慢性乙型肝炎患者HBV核衣壳蛋白和包膜蛋白特异性B细胞的比

StephenW

J Hepatol. 2019 Jul 23. pii: S0168-8278(19)30424-6. doi: 10.1016/j.jhep.2019.07.015. [Epub ahead of print]
Comparative characterization of B cells specific for HBV nucleocapsid and envelope proteins in patients with chronic hepatitis B.
Le Bert N1, Salimzadeh L2, Gill US3, Dutertre CA4, Fachetti F5, Tan A1, Hung M6, Novikov N6, Lampertico P5, Fletcher SP6, Kennedy PTF3, Bertoletti A7.
Author information

1
    Emerging Infectious Diseases Program, Duke-NUS Medical School, Singapore, Singapore.
2
    Emerging Infectious Diseases Program, Duke-NUS Medical School, Singapore, Singapore; Department of Medicine, Yong Loo Lin School of Medicine, National University of Singapore, Singapore.
3
    Barts Liver Centre, Barts and The London School of Medicine & Dentistry, Queen Mary University of London, London, UK.
4
    Emerging Infectious Diseases Program, Duke-NUS Medical School, Singapore, Singapore; Singapore Immunology Network, Singapore Agency for Science, Technology & Research (A*STAR), Singapore, Singapore.
5
    Gastroenterology and Hepatology Division, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, University of Milan, Milan, Italy.
6
    Gilead Sciences, Department of Biology, Foster City, CA, USA.
7
    Emerging Infectious Diseases Program, Duke-NUS Medical School, Singapore, Singapore; Singapore Immunology Network, Singapore Agency for Science, Technology & Research (A*STAR), Singapore, Singapore. Electronic address: [email protected].

Abstract
BACKGROUND & AIMS:

Knowledge about the regulation ofanti-HBV humoral immunity during natural HBV infection is limited. We recently utilized dual fluorochrome-conjugated HBsAg to demonstrate, in patients with chronic HBV (CHB) infection, the functional impairment of their HBsAg-specific B cells. However, the features of their HBcAg-specific B cells are unknown. Here we developed a method to directly visualize, select and characterize HBcAg-specific B cells in parallel with HBsAg-specific B cells.
METHODS:

Fluorochrome-conjugated HBcAg reagents were synthetized and utilized to detect directly ex vivo HBcAg-specific B cells in 36 CHB patients. The frequency, phenotype, functional maturation and transcriptomic profile of HBcAg-specific B cells was studied by flow cytometry, in vitro maturation assays and Nanostring based detection of expression of immune genes, which we compared with HBsAg-specific B cells and total B cells.
RESULTS:

HBcAg-specific B cells are present at higher frequency than HBsAg-specific in CHB patients and, differently to HBsAg-specific B cells, they mature efficiently into antibody-secreting cells in vitro. Their phenotypic and transcriptome profiles show that HBcAg-specific B cells are preferentially IgG+ memory B cells.However, despite their phenotypic and functional differences, HBcAg- and HBsAg-specific B cells of CHB patients share a mRNA expression pattern that differs from global memory B cells and is characterized by high expression of genes indicative of cross-presentation and innate immune activity.
CONCLUSIONS:

During chronic HBV infection, a direct relation exists between serological detection of anti-HBs and anti-HBc antibodies and quantity and function of their respective specific B cells. However, the transcriptomic analysis performed in HBsAg- and HBcAg-specific B cells suggests additional roles of HBV-specific B cells beyond the production of antibodies.
LAY SUMMARY:

Protection of viral infection necessitates the production of antibodies that are generated by specialized cells of the immune system called B cells. During chronic HBV infection, antibodies against the internal part of the virus (core or HBcAg) are detectable while the antibodies directed against the virus envelope (surface or HBsAg) are not present. Here we developed a method that allows us to directly visualize ex vivo the B cells specific for these two viral components, highlighting their differences and similarities, and showing how two components of the same virus can differently impact on the function of antiviral B cells.

