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中华实验和临床感染病杂志(电子版) ›› 2018, Vol. 12 ›› Issue (01) : 28 -34. doi: 10.3877/cma.j.issn.1674-1358.2018.01.006

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临床论著

十色流式检测HIV/AIDS患者外周血中T淋巴细胞亚群及活化状态
宋洋子1, 孔雅娴1, 郝禹1, 姜钰1, 汪笛2, 徐玲2, 种雪静2, 戴国瑞2, 赵红心2, 曾辉1,()   
  1. 1. 北京 100015,首都医科大学附属北京地坛医院传染病学研究所
    2. 北京 100015,首都医科大学附属北京地坛医院感染中心
  • 收稿日期:2017-02-27 出版日期:2018-02-15
  • 通信作者: 曾辉
  • 基金资助:
    北京市医院管理局"青苗"计划专项经费资助(No. QML20161803); 北京市科技新星计划(No. Z171100001117016)

Ten-color flow cytometry for measuring T cell subpopulation and its activation state in the peripheral blood of HIV/AIDS patients

Yangzi Song1, Yaxian Kong1, Yu Hao1, Yu Jiang1, Di Wang2, Ling Xu2, Xuejing Chong2, Guorui Dai2, Hongxin Zhao2, Hui Zeng1,()   

  1. 1. Institute of Infectious Diseases, Beijing Ditan Hospital, Capital Medical University, Beijing 100015, China
    2. Infectious Center, Beijing Ditan Hospital, Capital Medical University, Beijing 100015, China
  • Received:2017-02-27 Published:2018-02-15
  • Corresponding author: Hui Zeng
  • About author:
    Corresponding author: Zeng Hui, Email:
引用本文:

宋洋子, 孔雅娴, 郝禹, 姜钰, 汪笛, 徐玲, 种雪静, 戴国瑞, 赵红心, 曾辉. 十色流式检测HIV/AIDS患者外周血中T淋巴细胞亚群及活化状态[J/OL]. 中华实验和临床感染病杂志(电子版), 2018, 12(01): 28-34.

Yangzi Song, Yaxian Kong, Yu Hao, Yu Jiang, Di Wang, Ling Xu, Xuejing Chong, Guorui Dai, Hongxin Zhao, Hui Zeng. Ten-color flow cytometry for measuring T cell subpopulation and its activation state in the peripheral blood of HIV/AIDS patients[J/OL]. Chinese Journal of Experimental and Clinical Infectious Diseases(Electronic Edition), 2018, 12(01): 28-34.

目的

建立十色流式方案检测HIV/AIDS患者外周血中T淋巴细胞亚群及活化状态。

方法

按多色流式配色原则,初步确立抗人CD3、CD4、CD8、CD45RA、CD25、CCR7、CD28、CD38、HLA-DR抗体及7-AAD的配色方案。采用1例HIV/AIDS患者外周血PBMC标本进行最适电压调节、荧光补偿设定和FMO对照;设定条件后,检测5例HIV/AIDS患者的外周血样本。

结果

建立检测人外周血中T淋巴细胞亚群及活化状态的最优十色流式染色方案,采用此方案检测5例HIV/AIDS患者外周血标本,分析T淋巴细胞各亚群比例及活化程度,发现不同细胞亚群中活化分子HLA-DR、CD38、CD28的表达模式存在显著差异。

结论

十色流式方案可检测HIV/AIDS患者外周血中T淋巴细胞亚群及其活化状态操作简便,结果可靠。

Objective

To establish the ten-color flow cytometry for measuring T cell subpopulations and their activated state in the peripheral blood of HIV/AIDS patients.

Methods

The monoclonal antibodies targeting cell surface antigen CD3, CD4, CD8, CD45RA, CD25, CCR7, CD28, CD38 and HLA-DR to establish the ten-color panel were used. One peripheral blood PBMC sample from a HIV/AIDS patient was applied for adjustment of voltage, compensation and FMO control. After that peripheral blood samples, of 5 patients with HIV/AIDS were analyzed by ten-color flow cytometry.

Result

The ten-color flow cytometric staining method was established for the measurement of T cell subpopulations and activation state in human peripheral blood. This method was applied to detect peripheral blood samples from 5 patients with HIV/AIDS and was able to evaluate the immune status of these patients. And then the percentage of T lymphocyte subgroup and the activation level were detected, respectively. The expression patterns of HLA-DR, CD38 and CD28 in different subpopulations were with significant difference.

Conclusions

The ten-color flow cytometry was a simple and reliable method for the analysis of T cell subpopulations and their activation in HIV/AIDS patients.

图1 荧光通道最适电压
图2 CompBeads单色染色自动调节补偿
图3 细胞单色染色进行补偿微调
图4 HIV/AIDS患者外周血T细胞分析策略
表1 5例HIV/AIDS患者外周血T细胞中各细胞亚群活化的情况(±s
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