摘要
目的观察NKC2D配体在人脑胶质瘤细胞(HGCs)的表达及其对自然杀伤(NK)细胞杀伤HGC活性的影响。方法流式细胞仪检测HGCs表面的NKG2D配体的表达;采用MTT法检测NK细胞在不同效靶比时对HGCs的杀伤活性;当效靶比为50∶1时,检测不同单抗对杀伤活性的阻断作用。结果HGCs表达NKG2D的配体MICA、ULBP2、ULBP3,不表达NKG2D的配体MICB、ULBP1。MICA、ULBP2和ULBP3分子在HGC表面的表达率分别为(63.52±6.67)%、(62.01±5.65)%和(51.36±2.38)%。效靶比为10∶1、20∶1、50∶1、100∶1时,杀伤活性分别为(23.58±1.69)%、(37.17±3.45)%、(49.69±4.31)%和(54.07±6.12)%;在各效靶比时(除效靶比50∶1组与效靶比100∶1组间外),NK细胞对HGCs的杀伤活性在各组间差异均有统计学意义(P=0.000)。用anti-MICA、anti-ULBP2及anti-ULBP3单抗可明显阻断NK细胞对HGC的杀伤活性,与阻断前相比均有统计学意义(P=0.000)。结论HGCs表达NKG2D的配体MICA、ULBP2、ULBP3,NK细胞在体外可杀伤HGCs。HGCs配体的抗体可明显阻断NK细胞对HGCs的杀伤活性。
Objective To investigate the expression of NKG2D ligands in human glioma cells(HGCs)and their effects on cytotoxicity of natural killer(NK)cells on HGCs in vitro.Methods Expression of NKG2D ligands on the surface of HGCs was analyzed by flow cytometry.MTT method was used to detect cytotoxicity of NK cells against HGCs at different effector-target ratios.the blocking effect of different monoclonal antibodies on the killing activity were detected at the effector-target ratio of 50∶1.Results The erpression of NKG2D ligand MICA,ULBP2 and ULBP3 were detected on HGCs,and MICB and ULBP1 were not detectable.The expression percentage of MICA,ULBP2 and ULBP3 on HGCs were 63.52±6.67%,62.01±5.65%and 51.36±2.38%,respectively.When the effector-target ratio was 5∶1,20∶1,50∶1 and 100∶1,the killing activity was 23.58±1.69%,37.17±3.45%,49.69±4.31%and 54.07±6.12%,respectively.There were statistically significant differences between each group except the groups between the effector-trget ratios of 50∶1 and that of 100∶1(P<0.05).The killing activity of NK cells on HGCs can be significantly blocked by anti-MICA,anti-ULBP2,and anti-ULBP3 mAbs.It's statistical significant compared to pre-block.Conclusion HGCs express NKG2D ligand MICA,ULBP2 and ULBP3.NK cells can kill HGCs in vitro.The antibody of ligand of HGCs can significant block the cytotoxicity of natural killer(NK)cells on HGCs.
作者
张鹏
韩伟杰
孙红卫
黄德海
ZHANG Peng;HAN Weijie;SUN Hongwei;HUANG Dehai(The First Affiliated Hosptial of Zhengzhou University,Zhengzhou 450052,China;Hean Medical College,Zhengzhou 451191,China)
出处
《中国实用神经疾病杂志》
2020年第7期553-556,共4页
Chinese Journal of Practical Nervous Diseases
基金
河南省教育厅基金项目(编号:18A320059)。
