北京协和医学院比较医学中心,中国医学科学院医学实验动物研究所,国家卫生健康委员会人类疾病比较医学重点实验室, 国家中医药管理局人类疾病动物模型三级实验室,新发再发传染病动物模型研究北京市重点实验室,北京 100021
R-33
LI Guocui
Comparative Medicine Center, Peking Union College (PUMC)&Institute of Laboratory Animal Science, Chinese Academy of Medical Sciences (CAMS); NHC Key Laboratory of Human Disease Comparative Medicine; Key Laboratory of Human Diseases Animal Models, State Administration of Traditional Chinese Medicine; Beijing Key Laboratory for Animal Models of Emerging and Remerging Infectious Diseases, Beijing 100021, ChinaLU Jiahan
Comparative Medicine Center, Peking Union College (PUMC)&Institute of Laboratory Animal Science, Chinese Academy of Medical Sciences (CAMS); NHC Key Laboratory of Human Disease Comparative Medicine; Key Laboratory of Human Diseases Animal Models, State Administration of Traditional Chinese Medicine; Beijing Key Laboratory for Animal Models of Emerging and Remerging Infectious Diseases, Beijing 100021, ChinaLU Qiuhan
Comparative Medicine Center, Peking Union College (PUMC)&Institute of Laboratory Animal Science, Chinese Academy of Medical Sciences (CAMS); NHC Key Laboratory of Human Disease Comparative Medicine; Key Laboratory of Human Diseases Animal Models, State Administration of Traditional Chinese Medicine; Beijing Key Laboratory for Animal Models of Emerging and Remerging Infectious Diseases, Beijing 100021, ChinaYANG Chenbo
Comparative Medicine Center, Peking Union College (PUMC)&Institute of Laboratory Animal Science, Chinese Academy of Medical Sciences (CAMS); NHC Key Laboratory of Human Disease Comparative Medicine; Key Laboratory of Human Diseases Animal Models, State Administration of Traditional Chinese Medicine; Beijing Key Laboratory for Animal Models of Emerging and Remerging Infectious Diseases, Beijing 100021, ChinaCONG Zhe
Comparative Medicine Center, Peking Union College (PUMC)&Institute of Laboratory Animal Science, Chinese Academy of Medical Sciences (CAMS); NHC Key Laboratory of Human Disease Comparative Medicine; Key Laboratory of Human Diseases Animal Models, State Administration of Traditional Chinese Medicine; Beijing Key Laboratory for Animal Models of Emerging and Remerging Infectious Diseases, Beijing 100021, ChinaWEI Qiang
Comparative Medicine Center, Peking Union College (PUMC)&Institute of Laboratory Animal Science, Chinese Academy of Medical Sciences (CAMS); NHC Key Laboratory of Human Disease Comparative Medicine; Key Laboratory of Human Diseases Animal Models, State Administration of Traditional Chinese Medicine; Beijing Key Laboratory for Animal Models of Emerging and Remerging Infectious Diseases, Beijing 100021, ChinaComparative Medicine Center, Peking Union College (PUMC)&Institute of Laboratory Animal Science, Chinese Academy of Medical Sciences (CAMS); NHC Key Laboratory of Human Disease Comparative Medicine; Key Laboratory of Human Diseases Animal Models, State Administration of Traditional Chinese Medicine; Beijing Key Laboratory for Animal Models of Emerging and Remerging Infectious Diseases, Beijing 100021, China
目的 比较不同诱导刺激剂对 T 细胞活化标志物 CD69、CD38、HLA-DR 和核内增殖标记 Ki67 的表达量的影响,为研究 T 细胞激活功能和作用研究提供依据。 方法 采集健康恒河猴外周血,分离 PBMC,分别加入常规剂量的 PMA+ Ionomycin、α-CD2 / α-CD3 / α-CD28 和 PHA-P+IL-2,在不同的时间点检测 CD4+T 细胞和 CD8+T 细胞表面标志物 CD69、CD38、HLA-DR 和核内增殖标记 Ki67 表达情况。 结果 流式细胞术结果表明,在刺激剂作用 72 h 时,与其它两种刺激剂相比,α-CD2 / α-CD3 / α-CD28 刺激后的 CD4+T 细胞和 CD8+ T 细胞上的 CD69 和 Ki67 表达量更高,HLA-DR+CD4+T 细胞和 HLA-DR+CD8+T 细胞的比例与另外两组组间比对无差异,同时 CD38+ CD4+T 细胞和 CD38+CD8+T 细胞的比例在 α-CD2 / α-CD3 / α-CD28 与 PHA-P+IL-2 组间比较无统计学差异(P>0. 05);在 PMA +Ionomycin 刺激下,检测到上述四种分子的表达量都很低。 结论 CD2 / α-CD3 / α-CD28 活化 T 细胞效果最佳, PHA-P+IL-2 次之,PMA+Ionomycin 作用弱。 在选择 T 细胞活化作用程度方面,可进行针对性选择。
Objective To investigate the effect of stimulants on the expression of T cell activation markers (CD69, CD38, HLA-DR) and a nuclear proliferation marker ( Ki67), and provide the basis for the study of T cell function after activation. Methods Peripheral blood was collected from healthy rhesus monkeys, then peripheral blood mononuclear cells ( PBMCs ) were separated and regular doses of PMA + Ionomycin, α-CD2 / α-CD3 / α-CD28, and phytohemagglutinin-P (PHA-P)+interleukin-2 ( IL-2) were added. Expression of CD4+ T cells and CD8+T cell surface markers, CD69, CD38, HLA-DR, and Ki67, were detected at different time points. Results Flow cytometry showed that when the stimulant, α-CD2 / α-CD3 / α-CD28, acted for 72 hours, expression of CD69 and Ki67 on CD4+T cells and CD8+T cells was higher compared with the other two stimulants, while the percentage of HLA-DR+ CD4+T cells and HLA-DR+ CD8+T cells and CD38+CD8+T cells was not different. Meanwhile, the percentage of CD38+CD4+T cells was not different between α-CD2 / α-CD3 / α-CD28 and PHA-P + IL-2 groups ( P> 0. 05). Expression of all four molecules was very low under PMA+Ionomycin stimulation. Conclusions CD2 / α-CD3 / α-CD28 had the best effect in activating T cells, followed by PHA-P+IL-2, and then PMA+Ionomycin. This provides the experimental basis for selecting appropriate stimulants on the basis of different experimental purposes.
李国萃,陆佳涵,卢秋翰,杨晨波,丛 喆,魏 强.诱导刺激猴外周血单核淋巴细胞亚群活化增殖研究[J].中国比较医学杂志,2021,31(5):1~6.
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