基础医学与临床 ›› 2011, Vol. 31 ›› Issue (11): 1210-1216.

• 研究论文 • 上一篇    下一篇

体外扩增γδ T细胞的肿瘤组织趋向性

康宁1,殷珊珊2,汤龙3,崔莲仙2,何维2   

  1. 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
    2. 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
    3. 中国协和医科大学基础医学院,免疫学系
  • 收稿日期:2011-08-29 修回日期:2011-09-06 出版日期:2011-11-05 发布日期:2011-11-02
  • 通讯作者: 何维 E-mail:heweiimu@public.bta.net.cn
  • 基金资助:
    自然科学基金

Migration of in vitro expanded γδ T cells toward tumor tissue

  • Received:2011-08-29 Revised:2011-09-06 Online:2011-11-05 Published:2011-11-02
  • Contact: Wei HE E-mail:heweiimu@public.bta.net.cn

摘要: 摘要 目的 旨在考察体外扩增的γδ T细胞向结直肠癌细胞迁移的能力。方法 采用密度梯度离心法分离外周血单个核细胞(Peripheral blood mononuclear cells,PBMCs),并利用固相化抗T细胞受体(T cell receptor,TCR)γδ抗体进行两周的体外扩增。对扩增的γδ T进行免疫荧光染色和流式细胞仪分析或流式细胞仪分选。采用Bioplex 200流体芯片系统分析细胞因子和趋化因子的表达情况。采用transwell小室进行趋化试验。结果 体外扩增的γδ T细胞主要表达CCR5和CXCR3两种趋化因子受体。四种结直肠癌细胞系和十种结直肠癌肿瘤组织中存在CCR5和CXCR3配体的表达。体外扩增的γδ T细胞在TCR激活的情况下可以大量产生Th1和Th2型细胞因子,进一步募集γδ T细胞。特别是其产生的干扰素γ(Interferon γ,IFN-γ)能够提高结直肠癌细胞CXCR3配体的表达量,从而进一步促进γδ T细胞的趋近。结论 我们的研究结果为γδ T细胞过继免疫治疗结直肠癌提供了重要指征。

关键词: γδ T细胞, 趋化作用, 趋化因子受体 , 结直肠癌

Abstract: Abstract Objective To investigate the migration tendency of the expanded γδ T cells toward colorectal cancer cells. Methods Peripheral blood mononuclear cells (PBMCs) were isolated by density gradient centrifugation and stimulated by immobilized T cell receptor (TCR) gd specific antibody in vitro for two weeks. The expanded cells underwent immunofluorescence staining and flowcytometry analysis or flow sorting. The cytokines and chemokines expressed by gd T cell were assayed using the Bioplex 200 Suspension Array System. The chemotaxis assays were performed in the transwell chambers. Results Those in vitro-expanded γδ T cells primarily expressed chemokine receptors CCR5 and CXCR3. CCR5 and CXCR3 ligands were detected in four different colorectal cancer cell lines and in ten cases of colorectal cancer. Upon TCR engagement, expanded γδ T cells produced large amounts of Th1 and Th2 cytokines, which further increased γδ T cell accumulation. It is noteworthy that the interferon γ (IFN-γ) produced by the γδ T cells markedly increased the expression of CXCR3 ligands in colorectal cancer cells, which further boosted the migration of the expanded γδ T cells. Conculsion Our data may provide important indications for the use of adoptive γδ T cell-based immunotherapy for the treatment of colorectal cancer.

Key words: γδ T cells, Chemotaxis, Chemokine Receptor, Colorectal Cancer

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