γδT细胞对血液肿瘤细胞的体外杀伤活性

发布时间：2018-02-23 12:39

本文关键词： γδT细胞血液肿瘤细胞细胞免疫治疗　出处：《中国肿瘤生物治疗杂志》2017年03期 　论文类型：期刊论文

【摘要】：目的:建立γδT细胞培养体系,探讨γδT细胞对不同血液肿瘤细胞的杀伤活性。方法:选取2016年1月至2016年4月在解放军第307医院造血干细胞移植科收治的4例B细胞淋巴瘤患者和该院5例体检健康志愿者,分别分离其外周血单个核细胞(peripheral blood mononuclear cell,PBMC),用唑来磷酸(zoledronate,Zol)和IL-2体外扩增γδT细胞,采用流式细胞术检测γδT细胞对血液肿瘤细胞Jurkat、THP-1、HL-60、K562、Raji、U-937和RPMI-8226的杀伤活性,比较CIK、NK和γδT三种细胞对K562细胞的杀伤作用,检测γδT细胞IFN-γ、TNF-α的分泌水平及CD107a分子的表达水平。结果:成功在体外扩增外周血γδT细胞;γδT细胞对Jurkat、THP-1、HL-60、K562、U-937和RPMI-8226细胞均有明显的杀伤活性(P0.05);γδT细胞对K562细胞的杀伤活性与NK细胞无统计学差异,但明显强于CIK细胞(P0.01);随着与K562细胞共孵育时间的延长,γδT细胞分泌IFN-γ的水平呈现出时间依赖性增加,TNF-α在8 h后逐渐增加;与K562细胞或HL-60细胞共孵育后,γδT细胞CD107a分子的表达水平均显著增加(P0.01)。结论:体外扩增的γδT细胞对血液肿瘤细胞具有较高的杀伤活性,为血液肿瘤的细胞免疫治疗提供实验依据。
[Abstract]:Objective: to establish the culture system of gamma delta T cells, to investigate the cytotoxicity of gamma delta T cells of different blood tumor cells. Methods: from January 2016 to April 2016 in No.307 Hospital of PLA Department of hematopoietic stem cell transplantation in 4 patients with B cell lymphoma and the 5 healthy volunteers, peripheral blood mononuclear cells were isolated (peripheral blood mononuclear cell, PBMC), with zoledronic acid (zoledronate, Zol) amplification of gamma delta T cells and IL-2 in vitro were detected by flow cytometry in T cells on blood tumor cells Jurkat, THP-1, HL-60, K562, Raji, U-937 and the killing activity of RPMI-8226, CIK, killing three NK and gamma delta T cells on K562 cells and detection of IFN- gamma gamma delta T cells, the expression level of TNF- alpha secretion and CD107a molecules. Results: the successful amplification of T cells in the peripheral blood of gamma delta T cells in vitro; for Jurkat, THP-1, HL-60, K562, and U-937 Have the killing activity of RPMI-8226 cells significantly (P0.05); no significant difference between gamma delta T cells on the cytotoxicity of K562 cells and NK cells, but was stronger than that of CIK cells (P0.01); K562 cells and with prolonging of incubation time, gamma delta T cells secreted IFN- levels showed a time dependent increased TNF- alpha increased gradually in the 8 h; and K562 cells or HL-60 cells were incubated, the expression level of gamma delta T cells CD107a molecules were significantly increased (P0.01). Conclusion: the in vitro amplification of gamma delta T cells have higher cytotoxicity to tumor cells in the blood, to provide experimental basis for blood cell immunity the treatment of cancer.

【作者单位】：解放军第307医院造血干细胞移植科全军造血干细胞研究所;
【基金】：国家高技术研究发展计划(863计划)资助项目(2013AA0200103) 北京市科委首都特色课题基金资助项目(Z161100000516184)~~
【分类号】：R733

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