DSA对NK细胞介导ADCC效应中血管内皮细胞损伤的影响

Effect of DSA on Vascular Endothelial Cell Injury in NK Cell-Mediated ADCC

目的:探讨抗供者特异性抗体(DSA)在自然杀伤细胞(NK)介导抗体依赖细胞介导的细胞毒效应(ADCC)对人脐静脉内皮细胞(HUVEC)损伤作用。方法:收集10名健康供者为山西医科大学附属肿瘤医院血液科确诊为AML拟行allo-HSCT的患者提供的外周静脉血,并分离提取外周血NK细胞为效应细胞。选取培养至4-6代HUVEC与DSA共培养,将结合DSA的HUVEC作为靶细胞(CDH组),未结合DSA的HUVEC作为阴性对照(UDH组);将效应细胞与靶细胞共培养后通过流式细胞仪检测细胞因子干扰素-γ(IFN-γ)表达,应用MTT法检测HUVEC细胞活性,间接反映DSA介导ADCC效应对内皮细胞损伤作用的强弱。结果:随着效靶比增加,HUVEC细胞活性降低,IFN-γ表达水平增高,在同一效靶比(1∶1、10∶1、20∶1)下,CDH组HUVEC细胞活性明显低于UDH组,IFN-γ表达水平明显高于UDH组;在效靶比为1∶1时,随着DSA浓度提高,CDH组HUVEC细胞OD值渐降低,IFN-γ表达水平增高,差异具有统计学意义(P <0.05)。结论:DSA可通过NK细胞介导的ADCC效应损伤血管内皮细胞。

Objective:To investigate the injury effect of anti-donor specific antibody(DSA)on human umbilical vein enolothelial cells(HUVEC)in NK-mediated antibody-dependent cell cytotoxity(ADCC).Methods:The peripheral blood of 10 healthy donors was colleced for allo-HSCT of AML patients diagnosed in Department of Hemology of the Tumor Hospital affiliated to Shanxi Medical University,then the peripheral blood NK cells were isolated and used as the effector cells;the HUVEC of passages 9-6 were selected and co-cultured with DSA,then the DSA-binding HUVEC were used as the target cells(CDH group),while the DSA-unbinding HUVEC were used as negative control(UDH group).After co-culture of effecor cells with target cells,the expression of IFN-γ was detected by flow cytometry and the HUVEC activity was detected by using MTT method,so as to indirectily reflect the injury effect of DSA-mediated ADCC on endothelial cells.Results:With the increase of effector-target(E:T)ratio,the activity of HUVEC decreased,the expression level of IFN-γ increased.Under the some effector-target ratio(1∶1,10∶1,20∶1),the activity of HUVEC in CDH group was significantly lower than that of UDH group,and the expression of IFN-γ was significantly higher than tmhaetd ioaft etdh eb yU NDKH cgerlolsu.p(P<0.05).Conclusion:DSA can damage vascular endothelial cells through the ADCC effect mediated by NK cells.