摘要
目的体外建立慢性乙型肝炎患者永生化的B淋巴母细胞系(LCLs)。方法用EB病毒感染自慢性乙型肝炎患者外周血中分离的单个核细胞(PBMC),加入CpG DNA免疫调节基序以诱导B淋巴细胞增殖,环胞菌素A(CysA)抑制T淋巴细胞的活性。光学显微镜下观察LCLs的形态特征,利用流式细胞术分析LCLs膜表面分子CD19和CD23的表达水平。结果46例患者PBMC经EBV感染4周后,42例转化成永生化B淋巴母细胞系,成功率为91.3%。转化后的B淋巴母细胞体积增大积聚成团,可进一步分裂增殖并长期传代培养。结论CpG免疫调节基序联合EBV感染人PBMC,提高转化效率,转化后的LCL保持了成熟B淋巴细胞的生物学特性,可作为体外研究HBV特异性免疫应答的刺激细胞和靶细胞。
Objective To establish the Epstein-Barr virus （EBV） transformed B lymphoblastoid cell lines （LCLs） in patients with chronic hepatitis B （CHB） in vitro. Methods The peripheral blood mononuclear cells （PBMC） were isolated from the patients with CHB by routine method, and then incubated with EBV in the presence of CpG DNA motifs and cyclosparin A （CysA） for about 28 d. Morphological characteristics of immortalized B-lymphocytes were observed with microscope. Expressions of surface molecules CD19 and CD23 in LCLs were determined by flow cytometry. Results LCLs were successfiAly established and could be cultured and propagated for a long time in vitro. Conclusion EBV-immortalized LCLs provide target cells for further research on the cellular immune function of hepatitis B patients.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2007年第4期462-465,共4页
Immunological Journal
基金
全军医学科学技术研究"十五"计划重点课题(01Z046)
