人类白细胞抗原特异性抗体通过活化细胞外调节蛋白激酶信号诱导单核细胞定向分化  

Anti-human leukocyte antigen specific antibody induces monocytes to differentiate into macrophages by activating extracellular regulated protein kinases signal

作　　者：解颖欣 侯君 范琦 吴梦涵 童玲 黄玉华[2] 何军[3] 侯建全[2,4] 魏雪栋[2] Xie Yingxin;Hou Jun;Fan Qi;Wu Menghan;Tong Ling;Huang Yuhua;He Jun;Hou Jianquan;Wei Xuedong(School of Medicine,Soochow University,Suzhou 215006,China;Department of Urology,the First Affiliated Hospital of Soochow University,Suzhou 215031,China;HLA Matching Laboratory,Jiangsu Institute of Blood,the First Affiliated Hospital of Soochow University,Suzhou 215031,China;Department of Urology,Dushu Lake Hospital,Soochow University,Suzhou 215125,China)

机构地区：[1]苏州大学医学部,215006 [2]苏州大学附属第一医院泌尿外科,215031 [3]苏州大学附属第一医院江苏省血液研究所HLA配型实验室,215031 [4]苏州大学附属独墅湖医院泌尿外科,215125

出　　处：《中华实验外科杂志》2022年第5期896-899,共4页Chinese Journal of Experimental Surgery

基　　金：姑苏卫生人才项目(GSWS2019033);国家自然科学基金(82070180);江苏省社会发展重点项目临床前沿技术(BE2019656);苏州大学大学生课外学术科研基金重点项目。

摘　　要：目的探讨肾移植术后抗人类白细胞抗原特异性抗体(HLADSA)诱导人单核细胞向巨噬细胞分化的机制。方法构建人内皮细胞(HAEC)与单核细胞(Mon)体外共培养模型。分为未活化(UT)组、完整HLADSA(Ab)活化组、Ides酶切HLADSA活化组(Fab)组,并添加中和抗体/融合蛋白阻断,流式细胞技术分析HLADSA活化后的HAEC对Mon分化及胞内信号转导的影响。采用方差分析检验。结果Ab活化组Mon胞内细胞外调节蛋白激酶(ERK)1/2T202/Y204磷酸化水平高于UT组(2.163±0.311,F=23.070,MD=-1.163,P<0.01),Fab组ERK仅部分磷酸化(1.211±0.072,F=23.070,MD=-0.211,P<0.05),且低于Ab组(MD=0.952,P<0.05)。U0126可完全阻断Ab和Fab活化组Mon胞内ERK磷酸化(1.045±0.087,F=22.240,MD=0.449,P<0.001;0.992±0.032,F=9.474,MD=0.122,P<0.01)。rPSGL-1-Ig及ICAM-1抗体预处理HAEC均可部分阻断Ab诱导的ERK磷酸化(1.375±0.168,F=11.020,MD=0.410,P<0.01;1.509±0.188,F=11.020,MD=0.277,P<0.01)。单用P-选择素糖蛋白配体1(PSGL-1)抗体、CD18抗体预处理Mon后均可完全抑制Fab活化诱导的ERK磷酸化(1.025±0.035,F=21.050,MD=0.242,P<0.01;0.982±0.031,F=21.050,MD=0.285,P<0.01),而单用CD11a抗体仅能部分抑制(1.117±0.027,F=21.050,MD=0.150,P<0.05),CD11b抗体则无法抑制(1.205±0.033,F=21.050,MD=0.062,P>0.05)。CD64抗体预处理Mon可部分抑制Ab诱导的ERK磷酸化(1.438±0.045,F=26.250,MD=0.415,P<0.05),但当联合使用CD64及CD32抗体时可完全抑制(1.239±0.049,F=26.250,MD=0.614,P<0.01)。结论Ab活化后的HAEC主要通过激活Mon胞内ERK信号诱导后者分化。Objective To reveal the mechanism of the differentiation of human monocytes into macrophages induced by anti-human leukocyte antigen specific antibody(HLADSA)after kidney transplantation.Methods The co-culture model of human endothelial cells(HAECs)and monocytes(Mons)was established in vitro.The co-cultured Mons were divided into three groups:unactivated(UT)group,intact HLADSA(Ab)activated group,Ides DSA activated(Fab)group.The neutralizing antibody and/or fusion protein was added to block the EC activation by Ab or Fab.Flow cytometry was used to analyze the effect of the activated HAEC on Mons differentiation and intracellular signal transduction.One-way ANOVA was performed to data assessment.Results Compared with Mons in UT group,extracellular regulated protein kinases(ERK)1/2T202/Y204 was significantly phosphorylated in Ab activated group(2.163±0.311,F=23.070,MD=-1.163,P<0.01).Mons intracellular ERK in Fab group was only partially phosphorylated(1.211±0.072,F=23.070,MD=-0.211,P<0.05),significantly lower than that in Ab activated group(MD=0.952,P<0.05).U0126 completely blocked the intracellular phosphorylation of ERK in Mons in Ab and Fab activated groups(1.045±0.087,F=22.240,MD=0.449,P<0.001;0.992±0.032,F=9.474,MD=0.122,P<0.01).Both pre-treatment with rPSGL-1-Ig or ICAM-1 antibody on HAECs partially blocked Ab activation-induced phosphorylation of ERK(1.375±0.168,F=11.020,MD=0.410,P<0.01;1.509±0.188,F=11.020,MD=0.277,P<0.01).Mons pretreated with P-selectin glyocoprotein ligand-1(PSGL-1)or CD18 antibody could completely inhibit Fab activation-induced phosphorylation of ERK(1.025±0.035,F=21.050,MD=0.242,P<0.01;0.982±0.031,F=21.050,MD=0.285,P<0.01),but CD11a antibody alone could only partially inhibit it(1.117±0.027,F=21.050,MD=0.150,P<0.05),while CD11b antibody could not(1.205±0.033,F=21.050,MD=0.062,P>0.05).Mons pretreated with CD64 antibody partially inhibited Ab-activated phosphorylation of ERK(1.438±0.045,F=26.250,MD=0.415,P<0.05),but it could be completely inhibited when combined with CD32 antibo

关 键 词：肾移植 人类白细胞抗原抗体 抗体介导的排斥反应 单核细胞 

分 类 号：R392[医药卫生—免疫学]