary Mom b,
    Zhong-Hua Lu d,
    Xiang-Wei Liao a,
    Yue Han a,
    Christian Pichoud b,
    Qi-Ming Gong a,
    Dong-hua Zhang a,
    Yan Zhang e,
    Paul Deny b,
    Fabien Zoulimb, c,
    Xin- XinZhang a, Corresponding author contact information, E-mail the corresponding author, E-mail the corresponding author

    a Department of Infectious Disease, Institute of Infectious and Respiratory Diseases, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, People’s Republic of China
    b INSERM, U1052, 151 cours Albert Thomas, 69424 Lyon cedex 03, France
    c Lyon University and Hospices Civils de Lyon, Lyon, France
    d Wu Xi Hospital of Infectious Diseases, People’s Republic of China
    e Ministry of Education Key Laboratory of Systems Biomedicine, Shanghai Center for Systems Biomedicine (SCSB), Shanghai Jiao Tong University, People’s Republic of China
    f The Third Affiliated Hospital, Sun Yat-sen University, Guangzhou, People’s Republic of China



Abstract
Background & aims

HBV immune escape represents a challenge to prevention, diagnosis, and treatment of hepatitis B. Here, we analyzed the molecular and clinical characteristics of HBV immune escape mutants in a Chinese cohort of chronically infected patients.
Methods

Two hundred sixteen patients with HBsAg and anti-HBs were studied, with one hundred eighty-two HBV carriers without anti-HBs as a control group. Recombinant HBsAg bearing the most frequent N-glycosylation mutations were expressed in CHO and Huh7 cells. After confirming N-glycosylation at the most frequent sites (129 and 131), together with inserted mutations, functional analysis were performed to study antigenicity and secretion capacity.
Results

One hundred twenty-three patients had the wild-type HBs gene sequence, 93 patients (43%) had mutants in the major hydrophilic region (MHR), and 47 of the 93 patients had additional N-glycosylation mutations, which were transmitted horizontally to at least 2 patients, one of whom was efficiently vaccinated. The frequency of N-glycosylation mutation in the case group was much higher than that of the control group (47/216 Vs 1/182).Compared with wild-type HBsAg, HBsAg mutants reacted weakly with Anti-HBs using a chemiluminescent microparticle enzyme immunoassay. Native gel analysis of secreted virion in supernatants of transfected Huh7 cells indicated that mutants had better virion enveloping and secretion capacity than wild-type HBV.
Conclusion

Our results suggest that specific HBsAg MHR N-glycosylation mutations are implicated in HBV immune escape in a high endemic area.

StephenW

背景与目的

HBV免疫逃逸的预防，诊断和治疗乙型肝炎这里是一个挑战，我们在中国队列慢性感染患者的HBV免疫逃逸突变的分子和临床特征进行了分析。
方法

两百年16例HBsAg和抗-HBs进行了研究，与182乙肝病毒携带者无抗-HBs作为对照组。轴承最常见的N-糖基化突变的重组HBsAg CHO和Huh7细胞中表达。在确认最频繁的网站（ 129和131 ） ，连同插入突变的N-糖基化后，进行功能分析，研究的抗原性和分泌能力。
结果

百23例HBs阳性野生型基因序列，有93例（43％）突变体的主要亲水区（ MHR ） ，并有额外的N-糖基化突变，这是水平传播的93例患者中47至少有2名患者，其中一人被有效地接种疫苗。 N-糖基化突变的情况下组的频率远高于对照组（47/ 216与1 / 182 ） 。与野生型相比，乙肝表面抗原，乙肝表面抗原突变体的反应弱，抗-HBs ，用化学发光微粒子酶免疫分析。母语凝胶电泳分析表明，突变体转染Huh7细胞培养上清中分泌病毒颗粒有更好的病毒粒子包围和分泌能力比野生型HBV 。
结论

我们的研究结果表明，特定的HBsAg MHR N-糖基化突变HBV免疫逃逸在高流行区有牵连的。