CD3brightCD56 + T细胞相关联，在慢性乙肝患者聚乙二醇化干扰素

StephenW

CD3brightCD56+ T cells associate with pegylated interferon-alpha treatment nonresponse in chronic hepatitis B patients

• 
  Chuang Guo
• , Xiaokun Shen
• , Binqing Fu
• , Yanyan Liu
• , Yongyan Chen
• , Fang Ni
• , Ying Ye
• , Rui Sun
• , Jiabin Li
• , Zhigang Tian
• & Haiming Wei
  

• Scientific Reports 6, Article number: 25567 (2016)
• doi:10.1038/srep25567
• Download Citation
• 
  
  • Hepatitis B
  • Immunosuppression
    

Received:10 November 2015Accepted:19 April 2016Published online:13 May 2016


AbstractChronic hepatitis B (CHB) infection is a serious and prevalent health concern worldwide, and the development of effective drugs and strategies to combat this disease is urgently needed. Currently, pegylated interferon-alpha (peg-IFNα) and nucleoside/nucleotide analogues (NA) are the most commonly prescribed treatments. However, sustained response rates in patients remain low, and the reasons are not well understood. Here, we observed that CHB patients preferentially harbored CD3brightCD56+ T cells, a newly identified CD56+ T cell population. Patients with this unique T cell population exhibited relatively poor responses to peg-IFNα treatment. CD3brightCD56+ T cells expressed remarkably high levels of the inhibitory molecule NKG2A as well as low levels of CD8. Even if patients were systematically treated with peg-IFNα, CD3brightCD56+ T cells remained in an inhibitory state throughout treatment and exhibited suppressed antiviral function. Furthermore, peg-IFNα treatment rapidly increased inhibitory TIM-3 expression on CD3brightCD56+ T cells, which negatively correlated with IFNγ production and might have led to their dysfunction. This study identified a novel CD3brightCD56+ T cell population preferentially shown in CHB patients, and indicated that the presence of CD3brightCD56+ T cells in CHB patients may be useful as a new indicator associated with poor therapeutic responses to peg-IFNα treatment.


