慢性乙型肝炎病毒感染者不同阶段γ干扰素及其诱导蛋白10表达变化

Expression of interferon gamma and interferon gamma-inducible protein-lO in patients with different stages of chronic hepatitis B virus infection

目的观察慢性HBV感染者不同临床阶段7干扰素及其诱导蛋白10（IP-10）表达变化，探讨其对患者免疫活化和肝炎重症化的影响。方法2010年1月至2011年12月河北医科大学第三医院收治的慢性HBV感染免疫耐受期、免疫活动期（CHB）和CHB重型患者各15例，收集患者血清和肝组织标本。实时PCR检测肝组织γ干扰素、IP-10mRNA表达，免疫组织化学染色观察肝组织了干扰素、IP-10蛋白表达和分布，ELISA法测定血清-γ干扰素、IP=10水平。结果血清IP-10、干扰素γ水平在免疫耐受期[（142．09±47．64）、（2．47±0．60）ng／L]、免疫活动期[（415．27±145．52）、（6．98±1．12）ng／L]、CHB重型[（658．33±213．52）、（10．78±1．19）ng／L]患者中依次升高，以CHB重型患者升高最为显著（F值分别为43．48、256．98，均P〈0．05）；肝组织中7干扰素、IP-10mRNA和蛋白表达变化趋势与血清中的变化趋势平行，均以CHB重型患者最高，其后依次为免疫活动期、免疫耐受期患者，各组间比较差异均有统计学意义（F值分别为693．85、210．21、433．05、214．46，均P％0．05）。相关分析显示，免疫活动期和CHB重型患者血清IP-10水平均与肝组织炎症活动度分级（r=0．76，P％0．05）、血清γ干扰素水平（r=0．77，P〈0．05）呈显著正相关。结论IP10参与机体抗HBV感染免疫过程，是慢性HBV感染免疫耐受期向免疫活动期过渡的重要分子标志，且影响肝脏免疫损伤程度和重症化。

Objective To observe the expressions of interferon gamma （IFN-γ） and IFN-γ-inducible protein 10 （IP-10） at different stages of chronic hepatitis B virus （HBV） infection, and to investigate the relationship between IP-10 with hepatic inflammation activity and hepatitis aggravation. Methods Fifteen chronic hepatitis B （CHB） patients, 15 asymptomatic HBV carriers （AsC） and 15 chronic severe hepatitis B （CSHB） patients were enrolled in the study from January 2010 to December 2011 in the Third Hospital of Hebei Medical University. The liver samples were collected by percutaneous needle biopsy or from liver transplantation. The IFN-γ and IP-10 mRNA expressions were measured by quantitative real-time polymerase chain reaction （PCR）. Localization and hemi-quantitative analysis of IFN-γ and IP-10 proteins were performed by immunohistochemistry staining. Concentrations of serum IFN-γ and IP-10 were quantified by enzyme linked immunosorbent assay （ELISA）. HBV markers and liver function were also evaluated for each patient. Results Serum IFN-γ and IP-10 concentrations increased significantly in CHB [（415.27±145.52） ng/L and （6. 98±1.12） ng/L, respectively] and CSHB [（658.33±213. 52） ng/L and （10. 78± 1. 19） ng/L, respectively] patients compared with AsC [（142. 09 ± 47. 64） ng/L and （2. 47±90. 60） ng/L, respectively] patients, and were highest in CSHB patients （F=43.48, 256.98; both P〈0.05）. Meanwhile, intrahepatic IFN-γ and IP-10 mRNA and protein expressions paralleled with IFN-γand IP-10 concentrations in the serum, which was highest in CSHB patients, followed by CHB and AsC patients （F=693. 85, 210. 21, 433. 05, 214. 46; all PM0.05）. Furthermore, Spearman linear correlation analysis showed that serum IP-10 level was positively correlated with both hepatic inflammation activity and serum IFN-γ concentration in CHB and CSHB patients （r=0.76 and r=0.77, respectively; both P 〈 0. 05）. Conclusions IP-10, one important immunologic marker of regulating anti-HBV activation, indicates progression from immune tolerance phase to immune clearance phase. Furthermore, it may affect the degree of inflammation and hepatitis aggravation.