Notch信号通路在成人EB病毒感染所致传染性单核细胞增多症中的作用  

The Role of Notch Signaling Pathway in Adult Patients with Epstein-Barr Virus-induced Infectious Mononucleosis

作　　者：李彧[1] 李连香[2] 高瑛[3] LI Yu;LI Lian-Xiang;GAO Ying(Department of Infectious Diseases,Shaanxi Provincial People’s Hospital,Xi’an 710068,Shaanxi Province,China;Department of Gynecology,Shaanxi Provincial People’s Hospital,Xi’an 710068,Shaanxi Province,China;Department of Hematology,Shaanxi Provincial People’s Hospital,Xi’an 710068,Shaanxi Province,China)

机构地区：[1]陕西省人民医院感染性疾病科,陕西西安710068 [2]陕西省人民医院妇科,陕西西安710068 [3]陕西省人民医院血液内科,陕西西安710068

出　　处：《中国实验血液学杂志》2024年第3期920-926,共7页Journal of Experimental Hematology

基　　金：陕西省卫生健康科研基金(2018A005);西安市科技计划项目[20YXYJ0009(11)];陕西省人民医院科技人才计划项目(2021BJ-26);陕西省人民医院2021年科技发展孵化基金项目(2021YJY-19)。

摘　　要：目的:观察成人传染性单核细胞增多症(IM)患者Notch信号通路分子和Th22细胞的变化,检测抑制Notch信号通路对Th22细胞的调控作用。方法:纳入42例IM患者和21例健康对照者,采集外周血,分离血浆和外周血单个核细胞,酶联免疫吸附试验检测血浆白细胞介素(IL)-17和IL-22水平,流式细胞术检测CD3+CD4+IL-17+Th17细胞和CD3+CD4+IL-22+Th22细胞比例,实时定量PCR法检测Th17转录因子维甲酸相关孤独核受体γt(RORγt)、Th22转录因子芳香烃受体(AhR)及Notch信号通路分子(包括Notch受体、Notch配体、Notch下游分子)mRNA相对表达量。纯化CD4+T细胞,使用γ-分泌酶抑制剂(GSI)刺激培养,检测GSI刺激后细胞增殖、Th17和Th22细胞比例、IL-17和IL-22分泌、转录因子mRNA相对表达量变化。结果:IM组外周血单个核细胞中Notch1和Notch2 mRNA的相对表达量分别为13.58±3.18、4.73±1.16,均明显高于对照组的1.09±0.12、1.07±0.15(均P<0.001),而Notch3和Notch4 mRNA相对表达量在IM组和对照组之间的差异无统计学意义(P>0.05)。IM组Notch配体DLL1和Jagged1 mRNA相对表达量、Notch信号下游分子Hes1、Hes5和Hey1 mRNA相对表达量均高于对照组(均P<0.001)。IM患者Th17和Th22细胞比例分别为5.03%±1.15%、4.48%±1.29%,均高于对照组的4.36%±0.82%、3.83%±0.55%(均P<0.05);血浆IL-17和IL-22水平分别为(301.1±53.82)pg/ml、(101.2±16.45)pg/ml,均高于对照组的(237.2±72.18)pg/ml、(84.75±11.83)pg/ml(均P<0.001);RORγt和AhR mRNA相对表达量分别为1.25±0.22、1.21±0.12,均高于对照组的0.99±0.15、1.04±0.11(均P<0.001)。CD4+T细胞增殖水平、Th17细胞比例、IL-17分泌和RORγt mRNA相对表达量在无GSI刺激组和经GSI刺激组之间的差异无统计学意义(P>0.05)。经GSI刺激后Th22细胞比例、IL-22分泌和AhR mRNA相对表达量较无GSI刺激降低(均P<0.05)。结论:Notch信号通路通过AhR调控IM患者CD4+T细胞分泌IL-22,Notch-AhR-Th22细胞通路可能参与IM发病。Objective:To investigate the changes of Notch signaling molecules and Th22 cells in adult patients with infectious mononucleosis(IM),and assess the regulatory function of Notch signaling inhibition to Th22 cells.Methods:Forty-two IM patients and twenty-one healthy controls were enrolled in this study.Their peripheral blood was collected,from which plasma and peripheral blood mononuclear cells(PBMCs)were isolated.Plasma interleukin(IL)-17 and IL-22 were measured by enzyme-linked immunosorbent assay.The percentages of CD3+CD4+IL-17+Th17 cells and CD3+CD4+IL-22+Th22 cells were investigated by flow cytometry.The mRNA relative levels corresponding to Th17 transcription factor retinoic acid related orphan receptorγt(RORγt),Th22 transcription factor aryl hydrocarbon receptor(AhR),and Notch signaling pathway molecules(including Notch receptors,Notch ligands,Notch downstream molecules)were semi-quantified by real-time PCR.CD4+T cells were purified and stimulated withγ-secretase inhibitor(GSI).Cellular proliferation,Th17 and Th22 percentage,IL-17 and IL-22 secretion,transcription factor mRNA were measured in response to GSI stimulation.Results:The relative expression levels of Notch1 and Notch2 mRNA in PBMCs of IM group were 13.58±3.18 and 4.73±1.16,respectively,which were significantly higher than 1.09±0.12 and 1.07±0.15 in PBMCs of control group(both P<0.001).However,there were no significant differences in relative expression levels of Notch3 and Notch4 mRNA between IM group and control group(P>0.05).The relative expression levels of Notch ligands(including DLL1 and Jagged1)mRNA and Notch downstream molecules(including Hes1,Hes5,and Hey1)were increased in IM group compared with control group(all P<0.001).In IM group,the Th17 and Th22 percentage were 5.03%±1.15%and 4.48%±1.29%,respectively,which were both higher than 4.36%±0.82%and 3.83%±0.55%in control group(both P<0.05).In IM group,the IL-17 and IL-22 level were(301.1±53.82)and(101.2±16.45)pg/ml,respectively,which were both higher than(237.2±72.18)and(84.

关 键 词：EPSTEIN-BARR病毒 传染性单核细胞增多症 NOTCH信号通路 TH22细胞 

分 类 号：R512.7[医药卫生—内科学]