Copyright © 2019. Published by Elsevier B.V.
KEYWORDS:

Chronic hepatitis B (CHB); HBV-specific B cells; HBcAg; HBsAg; Hepatitis B virus (HBV)

PMID:
    31348999
DOI:
    10.1016/j.jhep.2019.07.015

StephenW

J Hepatol。 2019年7月23日.pii：S0168-8278（19）30424-6。 doi：10.1016 / j.jhep.2019.07.015。 [印刷前的电子版]
慢性乙型肝炎患者HBV核衣壳蛋白和包膜蛋白特异性B细胞的比较表征
Le Bert N1，Salimzadeh L2，Gill US3，Dutertre CA4，Fachetti F5，Tan A1，Hung M6，Novikov N6，Lampertico P5，Fletcher SP6，Kennedy PTF3，Bertoletti A7。
作者信息

1
    新加坡杜克新加坡国立大学医学院新出现的传染病项目。
2
    新加坡杜克新加坡国立大学医学院新兴传染病项目;新加坡国立大学Yong Loo Lin医学院医学系。
3
    Barts肝脏中心，Barts和伦敦医学和牙科学院，伦敦大学玛丽皇后学院，英国伦敦。
4
    新加坡杜克新加坡国立大学医学院新兴传染病项目;新加坡免疫学网络，新加坡科学技术与研究机构（A * STAR），新加坡，新加坡。
五
    意大利米兰米兰大学消化内科和肝病学系，基金会IRCCS Ca'Granda Ospedale Maggiore Policlinico。
6
    Gilead Sciences，美国加利福尼亚州福斯特市生物系。
7
    新加坡杜克新加坡国立大学医学院新兴传染病项目;新加坡免疫学网络，新加坡科学技术与研究机构（A * STAR），新加坡，新加坡。电子地址：[email protected]。

抽象
背景与目的：

关于自然HBV感染期间抗HBV体液免疫调节的知识是有限的。我们最近利用双荧光染料结合的HBsAg来证实慢性HBV（CHB）感染患者的HBsAg特异性B细胞功能受损。然而，他们的HBcAg特异性B细胞的特征是未知的。在这里，我们开发了一种方法，可以直接观察，选择和表征与HBsAg特异性B细胞平行的HBcAg特异性B细胞。
方法：

合成荧光染料缀合的HBcAg试剂并用于直接检测36名CHB患者体内HBcAg特异性B细胞。通过流式细胞术，体外成熟测定和基于纳米串的免疫基因表达检测，研究HBcAg特异性B细胞的频率，表型，功能成熟和转录组学特征，我们将其与HBsAg特异性B细胞和总B细胞进行比较。
结果：

HBcAg特异性B细胞在CHB患者中以高于HBsAg特异性的频率存在，并且与HBsAg特异性B细胞不同，它们在体外有效成熟为抗体分泌细胞。他们的表型和转录组谱显示HBcAg特异性B细胞优先是IgG +记忆B细胞。然而，尽管他们的表型和功能差异，CHB患者的HBcAg和HBsAg特异性B细胞共享mRNA表达模式不同于全球记忆B细胞的特征在于指示交叉呈递和先天免疫活性的基因的高表达。
结论：

在慢性HBV感染期间，抗HBs和抗HBc抗体的血清学检测与其各自特异性B细胞的数量和功能之间存在直接关系。然而，在HBsAg-和HBcAg特异性B细胞中进行的转录组学分析表明HBV特异性B细胞在抗体产生之外的其他作用。
LAY总结：

保护病毒感染需要产生由称为B细胞的免疫系统的特化细胞产生的抗体。在慢性HBV感染期间，可检测到针对病毒内部部分（核心或HBcAg）的抗体，而不存在针对病毒包膜（表面或HBsAg）的抗体。在这里，我们开发了一种方法，允许我们直接观察离体这两种病毒组分特异性的B细胞，突出它们的差异和相似性，并显示同一病毒的两种成分如何对抗病毒B细胞的功能产生不同的影响。

版权所有©2019。Elsevier B.V.
关键词：

慢性乙型肝炎（CHB）; HBV特异性B细胞;核心抗原;乙肝表面抗原;乙型肝炎病毒（HBV）

结论：
    31348999
DOI：
    10.1016 / j.jhep.2019.07.015