细胞间黏附分子-1在CD4^(+)T细胞辅助B细胞产生抗体中的作用和临床意义

Role and clinical significance of intercellular adhesion molecule-1 in CD4^(+)T cells assisting B cells to produce antibodies

目的·探讨细胞间黏附分子-1(intercellular adhesion molecule-1,ICAM-1)在CD4^(+)T细胞辅助B细胞产生抗体中的作用,以及ICAM-1与系统性红斑狼疮(systemic lupus erythematosus,SLE)的相关性。方法·①选取2018年10月—2020年5月在上海交通大学医学院附属仁济医院风湿科就诊的SLE患者50例(SLE组)及仁济医院体检中心健康人60例(健康对照组,即HC组)。采用流式细胞术分析2组样本外周血CD4^(+)T细胞表面ICAM-1表达水平,酶联免疫吸附试验(enzyme-linked immunosorbent assay,ELISA)检测2组样本血清中可溶性ICAM-1(soluble ICAM-1,sICAM-1)含量。②通过密度梯度离心法分离健康人样本外周血单个核细胞,体外经α-CD3/28激活于24、48、72 h采用流式细胞术分析CD4^(+)T细胞表面ICAM-1表达水平,ELISA检测培养上清液中sICAM-1含量。③将健康人CD4^(+)T细胞-B细胞体外共培养,分别设立对照组、刺激组、阻断抗体处理组,12 d后采用ELISA检测3组培养上清液中IgG含量。结果·①SLE组外周血CD4^(+)T细胞表面ICAM-1表达水平以及血清中sICAM-1含量均较HC组升高(均P<0.05)。相关分析显示,CD4^(+)T细胞表面ICAM-1表达水平与红细胞沉降率呈正相关,血清sICAM-1含量与抗双链DNA抗体、IgG水平呈正相关(均P<0.05)。②健康人CD4^(+)T细胞表面ICAM-1表达水平和上清液中sICAM-1含量随着α-CD3/28刺激时间的延长而升高。③共培养后,刺激组上清液中IgG含量较对照组升高,而阻断抗体处理组较刺激组下降(均P<0.05)。结论·ICAM-1分子促进CD4^(+)T细胞-B细胞相互作用后IgG的产生,可作为SLE治疗的潜在靶点。

Objective·To investigate the role of intercellular adhesion molecule-1(ICAM-1)in CD4^(+)T cells assisting B cells to produce antibodies,and the correlation between ICAM-1 and systemic lupus erythematosus(SLE).Methods·①From October 2018 to May 2020,50 SLE patients(SLE group)of Rheumatology Department and 60 healthy people(HC group)of medical examination center from Renji Hospital,Shanghai Jiao Tong University School of Medicine were recruited.Flow cytometry was used to analyze the expression of ICAM-1 on the surface of CD4^(+)T cells in peripheral blood of the two groups.Enzyme-linked immunosorbent assay(ELISA)was used to detect the soluble ICAM-1(sICAM-1)level in serum of the two groups.②Peripheral blood mononuclear cells(PBMCs)of healthy people were isolated by density gradient centrifugation,and activated byα-CD3/28 in vitro at 24,48 and 72 h,respectively.The expression of ICAM-1 on the surface of CD4^(+)T cells was analyzed by flow cytometry,and the level of sICAM-1 in the supernatant was detected by ELISA.③CD4^(+)T cells and B cells of healthy people were co-cultured in vitro,which were divided into control group,stimulation group and antibody blockade group.After 12 days,the IgG level in the co-culture supernatant of the three groups was detected by ELISA.Results·①The expression of ICAM-1 on the surface of CD4^(+)T cells in peripheral blood and the level of sICAM-1 in serum of the SLE group was higher than that of the HC group(both P<0.05).Correlation analysis showed that ICAM-1 expression on the surface of CD4^(+)T cells was positively correlated with erythrocyte sedimentation rate,and serum sICAM-1 level was positively correlated with anti-dsDNA antibody and IgG level(all P<0.05).②The expression of ICAM-1 on the surface of CD4^(+)T cells and the level of sICAM-1 in the supernatant of healthy people were significantly up-regulated gradually along with the stimulationα-CD3/28.③After co-culture,the IgG level in the supernatant of the stimulation group was higher than that in the control group,while IgG lever in antibody blockade group was lower than that in the stimulation group(both P<0.05).Conclusion·ICAM-1 can promote the production of IgG after the interaction of CD4^(+)T cells and B cells,which can be used as a potential target for the treatment of SLE.