DDW2012:Intrahepatic NK Cell Activation Is Dependent on Levels of HBsAg in Chron

StephenW

Intrahepatic NK Cell Activation Is Dependent on Levels of HBsAg in Chronic HBeAg-Negative Hepatitis B
3:15  | 741 |
Eric T. Tjwa, Roeland Zoutendijk, Gertine van Oord, Joanne Verheij, Paula J. Biesta, Andrea Woltman, Andre Boonstra, Harry L. Janssen                                         
Affiliation
Erasmus Medical Centre Rotterdam, Rotterdam, Netherlands

Abstract:
Background
Chronic hepatitis B (CHB) is the result of a failing immune response towards the virus. Blood natural killer (NK) cells serve as first-line defence against viral intruders, and these cells are functionally impaired in blood of CHB patients. Since little is known on liver NK cells in CHB, we aimed to investigate the activation status and function of intrahepatic NK cells in both HBeAg positive and negative CHB.
Methods Liver cells were isolated from biopsy specimens of 56 CHB patients. Liver leukocytes were stimulated for 24 h with IL-12/-18. Specimens were also processed for routine histopathological grading and staging according to Ishak scoring. NK cell frequencies, activation status (CD69) and function of NK cells were analysed by flow cytometry.
Results
Our cohort consisted of 21 HBeAg positive and 35 HBeAg negative CHB patients.  As expected, the levels of HBV-DNA and HBsAg were significantly higher in HBeAg positive disease (p<0.01). There was a moderate to strong correlation between HBV-DNA and HBsAg in both HBeAg positive and -negative disease. Frequency of liver NK cells and activated liver NK cells, as evidenced by CD69 positivity, was resp. 1.5 and 6-fold  higher than in blood (p<0.001). Upon stimulation, the frequency of CD69 positive liver NK cells increased less in liver than in blood (1.1 vs 4.5-fold) regardless of HBeAg status. In line with this, the frequency of IFN-γ expressing NK cells upon stimulation was 3 fold higher in blood than in liver (p<0.001). There was no difference between HBeAg positive and -negative CHB. Liver NK cell activation and IFN-γ expression was significantly correlated in HBeAg negative CHB (r=0.50, p<0.05). In this population, also HBsAg quantification correlated with liver NK cell activation (r=-0.60, p<0.05), but not with cytokine-expression or with levels of HBV-DNA. Liver histology (grading and staging) correlated neither with liver NK cell activation nor cytokine-expression in both HBeAg positive and -negative disease.
Conclusions
Our findings demonstrate that frequency and activation of NK cells differ strongly between the liver and blood. We further show that in chronic HBV patients, IFN-γ  production is
compromised despite a high NK cell activation status, which suggests exhaustion of specific functions of liver NK cells in CHB. We demonstrate for the first time that in HBeAg-negative CHB but not in HBeAg positive CHB, increasing HBsAg levels coincide with less liver NK cell activation, suggesting a role for HBsAg in inflammation but not in pathology. As a consequence, monitoring HBsAg levels during antiviral therapy may reflect modulation of the antiviral intrahepatic immune response.

Disclosure(s):
Harry L. Janssen - Consulting: Bristol-Myers Squibb Co. , Gilead Sciences Inc, Novartis Pharmaceuticals, Roche Pharma AG, Santaris, Medtronic Inc.  , Kirin, Abbott, DebioPharm S.A.; Grant/Research Support: Bristol-Myers Squibb Co. , Gilead Sciences Inc, Novartis Pharmaceuticals, Roche Pharma AG, Santaris, Medtronic Inc. , Anadys, Innogenetics
The following people have nothing to disclose: Eric T. Tjwa, Roeland Zoutendijk, Gertine van Oord, Joanne Verheij, Paula J. Biesta, Andrea Woltman, Andre Boonstra

StephenW

肝内NK细胞活性是依赖对HBeAg阴性慢性乙型肝炎表面抗原水平
3时15 | 741 |埃里克·T. Tjwa，Roeland Zoutendijk，Van Oord提供Gertine，陈家慧，Verheij葆研究Biesta，安德烈Woltman，安德烈Boonstra，哈里L.詹森

打印尔
Erasmus医学中心，鹿特丹，荷兰鹿特丹

摘要：

背景慢性乙型肝炎（CHB）是一个失败对病毒的免疫反应的结果。血液自然杀伤细胞（NK）作为抗病毒入侵的第一线防御，这些细胞功能受损慢性乙型肝炎患者的血液中。由于很少在CHB肝脏NK细胞，我们的目的是探讨在HBeAg阳性和阴性慢性乙型肝炎肝内NK细胞的活化状态和功能。
方法肝细胞中分离出56例慢性乙型肝炎患者的活检标本。肝脏白细胞刺激24小时与IL-12/-18。标本进行常规病理组织学分级和分期处理伊沙克得分。流式细胞仪分析NK细胞频率，激活状态（CD69分子）和NK细胞的功能。
结果我们的世代包括e抗原阳性和35 HBeAg阴性慢性乙型肝炎患者的21。正如所料，在HBeAg阳性患者（P <0.01），HBV-DNA和HBsAg水平明显高于。有一个很强的相关性，HBV-DNA和HBeAg阳性和阴性乙肝表面抗原的适度。频率，肝脏NK细胞和活化肝脏NK细胞表面CD69阳性证明，是RESP。 1.5和6倍，比血液中高（P <0.001）。刺激后的CD69阳性的肝脏NK细胞的频率增加肝比（1.1比4.5倍），血液中不论HBeAg状态。在这一行，频率γ-干扰素表达NK细胞刺激后高于肝脏，血液中的3倍（P <0.001）。 HBeAg阳性和阴性CHB之间有没有差异。 HBeAg阴性慢性乙型肝炎（R = 0.50，P <0.05），肝脏NK细胞活性和γ-干扰素的表达显着相关。在这一人群中，乙肝表面抗原定量与肝脏NK细胞活性（R = -0.60，P <0.05），但与细胞因子的表达，或HBV-DNA的水平。既不肝脏NK细胞的活化也不在HBeAg阳性和阴性细胞因子的表达与肝组织学相关（分级和分期）。
结论我们的研究结果表明，不同频率和NK细胞的活化肝脏和血液之间的强烈。我们进一步的研究表明，慢性乙肝患者中，γ-干扰素生产
尽管NK细胞的激活状态，这表明用尽慢性乙型肝炎肝脏NK细胞的特定功能损害。我们表明，在HBeAg阴性慢性乙型肝炎，但不是在HBeAg阳性慢性乙型肝炎的首次增加，HBsAg水平配合肝脏NK细胞的活化，提示乙肝表面抗原在炎症反应中的作用，但不是在病理。因此，监测抗病毒治疗期间HBsAg水平可能反映的抗病毒药物肝内免疫反应的调节。