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 untransformed 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.
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γ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。.
Keywords: Antigen-presenting cells; Epstein-Barr virus; Hepatitis B virus; Peripheral B cell lines.