IntroductionThe hepatitis B virus (HBV) infects more than 350 million people worldwide and is a major cause of chronic liver disease1. Both the innate and adaptive immune responses in the host regulate HBV infection2. In the innate immune response, hepatic natural killer (NK) cells exert their antiviral function against HBV infection by killing infected cells and producing high cytokine levels, which both promote the pathogenesis of viral hepatitis3. In the adaptive immune response, HBV-specific CD8+ T cells lyse infected hepatocytes and control viral infection; indeed, impaired CD8+ T cell activity is associated with the establishment of chronic HBV infection4. In addition, regulatory T cells are increased and have an immunosuppressive effect on HBV-specific T helper cells in chronic hepatitis B (CHB) patients5. The findings described above provide valuable information for understanding HBV pathogenesis and immune-evasion mechanisms. However, immune indexes that reflect the therapeutic efficacy of HBV treatments have not been so reliable, and other ways to evaluate therapeutic efficacy are needed.
Thus far, only three major clinical regimens to treat HBV are available: peg-IFNα, nucleoside/nucleotide analogues (NA), and the combination of peg-IFNα plus NA therapy6. Unlike HCV treatment that has yielded encouraging results, the effect of various therapies on HBV has been rather poor regardless of the treatment strategy. For instance, loss of hepatitis B e antigen (HBeAg)—a readout of reduced viral infectivity after treatment—occurs in only 30% of HBeAg-positive CHB patients treated with peg-IFNα, while the remaining 70% do not respond to treatment7. However, the underlying reason for this treatment resistance in HBV patients remains unknown.
A subset of the human T cell population expresses CD56, an NK cell surface marker. Generally, CD56+ T cells constitute approximately 10% of peripheral blood T cells and nearly 50% of liver T cells8,9. Upon stimulation, CD56+ T cells are activated, proliferate, and exhibit cytotoxicity in an MHC-unrestricted manner10,11. Notably, CD56+ T cells are a superior latent source of IFN-γ, which is considered to be a main mediator of antiviral responses12. As an abundant T cell subset in the liver, CD56+ T cells inhibit hepatic viral infection and replication, including HBV and HCV13,14. Moreover, CD56+ T cells are competent to treat a number of various infectious diseases15,16,17,18,19.
Despite this observed antiviral function, however, effector immune cells are always weaker in the context of HBV infection. We previously reported that TGFβ1 enrichment in HBV-persistent patients reduced NKG2D/2B4 expression on NK cells, leading to NK cell suppression20. In CHB patients, high NKG2A expression on NK cells decreased NK cell cytotoxicity21. Additionally, CHB patients reportedly harbor CD56+ T cells that display significantly increased inhibitory T cell immunoglobulin mucin-3 (Tim-3) expression over those from healthy controls, and this expression is further upregulated in patients with acute-on-chronic liver failure22. Tim-3 expression on CD56+ T cells also closely correlated with elevated serum ALT levels (a readout of liver injury) in CHB patients. Taken together, we speculate that CD56+ T cells may be in diminished antiviral status in CHB patients.
In order to understand the state of the immune system in CHB patients during HBV therapy, we evaluated new cases of untreated CHB patients who were systematically treated with peg-IFNα for 48 weeks. We identified that CHB patients could be classified into the following two different groups based on the intensity of CD3 expression on their CD56+ T cells: the CD3brightCD56+ T cell– and CD3dimCD56+ T cell–harboring CHB patient groups. Interestingly, a higher percentage of CHB patients (55/85, 64.7%) preferentially harbored the CD3brightCD56+ T cells than healthy controls (10/33, 30.3%). We further found that CD56+ T cells played an important role in the host response to peg-IFNα therapy and that the presence of peripheral CD3brightCD56+ T cells counted against host control of HBV and predicted poor therapeutic response. Indeed, CD3brightCD56+ T cells appeared to be both phenotypically and functionally inhibited. CD3brightCD56+ T cells rapidly upregulated Tim-3 expression during peg-IFNα treatment, which might explain the observed CD3brightCD56+ T cell dysfunction. Taken together, we provide a possible immunological explanation as to why a majority of CHB patients have a poor therapeutic response to peg-IFNα and present a new clinical outcome indicator that may serve as an auxiliary measurement of the efficacy of peg-IFNα treatment.

StephenW

CD3brightCD56 + T细胞相关联，在慢性乙肝患者聚乙二醇化干扰素治疗无应答

    郭家庄，肖昆沉Binqing福，刘岩岩，王永炎陈，倪方，樱野，孙锐，李佳宾，志刚田及卫海明

    科学报告6，商品编号：25567（2016）
    DOI：10.1038 / srep25567
    下载文献
        肝炎BImmunosuppression

收稿日期：
    2015年11月10日
公认：
    2016年4月19日
网络发布时间：
    2016年5月13日

抽象

慢性乙型肝炎（CHB）感染是一个严重的世界性和普遍的健康关注，以及有效的药物，并打击急需这种疾病的发展战略。目前，聚乙二醇化干扰素α（PEG-IFNα）和核苷/核苷酸类似物（NA）是最常用的处方治疗方法。然而，在患者持续应答率仍然很低，而原因尚不清楚。在这里，我们观察到慢性乙型肝炎患者优先包庇CD3brightCD56 + T细胞，一个新发现的CD56 + T细胞群。患者这种独特的T细胞群体表现出相对较差的反应PEG-IFNα治疗。 CD3brightCD56 + T细胞表达显着高水平的抑制分子NKG2A的和CD8的低水平。即使患者进行了系统用PEG-IFNα治疗，CD3brightCD56 + T细胞仍然在整个治疗过程中抑制状态，并表现出抑制抗病毒的功能。此外，PEG-IFNα治疗迅速上CD3brightCD56 + T细胞，这与IFNγ的产生负相关，并有可能导致其功能障碍增加抑制TIM-3的表达。本研究确定了CHB患者优先所示的新颖CD3brightCD56 + T细胞群，并表示CD3brightCD56 + T细胞在慢性乙型肝炎患者的存在可以是差的治疗应答相关于PEG-IFNα治疗的新的指标是有用的。
介绍

