EBV转化慢性乙型肝炎患者B淋巴母细胞系的建立

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(LCL)。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CpGDNA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LCL的形态特征,利用流式细胞术分析LCL细胞膜表面分子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。

Establishment of immortalized B lymphoblast cell line from patients with chronic hepatitis B

AIM: To establish immortalized B lymphoblast cell line (BLCL) from 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 incubated with EB virus (EBV) in the presence of CpG DNA motifs and cyclosporin A (CysA) for about 28 days. Morphological characteristic of the established immortalized BLCL was observed by microscope and the expression of CD19 and CD23 on cellular surface was determined by flow cytometry. RESULTS: BLCL was successfully established and could be cultured and propagated for a long time in vitro. CONCLUSION: EBV-immortalized BLCL provides a resource of target cells for further research on the cellular immunity of patients with CHB.