慢性乙型肝炎患者树突状细胞培养与功能检测方法  

The Method of Culturing and Testing the Ability of Dendritic Cells from Chronic Hepatitis B Patients

作　　者：王麟[1] 赵中夫[2] 卢俊敏[1] 

机构地区：[1]山西医科大学,030001 [2]长治医学院肝病研究所

出　　处：《长治医学院学报》2008年第3期172-174,共3页Journal of Changzhi Medical College

基　　金：山西省自然科学基金(2008011073-4)

摘　　要：目的:建立从人外周血分离培养树突状细胞(dendriticcell,DC)及检测其表型与诱导淋巴细胞增殖能力的方法。方法:从未接受抗病毒治疗的CHB患者外周血中分离单个核细胞,在含GM-CSF、IL-4、TNF-α的完全培养基中诱导培养为DC,流式细胞仪检测细胞表型,混合淋巴细胞反应检测DC刺激淋巴细胞增殖的能力。结果:CHB患者外周血单核细胞,在体外可诱导出具有典型形态及生物学活性的DC。患者DC的HLA-DR、CD80、CD86的表达率分别为(44.77±8.127)%、(36.65±6.867)%、(32.16±6.340)%,CHB患者DC刺激淋巴细胞增殖的SI为1.39±0.375。与正常人DC相比明显降低(P<0.01)。结论:CHB患者单个核细胞能被诱导培养为DC;通过检测DC表型和刺激淋巴细胞增殖能力可以了解CHB患者抗HBV免疫功能。Objective：To establish the method of culturing DC from human peripheral blood and testing the phenotype and the capacity of stimulating leukocyte.Methods：Selecting CHB without receiving antiviral treatment,PBMCs were separated from peripheral blood,and then cultured in vitro with medium containing GM-CSF IL-4 TNF-αinto DC.The phenotype of DC was identified by flow cytometry.The abilities to stimulate T cell proliferate were detected by mixed leukocytes reaction（MLR）.Results：PBMC from CHB patients which were cultured in vitro could be induced into typical DC.The expression of costimulating molecules CD80、CD86、HLA-DR in the surface of DCs from CHB patients before antiviral treatment were（32.16±6.340）%、（36.65±6.867）%、（44.77±8.127）%.The stimulation index（SI）that DC stimulate lymphocytes to proliferate was 1.39±0.375.Conclusion：The PBMC from CHB patients could be induced into DC.Testing the phenotype and the capacity of stimulating leukocyte of DC from CHB can be known the ability of antiviral immunity.

关 键 词：树突状细胞 慢性乙型肝炎 免疫状态 

分 类 号：R446.6[医药卫生—诊断学]