慢性乙型肝炎病毒感染者外周血单个核细胞特异产生γ-干扰素的研究  

The specifical production of interferon-γ by peripheral blood mononuclear cells from patients with chronic hepatitis B virus infection

作　　者：周晖[1] 李琴[2] 吴长有[2] 江楚文[1] 陈学福[3] 

机构地区：[1]广东省医学科学院广东省人民医院检验科,广州市510080 [2]中山大学中山医学院免疫学教研室热带病防治研究教育部重点实验室,广州市510180 [3]广东省医学科学院广东省人民医院感染科,广州市510080

出　　处：《实用医学杂志》2009年第24期4223-4225,共3页The Journal of Practical Medicine

基　　金：广东省社会发展领域科技计划项目(编号:73070)

摘　　要：目的:研究慢性乙型肝炎病毒(HBV)感染者外周血单个核细胞(PBMC)经HBV表面抗原(HBsAg)刺激后产生γ-干扰素(IFN-γ)的免疫反应。方法:选择免疫清除期、免疫耐受期慢性乙肝患者和HBsAb阳性健康体检者各10例,Ficoll分离液法分离PBMC,用HBsAg刺激培养。利用ELISA方法检测细胞培养上清液中IFN-γ的含量;用酶联免疫斑点实验(ELISPot)检测PBMC中IFN-γ产生细胞的频率;用流式细胞术检测PBMC中IFN-γ^+的细胞亚群。结果:经HBsAg刺激培养后,免疫耐受组PBMC产生IFN-γ含量、IFN-γ产生细胞的频率明显低于正常对照组、免疫清除组(均p<0.05),免疫清除组则明显高于正常对照组(P<0.05)免疫清除组、正常对照组PBMC中IFN-γ^+的细胞亚群以CD4-CD8^+、CD4^-CD8^-细胞为主,且免疫清除组高于正常对照组;免疫耐受组仅CD4^-CD8^-表达IFN-γ,且低于另两组。结论:免疫清除期与免疫耐受期慢性乙肝患者PBMCs经HBsAg刺激后产生IFN-γ的免疫反应不同。Objective To study the intefferon-γ（IFN-γ） produced by peripheral blood mononuclear cells（PBMC） from patients with chronic hepatitis B virus infection in response to hepatitis B surface antigen （HBsAg）. Methods 10 patients in immune clearance phase （immune clearance group）, 10 patients in immunotolerant phase （immune tolerant group） and 10 healthy persons with positive HBsAb （control group） were analyzed. PBMC were prepared using Ficoll density gradient centrifugation and cultured in the presence or absence of HBsAg. The level of IFN-γ in culture supernatants, the frequency of IFN-γ-producing cells, and the subpopulations of IFN-γ-producing cells were detected by ELISA, ELISPOT and flow eytometry, respectively. Results The PBMC stimulated with HBsAg from immune tolerant group produced lower level of IFN-γ and frequnce of IFN-γ-producing cells than those from the other two groups （both P 〈 0.05）. Level of IFN-γ and frequnce of IFN-γ-producing cells of immune clearance group were higher than those of control group （P 〈 0.05）. The main subpopulations of IFN-γ-producing cells in immune clearance group and control group were CD4^- CD8^＋ T cell and CD4^- CD8^- cells. The percentages of IFN-γ-producing cell subsets in immune clearance group were all higher than those in control group. Only CD4^-CD8^- cells with lower percentage expressed IFN-γ in immune tolerant group, as compared with the other two groups. Conclusion The IFN-γ produced by PBMC stimulated with HBsAg from patients with chronic HBV infection in immune clearance phase is significant different from those in immunotolerant phase.

关 键 词：肝炎 乙型 慢性 外周血单个核细胞 Γ-干扰素 HBSAG 

分 类 号：R512.62[医药卫生—内科学] R512.91[医药卫生—临床医学]