神经母细胞瘤患儿外周血γδ T细胞比例及凋亡变化特点

The Characteristics of Peripheral Blood γδ T Cell Proportion and Its Apoptosis in Children with Neuroblastoma

目的分析研究神经母细胞瘤(NB)患儿外周血中γδT细胞比例以及多种细胞因子水平变化情况,并进行γδT细胞的体外培养,分析其凋亡特点,探讨神经母细胞瘤患儿外周血γδT细胞的变化特点及临床意义。方法通过流式细胞术检测NB患儿外周血中γδT细胞(CD3^+、TCRγδ^+)比例,并与健康儿童比较;通过luminex200液相悬浮芯片系统检测细胞因子表达;探索并优化γδT细胞体外培养条件(PAM+IL-2),培养过程中通过检测Annexin V分析细胞凋亡情况。结果 NB患儿外周血中γδT细胞比例明显升高,患者外周血中多种促生存细胞因子减少;同时我们在体外刺激γδT细胞增殖,发现γδT细胞凋亡增加。结论 NB患儿外周血中γδT细胞呈现病理性升高,并且细胞凋亡增加,其可能与外周血中促生存细胞因子减少相关,增加γδT细胞抗凋亡能力,对于其用于NB临床治疗具有重要意义。

Objective The aim of this study is to investigate the proportion of γδ T cells and concentrations of multiple cytokines in peripheral blood of neuroblastoma patients.Furthermore,we will also stimulate γδ T cell proliferation and analyze the apoptosis of γδ T cells.Methods The proportion of peripheral blood γδ T cells(CD3^+、TCRγδ^+)from children with NB was analyzed through flow cytometry.The concentrations of multiple cytokines were detected by luminex200 system.We then stimulated and cultured ,γδ T cells in the condition of PAM and IL-2,and detected the cell apoptosis by Annexin V staining.Results The γδ T cells proportion of NB patients were up-regulated significantly and cytokines with survival promoting effect in serum were down-regulated.Moreover,we found an increase of the apoptosis of NB patient γδ T cells during in vitro culture.Conclusion Our research revealed a pathological augment of γδ T cell proportion and its apoptosis in NB patients,which might due to the decrease of cytokines with survival promoting effect.With that knowledge,we proposed that enhancing the anti-apoptosis ability of γδ T cell is significant to its application to NB clinic treatment.