CD36抗体引起的胎儿新生儿同种免疫性血小板减少症中人脐静脉内皮细胞通透性作用机理的初步研究  

Preliminary study on HUVEC permeability changes in FNAIT induced by CD36 antibody

作　　者：唐颖怡 陈大伟[1] 夏文杰[1] 徐秀章[1] 叶欣[1] 何博[1] TANG Yingyi;CHEN Dawei;XIA Wenjie;XU Xiuzhang;YE Xin;HE Bo(Guangzhou Blood Center,the Key Medical Laboratory of Guangzhou,Guang-zhou 510095,China.)

机构地区：[1]广州血液中心广州市血液安全重点实验室,广东广州510095

出　　处：《中国输血杂志》2023年第9期766-770,共5页Chinese Journal of Blood Transfusion

基　　金：广东省基础与应用基础研究基金项目(2023A1515010924、2022A1515012096);广东省医学科学技术研究基金项目(B2021251);2023年度市校(院)企联合资助专题(2023A03J0549和2023A03J0552);广州市基础研究计划基础与应用基础研究项目(202201010013);广州市科技计划项目(202201011749);广州市医学重点学科(2021-2023年)项目。

摘　　要：目的探索CD36抗体导致胎儿新生儿同种免疫性血小板减少症(fetal/neonatal alloimmune thrombocytopenia,FNAIT)出现胎儿水肿的发病机制,为临床预防和治疗提供参考。方法利用已经建立的CD36单克隆抗体,与人外周血单个核细胞(human peripheral blood mononuclear cells,PBMC)进行孵育,用ELISA法检测细胞培养上清中细胞因子(TNF-α和IL-1β)的浓度。用富细胞因子的细胞上清与人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVEC)共同孵育,通过检测FITC-albumin的荧光强度,研究内皮细胞通透性的变化。结果经流式细胞术检测发现,CD36单克隆抗体可以与人的单核细胞结合。与同型对照抗体相比,CD36单克隆抗体与人的PBMC孵育后,细胞培养上清中检测到细胞因子TNF-α(pg/mL)(407.73±20.40 vs 29.38±4.72,P<0.05)和IL-1β(pg/mL)(247.14±83.59 vs 53.68±26.96,P<0.05)浓度升高。检测HUVEC培养transwell下室中的FITC-albumin荧光强度发现,CD36单克隆抗体与人PBMC孵育后的富细胞因子的细胞培养上清可以显著增加内皮细胞的通透性(CD36抗体vs同型对照抗体MFI值:492±16 vs 320±11,P<0.05)。结论CD36单克隆抗体与PBMC作用可导致HUVEC通透性增加,这可能是FNAIT胎儿水肿的发病机制之一。Objective To explore the pathogenesis of fetal edema caused by CD36 antibody in fetal/neonatal alloimmune thrombocytopenia(FNAIT),and to provide reference for clinical prevention and treatment.Methods The established CD36 monoclonal antibody was incubated with human peripheral blood mononuclear cells(PBMC),and the concentrations of cyto⁃kines(TNF⁃αand IL⁃1β)in the supernatant of cell culture were detected by ELISA.The permeability of endothelial cells were investigated by detecting the fluorescence intensity of FITC⁃albumin by incubating cytokine⁃rich cell supernatant with human umbilical vein endothelial cells(HUVEC).Results Flow cytometry showed that CD36 monoclonal antibody could bind to human monocytes.Compared with isotype IgG control,increased cytokine TNF⁃α(pg/mL)(407.73±20.40 vs 29.38±4.72,P<0.05)and IL⁃1β(pg/mL)(247.14±83.59 vs 53.68±26.96,P<0.05)were detected in the supernatant of cell culture after incubation of CD36 monoclonal antibody with human PBMC.Detection of fluorescence intensity of FITC⁃albumin in transwell cultured HUVEC showed that cytokine⁃rich cell supernatant derived from CD36 monoclonal antibody incubated with human PBMC can increase the permeability of endothelial cells significantly(CD36 antibody vs isotype IgG,MFI value:492±16 vs 320±11,P<0.05).Conclusion The effect of CD36 monoclonal antibody on PBMC can increase HUVEC perme⁃ability,which may be one of the pathogenesis of fetal edema with FNAIT.

关 键 词：CD36抗体 外周血单个核细胞 脐静脉内皮细胞 通透性 胎儿水肿 

分 类 号：R558.2[医药卫生—血液循环系统疾病]