目的检测慢性乙型肝炎患者外周血单个核细胞(PBMC)-α干扰素(IFN-α)的诱导表达及白细胞介素12(IL-12)表达,评价慢性乙肝患者PBMC的细胞免疫功能。方法poly IC体外刺激正常对照组和慢性乙肝病人PBMC,取培养上清利用病毒保护试验测定干扰素治疗前后病人PBMC表达的干扰素抗病毒生物学活性。同时在IFN-α治疗前后,用RT-PCR检测慢性乙肝患者不同干扰素应答组病人PBMC IL-12表达的变化。结果干扰素治疗前应答组病人(n=4)和无应答组病人(n=12)PBMC分泌的干扰素生物学活性显著低于正常对照组(n=10)(P〈0.01)。应答组病人PBMC分泌的干扰素活性随治疗时间延长而增加,1个月时已显著高于无应答组病人(P〈0.01);无应答组病人PBMC分泌的干扰素活性始终处于低水平。治疗前,应答组和无应答组病人PBMC的IL-12 mRNA水平都显著高于正常对照组(P〈0.01);治疗后,应答组病人IL-12水平随治疗时间延长而降低,1个月时已显著低于无应答组病人(P〈0.05),无应答组病人IL-12水平始终处于较高水平。结论慢性乙型肝炎病人的细胞免疫功能受损,PBMC不能正常表达IFN-α,但应答组病人经干扰素治疗后IFN-α表达能力逐渐恢复。慢性乙肝患者PBMC IL-12表达与肝脏炎症活动有关。慢性乙肝患者PBMC IFN-α活性检测结果也许可以作为干扰素治疗效果的预测指标。
Objective To detect the expression of interferon-α(IFN-α) and interleukin 12 (IL-12) in peripheral blood mononuclear cells(PBMCs) of patients with chronic hepatitis B and analyze cellular immune status of the patients with chronic hepatitis B virus (HBV) infection. Methods The PBMCs of patients with chronic HBV infection were cultured, then the antiviral biological activity of PBMCs stimulated with high concentration of PolyIC in the culture supernatants was detected by viral protection assay. IL-12 mRNA in PBMCs from the patients with chronic hepatitis B was measured by reverse transcription polymerase chain reaction(RT-PCR). Results The obtained data indicated that lower titer of IFN-αfrom PolyIC-induced PBMCs existed in patients infected by chronic HBV. Enhanced IFN-αactivity from PBMCs stimulated by PolyIC was found in the responder( n = 4) who were treated with IFN-α2b. However, continuously lower activity of IFN-αfrom PBMCs presented in non-responder ( n = 12) during IFN-α2b therapy. The expressions of IL-12 mRNA in PBMCs from both responder and non-responder were significantly higher than those of normal control group ( n = 10) before treatment with IFN-α2b ( P 〈 0.01 ). However, after treating with IFN-α2b,the level of IL-12 mRNA of responder decreased but the level of IL-12 mRNA of non-responder kept high. Conclusions The cellular immune function of the patients with chronic HBV infection is impaired. The expression of IFN-αgene in non-responder of the patients with chronic HBV infection was inhibited. The level of IL-12 was consistent with the activity of liver inflammation. The IFN-α activity of PBMCs may be a marker for the prognosis of disease in the chronic hepatitis B patients after IFN-αtherapy.