ELISPOT及FACS在慢性乙型肝炎病毒感染者细胞免疫功能检测中的应用  

Application of ELISPOT and FACS in cellular immune response in patients with chronic hepatitis B virus infection

作　　者：丛丽媛 吴庆军 李芸 杨海燕 谢锐姬 杨富强 

机构地区：[1]解放军第四五八医院肝病研究所免疫缺陷病研究与特殊实验中心,广州510600 [2]达科为生物技术有限公司,深圳518054

出　　处：《现代免疫学》2009年第5期409-412,共4页Current Immunology

摘　　要：为探讨HBV感染者特异性细胞免疫功能的差异,选取以流感多肽刺激健康人外周血单个核细胞(PBMC),采用ELISPOT法检测其PBMC中IFN-γ分泌细胞的数量。在以流感多肽为刺激物建立的流感质控细胞库的调控下,以HBV核心抗原为刺激物,检测表面抗体阳性的HBV感染者(A组)、e抗原阳性的慢性HBV携带者(B组)、e抗原阴性的慢性HBV携带者(C组)三类研究对象的IFN-γ分泌细胞数量。同时,采用FACS方法对A、B、C组分别进行T淋巴细胞亚群及调节性T细胞(Treg)检测。发现A与B、C组相比,斑点形成细胞数(SFC)/106PBMC明显减少(P=0.000及P=0.022),C组斑点形成细胞数显著多于B组(P=0.000)。FACS检测结果A组CD4+T细胞数量及CD4/CD8比值A组显著高于B、C组,而Treg数量B组显著高于A、C组。表明通过ELISPOT及FACS对乙肝病毒感染者外周血T细胞进行数量上和功能上的研究,能够深入了解体机体的免疫状态,从而为临床治疗及判定疾病的发展、转归提供理论依据。To investigate the cellular immune response in subjects with chronic hepatitis B virus （HBV） infection, the peripheral blood mononuclear cells （PBMCs） of the healthy donors were stimulated with influenza peptide to choose the suitable one used as the quality control. The quantity of IFN-γ producing cells of PBMCs in response to hepatitis B core antigen （HBeAg） in subjects with HBV infection was detected by enzyme linked immunosorbent spot（ELlSPOT）assay, and the quantity of T cell and regulatory T cell （Treg） was detected by flow cytometery（FCM）. It was demonstrated that the spot forming cells （SFC） per 106 PBMCs of HBV HBsAb positive subjects （group A） were significantly lower than those of the HBeAg positive HBV carriers （group B） and those with HBeAg negative HBV carriers （group C） （P=0. 000 and P=0. 022）. The SFC of group C was also significantly higher than that of group B. The percentage of CD4^＋ T cells and the ratio of CD4 and CD8 in group A were both significantly higher than those of group B and C, while the percentage of Treg in group B was significantly higher than group A and C. It is concluded that the study on the immune state of subjects with HBV infection through investigating the quantity and function of PBMC, can thereby provide information for clinical treatment and judgment of disease development.

关 键 词：乙型肝炎 细胞免疫 酶联免疫斑点 流式细胞术 

分 类 号：R392-33[医药卫生—免疫学]