焦磷酸溴代醇诱导的γ9δ2T淋巴细胞对胃癌细胞株杀伤作用的实验研究

Experimental study on killing effect of bromohydrin pyrophosphate induced γ9δ2T lymphocytes on gastric cancer cell lines

目的:探讨焦磷酸溴代醇(Br HPP)诱导的γ9δ2T淋巴细胞对胃癌细胞株SGC-7901的体外杀伤作用,为胃癌的治疗提供新的模式。方法:分离人外周血单核细胞,采用Br HPP联合白细胞介素2(IL-2)、anti-human TCRγ/δ联合IL-2两种方法进行γ9δ2T淋巴细胞的体外诱导培养,在培养的第10天用流式细胞仪分别检测两种方法培养的细胞γ9δ2TCR等的表达,并对培养细胞上清进行IL-2、肿瘤坏死因子α(TNF-α)、干扰素γ(IFN-γ)等细胞因子检测,最后用两种方法培养的细胞对SGC-7901进行体外杀伤实验。结果:Br HPP联合IL-2培养诱导的细胞γ9δ2TCR有较高表达为61. 60%,其中γ9TCR为65. 10%,δ2TCR为64. 10%; anti-human TCRγ/δ联合IL-2培养诱导的细胞γ9δ2TCR表达率较低为10. 70%,其中γ9TCR为98. 10%,δ2TCR为10. 60%;培养细胞上清IL-2、TNF-α、IFN-γ等细胞因子表达与对照组比较有明显差异(P<0. 01),但两实验组之间比较没有差异(P>0. 05)。Br HPP联合IL-2培养诱导的细胞对SGC-7901的杀伤活性为77. 06%,anti-human TCRγ/δ联合IL-2为78. 80%。结论:Br HPP联合IL-2培养诱导的细胞无论是γ9δ2TCR表达,还是IL-2、TNF-α、IFN-γ等细胞因子的分泌水平,以及对SGC-7901的体外杀伤活性都比较高,证明Br HPP联合IL-2的方法诱导的细胞具备了γ9δ2T淋巴细胞的特征和功能,为γ9δ2T淋巴细胞的进一步研究和应用奠定了基础。

Objective:To investigate the killing effect in vitro of γ9δ2 T lymphocytes induced by bromohydrin pyrophosphate(BrHPP)on a gastric cancer cell line SGC-7901 and provide a new model for the treatment of gastric cancer.Methods:Human peripheral blood mononuclear cells were isolated and induced by BrHPP combined with interleukin 2(IL-2)or anti-human TCRγ/δ combined with IL-2 in vitro.The expression ofγ9 andδ2TCR were detected by flow cytometry in 10 days after induction.Cytokines such as IL-2,tumor necrosis factor(TNF-α),and interferon-γ(IFN-γ)in cell culture supernatant were detected.Finally,SGC-7901 cells were cultured with γ9δ2 T lymphocytes induced by two methods to detect the killing effect in vitro.Results:The expression ofγ9δ2 TCR culture induced by BrHPP combined with IL-2 was high 61.60%,of which γ9TCR and δ2TCR were 65.10%and 64.10%.The expression rate ofγ9δ2 TCR culture induced by anti-human TCRγ/δ combined with IL-2 was low 10.70%,of which γ9TCR and δ2TCR were 98.10%and 10.60%.The expression of IL-2,TNF-α and IFN-γ in culture supernatants was significantly different from that in the control group(P<0.01),but there was no difference between the two experimental groups(P >0.05).The killing activities of γ9δ2 T lymphocytes induced by BrHPP combined with IL-2 and by anti-human TCRγ/δ combined with IL-2 on SGC-7901 were 77.06% and 78.80% respectively.Conclusion:The cells induced by BrHPP combined with IL-2 express high levels of γ9δ2 TCR,secrete more IL-2,TNF-α,IFN-γ and other cytokines,and have higher killing activity on SGC-7901.It is proved that the cells induced by BrHPP combined with IL-2 have the characteristics and functions of γ9δ2 T lymphocytes,which lays a foundation for further research and application of the cells.