酶联免疫斑点法检测慢性乙型肝炎患者HBcAg特异性分泌γ-干扰素细胞及其临床意义  

The detection of HBcAg-specific interferon γ secreting cell in chronic hepatitis B patient by enzyme linked immune spot assay and its clinical significance

作　　者：张宜俊[1] 夏书奇 曾水娣[2] 施理[1] 祖萍[1] 刘惠萍[1] 易学瑞[1] 田增遂 朱义鑫 

机构地区：[1]解放军第458医院全军肝病中心,广东广州510602 [2]解放军第458医院进修生,广东广州510602 [3]深圳市达科为生物技术有限公司,广东深圳518057

出　　处：《检验医学》2006年第1期8-11,共4页Laboratory Medicine

摘　　要：目的观察免疫应答不同期的慢性乙型肝炎（CHB）患者外周血中HBcAg特异性γ-干扰素（IFNγ）分泌细胞数，探讨其在评价患者体内非溶细胞机制清除乙型肝炎病毒（HBV）能力中的临床意义。方法采用酶联免疫斑点法（ELISPOT）检测83例免疫耐受期、49例免疫清除期患者和18名健康人HBcAg特异性IFNγ分泌细胞。结果健康人HBcAg特异性IFNγ分泌细胞数为11—19个／4×10^5个外周单个核细胞（PBMC），62．2％的免疫耐受期CHB患者低于正常值，44．9％的免疫清除期CHB患者高于正常值。HBcAg特异性IFNγ分泌细胞数与HBV DNA水平未发现有相关性，而HBcAg特异性IFNγ分泌细胞数与HBeAg S／N值在一定范围内有正相关趋势。13例免疫耐受期患者经乙型肝炎疫苗及草分枝杆菌制剂治疗3—6个月后，12例患者HBcAg特异性IFNγ分泌细胞数有不同程度提高。结论采用ELISPOT测定HBcAg特异性IFNγ分泌细胞数可从一个侧面反映大多数CHB患者免疫应答能力，并可以作为治疗效果的评价指标。Objective To investigate the frequency of HBcAg-specific interferon γ,（IFNγ） secreting cell of the peripheral blood mononuclear cell（PBMC） of patients with chronic hepatits B（CHB） at different immune response phases, as an indication of the immune status of patients through the noncytolytic mechanism to eliminate the hepatitis B virus in vivo. Methods The frequency of HBcAg-specific IFN3, secreting cell in 83 cases of the patients in immune tolerant phase, 49 cases of patients in the immune clearance phase and 18 healthy persons were analyzed by enzyme linked immune spot（ELISPOT） assays. Results The frequency of HBcAg-specific IFNγ secreting cell was about 11-19/4 × 10^5 PBMC in healthy persons. The frequency of the patients in the immune tolerant phase was 62.2% less than that of the healthy persons, and the frequency of the patients in the immune clearance phase was 44.9% more than that of the healthy persons. There was no relationship between the frequency of HBcAg-specific IFNγsecreting cell and HBV DNA copies in plasma. While the frequency of HBcAg-specific IFN-γ secreting cell was related with the value of HBeAg S/N in a certain range. In 12 of 13 immune tolerant patients, the frequency of HBcAg-specific IFNγ, secreting cell increased in certain extent after 3-6 months of immunotherapy including injection of reconabinant hepatitis B vaccine and preparation of Mycobacterium phlei. Conclusions The detection of HBcAg-specific IFNγ secreting cell by ELISPOT assays may reflect the immune status of patients with CHB in certain extent. It could be a useful marker for evaluating the effect of the clinical treatment.

关 键 词：乙型肝炎 酶联免疫斑点法 Γ-干扰素 

分 类 号：R446.61[医药卫生—诊断学]