基础医学与临床 ›› 2009, Vol. 29 ›› Issue (1): 24-28.

• 研究论文 • 上一篇    下一篇

一株健康人γδT细胞克隆的建立及鉴定

何小鹃 康宁 陈慧 崔莲仙 何维   

  1. 中国医学科学院基础医学研究所 北京协和医学院基础学院 中国医学科学院基础医学研究所 北京协和医学院基础学院 中国医学科学院基础医学研究所 北京协和医学院基础学院 中国医学科学院基础医学研究所 北京协和医学院基础学院 中国医学科学院基础医学研究所 北京协和医学院基础学院
  • 收稿日期:2008-07-14 修回日期:2008-09-04 出版日期:2009-01-25 发布日期:2009-01-25
  • 通讯作者: 何维

Establishment and characterization of healthy donor's γδT cell clones

Xiao-juan HE, Ning KANG, Hui CHEN, Lian-xian CUI, Wei HE   

  1. Institute of Basic Medical Science & School of Basic Medicine, CAMS & PUMC Institute of Basic Medical Science & School of Basic Medicine, CAMS & PUMC Institute of Basic Medical Science & School of Basic Medicine, CAMS & PUMC Institute of Basic Medical Science & School of Basic Medicine, CAMS & PUMC Institute of Basic Medical Science & School of Basic Medicine, CAMS & PUMC
  • Received:2008-07-14 Revised:2008-09-04 Online:2009-01-25 Published:2009-01-25
  • Contact: Wei HE

摘要: 目的 建立健康人γδT细胞克隆并进行鉴定。方法 以60Co照射的同种异体PBMC作为饲养细胞,以有限稀释法对经过阳性分选的γδT细胞进行克隆化。用流式细胞术及分子生物学检测鉴定所获克隆,以MTT掺入法对所获克隆进行增殖实验检测。结果 获得一株γδT细胞单克隆,为Vγ9Vδ2亚型,CD4分子和CD8分子表达均为阴性。进一步的研究发现表位肽EP6不仅能够特异性结合γδT细胞单克隆,而且能够促进其增殖。结论 成功建立了健康人外周血γδT细胞克隆,为深入研究γδT细胞抗原识别机制、亚群及功能奠定了坚实的基础。

关键词: γδT细胞, 克隆, 有限稀释

Abstract: Objective Establishment and characterization of healthy donor's γδT cell clones. Methods γδT cells were cloned by limiting dilution after positive sorting, with 60Co irradiated allogeneic PBMC as feeder cells. Flow cytometry analysis and molecular biology technique were then used to identify γδT cell clones, and MTT assay was used to verify their proliferation after incubated with epitope peptides recognized by γδT cells. Results A γδT cell clone had been established. The subtype of this clone was V γ9 V δ2 and the expression of CD4 and CD8 molecule were negative. Further studies indicated that epitope peptide EP6 could not only specifically bind γδT cell clone but also trigger the proliferation of γδT cell clone. Conclusion A healthy donor's γδT cell clone was successfully established, which laid a solid foundation for further study on γδT cells.

Key words: γδT cells, clone, limit dilution

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