乙型肝炎病毒（HBV）感染全世界超过3.5亿人，是慢性肝病disease1的主要原因。无论是在宿主先天和适应性免疫反应调节HBV infection2。在先天免疫反应，肝自然杀伤（NK）细胞杀死感染的细胞和生产高细胞因子水平，这既促进病毒hepatitis3的发病发挥它们的抗乙肝病毒感染的抗病毒作用。在适应性免疫应答，HBV特异性CD8 + T细胞裂解感染的肝细胞，并控制病毒感染;的确，受损的CD8 + T细胞活性与建立慢性HBV infection4相关联。此外，调节性T细胞会增加，对慢性乙型肝炎（CHB）patients5 HBV特异性T辅助细胞免疫的效果。上述研究结果为了解乙肝发病机理和免疫逃逸机制提供了有价值的信息。然而，反映乙肝病毒治疗的疗效免疫指标都没有这么可靠，还需要其他的方法来评估治疗效果。

迄今为止，只有三个主要的临床治疗方案来治疗HBV是可用的：PEG-IFNα，核苷/核苷酸类似物（NA），和PEG-IFNα加NA therapy6的组合。不像取得了令人鼓舞的成果HCV治疗，各种疗法对乙肝病毒的效果已经比较差，无论治疗策略。例如，乙型肝炎e抗原（HBeAg）降低病毒感染性的-a读出丢失后治疗发生在仅30％的PEG-IFNα治疗HBeAg阳性CHB患者，而其余70％的人不向treatment7响应。然而，对于在HBV患者这种治疗性的根本原因仍然不明。

人T细胞群的一个子集表示CD56，NK细胞表面标记。一般情况下，CD56 + T细胞占外周血T细胞的约10％和近50％的肝牛逼cells8,9的。在刺激时，CD56 + T细胞被激活，增殖，以及在一个MHC-无限制manner10,11表现出细胞毒性。值得注意的是，CD56 + T细胞是IFN-γ，这被认为是抗病毒responses12的主要介体的优异的潜源。由于肝脏丰富的T细胞亚群，CD56 + T细胞抑制肝病毒的感染和复制，包括HBV和HCV13,14。此外，CD56 + T细胞有能力治疗许多各种感染diseases15,16,17,18,19的。

尽管此观察的抗病毒的功能，但是，效应的免疫细胞在HBV感染的情况下始终弱。我们以前报道，在乙肝患者持续TGFβ1富集的NK细胞减少NKG2D / 2B4表达，导致NK细胞suppression20。在慢性乙肝患者，NK细胞NKG2A高表达降低NK细胞cytotoxicity21。此外，慢性乙肝患者据说怀有CD56 + T细胞显示显著上升抑制T细胞免疫球蛋白粘蛋白3（TIM-3）对这些健康对照的表达，而这种表达在患者进一步上调急性上慢性肝failure22。对CD56 + T细胞Tim-3的表达也与血清ALT水平CHB患者（肝脏损伤的读数）相关。综上所述，我们推测，CD56 + T细胞可能是在慢性乙肝患者抗病毒减弱的状态。

为了了解HBV治疗期间慢性乙型肝炎患者免疫系统的状态，我们评估了谁进行了系统用PEG-IFNα治疗48周未治疗慢性乙型肝炎患者的新发病例。我们确定了慢性乙型肝炎患者可分为基于CD3表达对他们的CD56 + T细胞的力度以下两种不同的群体：CD3brightCD56 + T细胞和CD3dimCD56 + T细胞窝藏CHB患者群。有趣的是，慢性乙肝患者（55/85，64.7％）的比例更高优先窝藏CD3brightCD56 + T细胞比正常对照组（10/33，30.3％）。我们还发现，CD56 + T细胞在宿主响应于钉IFNα疗法和计数抗HBV的主机控制外围CD3brightCD56 + T细胞的存在发挥了重要作用，并预测差的治疗反应。事实上，CD3brightCD56 + T细胞似乎是两个表型上和功能上被抑制。 PEG-IFNα治疗期间CD3brightCD56 + T细胞迅速上调Tim-3的表达，这或许可以解释观测到的CD3brightCD56 + T细胞的功能障碍。总之，我们提供了一个可能的免疫学解释为什么大多数CHB患者有不良的治疗反应挂-IFNα，并提出了一种新的临床结果的指标，可作为PEG-IFNα治疗的疗效的辅助测量。