■γδT细胞用于肿瘤细胞免疫治疗已成为近期研究的热点。
■为了研究体外扩增的γδT细胞对肝癌细胞的杀伤作用及其机制,和体内验证。
■分离并扩增外周血单核细胞(PBMC)。使用流式细胞术确定T细胞中γδT细胞的比例。选择γδT细胞作为效应细胞,并以HepG2细胞为靶细胞进行细胞毒性实验。NKG2D阻断剂用于阻断效应细胞识别靶细胞,PD98059用于阻断细胞内信号通路。分两批建立裸鼠肿瘤模型,绘制肿瘤生长曲线,并利用小动物成像仪测试肿瘤形成效果,以验证γδT细胞的杀伤作用。
■三个实验组的γδT细胞均表现出大量扩增(P<0.01)。在杀人实验中,实验组唑来膦酸(ZOL)对γδT细胞的杀伤率明显高于HDMAPP组和结核分枝杆菌H37Ra(Mtb-Hag)组(P<0.05)。PD98059的阻断作用强于NKG2D阻断剂(P<0.05)。其中,在HDMAPP组中,当目标比例为40:1时,NKG2D阻断剂表现出明显的阻断作用(P<0.05)。或者,在ZOL组,当效应比为10:1时,用PD98059治疗后效应细胞明显阻断(P<0.05)。体内实验验证了γδT细胞的杀伤作用。根据肿瘤生长曲线,细胞处理后实验组与对照组比较差异有统计学意义(P<0.05)。
■ZOL具有很高的扩增效率和对杀死肿瘤细胞的积极作用。
UNASSIGNED: γδ T cells for tumor cell immunotherapy has recently become a hot topic.
UNASSIGNED: To investigate the stimulation of expanded γδ T cells in vitro to kill liver cancer cells and its mechanism, and in vivo validation.
UNASSIGNED: Peripheral blood mononuclear cells (PBMCs) were isolated and amplified. The proportion of γδ T cells in T cells was determined using flow cytometry. γδ T cells were selected as effector cells, and HepG2 cells as target cells in the cytotoxicity experiment. NKG2D blocker was used to block effector cells from identifying target cells, and PD98059 was used to block intracellular signaling pathways. The nude mice tumor model was established in two batches, the tumor growth curve was drawn, and the tumor formation effect was tested using small animal imager to verify the killing effect of γδ T cells.
UNASSIGNED: The γδ T cells in the three experimental groups exhibited a large amount of amplification (P < 0.01). In the killing experiment, the killing rate of γδ T cells stimulated by zoledronate (ZOL) in the experimental group was significantly higher than that in the HDMAPP group and the Mycobacterium tuberculosis H37Ra strain (Mtb-Hag) group (P < 0.05). The blocking effect of PD98059 is stronger than that of the NKG2D blocker (P < 0.05). Among them, in the HDMAPP group, when the target ratio was 40:1, the NKG2D blocker exhibited a significant blocking effect (P < 0.05). Alternatively, in the ZOL group, when the effect ratio was 10:1, the effector cells were blocked significantly after treatment using PD98059 (P < 0.05). In vivo experiments verified the killing effect of γδ T cells. According to the tumor growth curve, there was a difference between the experimental and control groups after cell treatment (P < 0.05).
UNASSIGNED: ZOL has high amplification efficiency and a positive effect on killing tumor cells.