单　奔，孔歌，李　彦，王　霞，傅涓涓，李　丽，潘修成

·乙型肝炎·

HBeAg阳性慢性乙型肝炎患者血清Mig、RANTES和IL-9水平与HBV前C/BCP区变异的关系及其对聚乙二醇α-干扰素治疗应答的影响*

单奔，孔歌，李彦，王霞，傅涓涓，李丽，潘修成

目的研究HBeAg阳性CHB患者血清干扰素-γ诱导的单核因子（Mig）、调节活化正常T细胞表达和分泌的趋化因子（RANTES）和IL-9水平与HBV前C区和BCP区变异之间关系及其对聚乙二醇α-干扰素治疗应答的影响。方法纳入43例接受聚乙二醇α-干扰素治疗的HBeAg阳性CHB患者，采用PCR技术扩增治疗前患者血清HBV前C区和BCP区基因片段，并测序分析。采用微量样本多指标流式蛋白定量（CBA）技术检测血清Mig、RANTES和IL-9水平。结果在43例HBeAg阳性CHB患者中，检测到HBV前C区和（或）BCP区野生型（WT）24例（55.8%）和突变型（MT）19例（44.2%）；野生组血清RANTES和IL-9基线水平分别为（3274.24±814.79）pg/mL和（9.40±0.89）pg/mL，突变组则为（2742.40±764.24）pg/mL和（10.78±2.73）pg/mL，两组间差异均具有统计学意义（P<0.05）；在治疗48 w时，突变组患者获得CR者9例（47.4%），野生组患者4例（16.7%），两组间差异有统计学意义（P<0.05）；突变组和野生组对聚乙二醇α-干扰素治疗实现完全应答的CHB患者IL-9、Mig和RANTES基线水平均无显著性差异；突变组中治疗24 w时HBsAg下降幅度>1 log10的CHB患者血清Mig基线水平为（138.17±96.57）pg/m L，而HBsAg下降<1 log10的CHB患者为（89.74±78.25）pg/mL，两组间差异有统计学意义（P<0.05）；聚乙二醇α-干扰素治疗过程中，突变组血清IL-9基线水平为（10.78±2.73）pg/mL，治疗12 w和24 w时分别为（8.83±1.94）pg/mL和（8.91±1.97）pg/mL，均明显低于基线水平（P<0.05），而野生组中未发现有类似的变化。结论HBV前C或（和）BCP区变异与IL-9水平升高密切相关，但是IL-9水平与聚乙二醇α-干扰素抗病毒的疗效无关。Mig基线高水平可能有利于突变组CHB患者在聚乙二醇α-干扰素治疗过程中实现HBsAg的清除。

慢性乙型肝炎；病毒变异；聚乙二醇α-干扰素；白细胞介素-9；干扰素-γ诱导单核因子

【Abstract】Objective To study the correlation of serum monokine induced by IFN-γ(Mig),regulated upon activation normal T-cell expressed and secreted factors(RANTES)and IL-9 levels to HBV pre-C/BCP mutation and their influences on the response to Peg-IFN-αtherapy in patients with HBeAg-positive hepatitis B. Methods A total of 43 HBeAg positive patients with CHB who received Peg-IFN-αtherapy were enrolled in this study.Serum HBV DNA was extracted from peripheral blood,and polymerase chain reaction(PCR)was performed to sequence the Pre-C/BCP gene fragments.Serum levels of Mig,RANTES and IL-9 were detected by using cytometric bead array(CBA).Resu lts The mutation(MT)of HBV Pre-C and/or BCP was positive in 19 (44.2%)patients,and the wild type was 24(55.8%)out of the 43 HBeAg-positive hepatitis B patients;the serum levels of RANTES and IL-9 were(3274.24±814.79)pg/mL and(9.40±0.89)pg/mL,respectively,in patients withHBV pre-C/BCP mutation,and were(2742.40±764.24)pg/mL and(10.78±2.73)pg/mL,respectively,in patients without HBV pre-C/BCP mutation,and the difference was statistically significant(P<0.05);nine(47.4%)out of patients with and 4(16.7%)out of those without viral mutation obtained complete response at the end of 48 week regimen(P<0.05).The baseline level of serum Mig in patients with serum HBsAg decline at week 24 of Peg-IFN-αtreatment was(138.17±96.57)pg/mL,and the baseline level in those without HBsAg decline was (89.74±78.25)pg/mL(P<0.05);In patients with HBV pre-C/BCP mutation,the baseline level of IL-9 was(10.78± 2.73)pg/mL,the levels at week 12 and 24 during the treatment were(8.83±1.94）pg/mL and(8.91±1.97)pg/m L, both significantly lower than the baseline(P<0.05).Conclusion Serum level of IL-9 is closely associated with HBV Pre-C/BCP mutation,but not with anti-HBV efficacy of Peg-IFN-αtreatment in HbeAg-positive patients with CHB.Higher baseline level of Mig could contribute to HBsAg decline after PegIFN-αtherapy in CHB patients with HBV pre-C/BCP mutation.

【Key words】Hepatitis B;Mutation;PEG-IFN-α;Interleukin 9;Monokine induced by IFN-γ

α-干扰素是HBeAg阳性慢性乙型肝炎（CHB）患者的一线抗病毒治疗药物，约有30%患者在治疗结束后获得持久性应答[1]。已证实HBeAg阳性CHB患者体内有HBV前C区1896位或/和基本核心启动子（Basal core promoter,BCP）区1762或/和1764位基因变异，可降低或终止HBeAg的产生，进而影响对α-干扰素的应答反应[2]。HBV感染过程与宿主免疫状态密切相关，而细胞（趋化）因子网络在慢性乙型肝炎发病过程中发挥着重要作用，并影响机体对α-干扰素抗HBV的应答反应[3]。然而，至今尚不完全明确HBV前C/BCP区变异与细胞因子水平之间的确切关系及其对干扰素抗HBV疗效的影响。……