Yi X, Wen C H, Gu S Q, Guo L, Tang L B, Wang W B
Infectious Diseases Department, Nanfang Hospital of Southern Medical University, Guangzhou 510515, China.
Zhonghua Gan Zang Bing Za Zhi. 2021 Mar 20;29(3):240-245. doi: 10.3760/cma.j.cn501113-20190717-00253.
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. 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. Compared with untransformed peripheral blood B cells, BCL had high expression levels of CD138, CD38 and CD27, and the difference was statistically significant ( < 0.05), while the level of IL-6 production was decreased, and the difference was statistically significant ( < 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 ( < 0.01). 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.
建立爱泼斯坦-巴尔病毒转化的外周血B细胞系(BCL),并探讨其表型特征、分泌抗体和细胞因子的能力以及呈递乙型肝炎病毒(HBV)抗原肽的能力。从HBV感染患者中分离外周血单个核细胞(PBMC)。将爱泼斯坦-巴尔病毒上清液进行孵育以构建BCL。通过流式细胞术检测CD19、CD138、CD38、CD27的表达以及IFN-γ、IL-10、IL-6的产生水平。将负载HBV抗原肽的BCL与体外扩增的自体T细胞进行孵育。采用细胞内染色法检测T细胞产生的干扰素-γ水平。与未转化的外周血B细胞相比,BCL的CD138、CD38和CD27表达水平较高,差异具有统计学意义(<0.05),而IL-6的产生水平降低,差异具有统计学意义(<0.01)。负载HBV抗原肽的BCL显著增强了体外扩增的自体T细胞产生干扰素-γ的能力,差异具有统计学意义(<0.01)。BCL高表达CD138、CD38和CD27,但其产生IL-6的能力下降。BCL可提高HBV特异性T细胞对HBV抗原肽的免疫应答效率,可作为乙型肝炎免疫研究的新工具。