EB病毒转化外周血B细胞系可提呈乙型肝炎病毒抗原肽

Hepatitis B virus antigen peptide presentation by Epstein-Barr virus-transformed peripheral blood B cells

目的建立EB病毒转化外周血B细胞系(BCL),探索其表型特点,分泌抗体、细胞因子的能力与提呈乙型肝炎病毒(HBV)抗原肽的能力。方法分离HBV感染者外周血单个核细胞,EB病毒上清液孵育后构建BCL,流式细胞术检测CD19、CD138,CD38、CD27表达及其产生γ干扰素、白细胞介素(IL)-10,IL-6的水平。BCL加载HBV抗原肽后与体外扩增的自体T细胞共孵育,采用胞内因子染色检测T细胞产生y干扰素水平。采用多个相关样本Friedman检验或两相关样本Wilcoxon符号秩和检验进行统计学分析。结果与未转化的外周血B细胞相比,BCL表达CD138、CD38、CD27水平升高,差异有统计学意义(P<0.05),而产生IL-6水平下降,差异有统计学意义(P<0.01)。加载HBV抗原肽的BCL可显著增强体外扩增的自体T细胞产生γ干扰素能力,差异有统计学意义(P<0.01)。结论BCL高表达CD138、CD38和CD27,但其产生IL-6能力下降;BCL可提高HBV特异性T细胞对HBV抗原肽的免疫应答效率,作为乙型肝炎免疫研究的新工具。

Objective To establish an Epstein-Barr virus-transformed peripheral blood B cell line(BCL),and explore its phenotypic characteristics,the ability to secrete antibodies and cytokines,and the ability to present hepatitis B virus(HBV)antigen peptide.Methods Peripheral blood mononuclear cells(PBMCs)were isolated from patients with HBV infection.Epstein-Barr virus supernatant was incubated to construct BCL.The expression of CD19,CD138,CD38,CD27 and the production levels of IFN-γ,IL-10,IL-6 were detected by flow cytometry.BCL loaded with HBV antigen peptide was incubated with in vitro-expanded autologous T cells.Intracellular staining was used to detect the level of interferon-gamma produced by T cells.Results Compared with untransfbrmed peripheral blood B cells,BCL had high expression levels of CD138,CD38 and CD27,and the difference was statistically significant(P<0.05),while the level of IL-6 production was decreased,and the difference was statistically significant(P<0.01).BCL loaded with HBV antigen peptide had significantly enhanced the production of interferon-gamma by in vitro-expanded autologous T cells,and the difference was statistically significant(P<0.01).Conclusion BCL highly expresses CD138,CD38 and CD27,but its ability to produce IL-6 decreases.BCL can improve the immune response efficiency of HBV-specific T cells to HBV antigen peptide,and serve as a new tool for hepatitis B immune research.