MPO-ANCA相关性血管炎患者外周血滤泡辅助性T细胞及白介素-21水平的变化及意义
发布时间：2018-08-08 14:20
【摘要】：背景ANCA相关性血管炎(Antineutrophil cytoplasmic autoantibody associated vasculitis,AAV)是一类累及全身小血管的自身免疫性血管炎,主要包括肉芽肿性多血管炎(Granulomatosis with polyangiitis,GPA)、显微镜下多血管炎(Microscopic polyangiitis,MPA)及嗜酸性肉芽肿性多血管炎(Eosinophilic granulomatosis with polyangiitis,EGPA)。ANCA是该类血管炎的主要血清学标志物,其特异性抗原主要有髓过氧化物酶(Myeloperoxidase,MPO)和蛋白酶-3(Proteinase-3,PR-3),与欧美等国家不同,我国MPO-AAV比PR3-AAV更为常见。此外,MPO-AAV发病过程隐匿不易发现,而且病情进展迅速,受累脏器多,死亡率高,因此加强对MPO-AAV发病机制的研究十分必要。MPO-AAV的确切发病机制尚不明确,研究发现MPO-ANCA可能参与了MPO-AAV发病。MPO-ANCA做为一种自身抗体,由浆细胞产生,B细胞分化为浆细胞及浆细胞分泌抗体的过程受到自身免疫系统的调控。近年来一种定位于淋巴滤泡的新型T细胞亚群,滤泡辅助性T细胞(Follicular helper T cells,Tfh)逐渐被认识。该细胞特异性表达CXC趋化因子受体5(CXC-chemokine receptor 5,CXCR5)、可诱导的共刺激分子(Inducible co-stimulator,ICOS)、程序性死亡因子-1(Programmed cell death protein 1,PD-1)等,并分泌白介素-21(Interleukin,IL-21),主要作用是辅助B细胞分化为成熟的浆细胞及产生抗体。研究发现,Tfh细胞与多种自身免疫病有关,而Tfh在MPO-AAV中的作用尚未见研究报道。目的探讨MPO-AAV患者的外周血滤泡辅助性T细胞及白介素-21水平的变化及其与疾病活动度、临床损害间的关系。方法选择40例初发的、未接受治疗的MPO-AAV患者及30例健康人的新鲜抗凝血,先制备单个核细胞悬液(Peripheral blood mononuclear cells,PBMCs),运用流式细胞术(flow cytometry,FCM)检测血管炎患者和健康人外周血Tfh的表达,将CXCR5+CD4+T细胞定义为循环血Tfh细胞(c Tfh细胞),同时检测c Tfh细胞表面分子ICOS及PD-1表达的细胞百分率及平均荧光强度(Mean fluorescence intensity,MFI)。运用酶联免疫吸附测定(enzyme-linked immunosorbent assay,ELISA)方法检测患者和健康人血清IL-21水平及患者MPO-ANCA水平。详细记录患者的临床指标、脏器损害情况及伯明翰血管炎活动度积分(Birmingham Vasculitis Activity Score-version 3,BVAS-V3)。并比较各指标在血管炎组及健康对照组两组之间的表达差异,分析患者组各指标之间的相关性,并探讨其与血管炎疾病活动度及脏器损害间的关系。结果1.一般临床资料:MPO-AAV组共40例,其中男17例,女23例,年龄23~85(64.43±13.36)岁,病程0.5~240(3[1,48])月,血沉7~140(69.31±36.33)mm/h,C反应蛋白(CRP)0.50~176.92(59.33±49.32)mg/L,BVAS值5~37(18.93±5.89)分;健康对照组共30例,其中男13例,女17例,年龄38~79(62.70±11.30)岁。两组之间性别及年龄分布均无明显差异(p0.05),两组数据具有可比性。2.FCM结果:MPO-AAV组c Tfh、ICOS+c Tfh及PD-1+c Tfh细胞的百分率显著高于健康对照组(25.79%±3.72%vs 19.98%±4.72%,p0.001;1.83%±1.06%vs 0.82%±0.60%,p0.001;10.10%±2.85%vs 8.17%±2.66%,p=0.018)。与健康对照组相比,MPO-AAV组c Tfh细胞表面ICOS与PD-1的表达强度MFI均显著升高(59.16±10.12 vs 48.88±7.36,p0.001;532.71±81.14 vs 449.15±65.99,p0.001);3.ELISA结果:MPO-AAV组外周血血清IL-21水平显著高于健康对照组(Z=-4.12,p=0.005);MPO-AAV组外周血血清MPO-ANCA水平(OD值)为0.352[0.248,0.433];4.MPO-AAV组各检测指标之间的相关性分析:MPO-AAV患者c Tfh细胞百分率分别与血清IL-21及血清MPO-ANCA水平均呈正相关(r=0.497,p0.01;r=0.736,p0.01)。c Tfh细胞ICOS的MFI分别与血清IL-21、MPO-ANCA水平呈显著正相关(r=0.586,p0.01;r=0.607,p0.01)。血清IL-21水平与血清MPO-ANCA水平呈显著正相关(r=0.668,p0.01)。而ICOS+c Tfh、PD-1+c Tfh细胞百分率及c Tfh细胞PD-1的MFI与血清IL-21、MPO-ANCA水平均无相关性。5.MPO-AAV组各检测指标与临床指标之间的相关性分析:MPO-AAV患者BVAS评分别与c Tfh细胞百分率、c Tfh细胞ICOS的MFI及血清MPO-ANCA水平呈正相关(r=0.564,p0.01;r=0.513,p0.01;r=0.358,p0.05)。6.MPO-AAV患者中,有肾脏损害组c Tfh细胞百分率及c Tfh细胞ICOS的MFI均高于无肾脏损害组,差异有统计学意义(p0.05)。有肺损害患者与无肺损害患者相比,各主要实验室检测指标间无明显差异。7.MPO-AAV患者中肾BVAS及肺BVAS分别与各指标之间的相关性分析,结果显示,患者肾脏BVAS与c Tfh细胞及其表面分子ICOS表达强度(MFI)呈正相关(r=0.393,p0.05;r=0.389,p0.05),与其他各指标之间均未见相关性。患者肺部BVAS与所有指标之间均未见相关性。结论1.MPO-AAV患者c Tfh及其表面分子ICOS和PD-1的表达水平、IL-21水平均高于对照组,提示MPO-AAV患者中存在c Tfh细胞数量及功能异常;2.MPO-AAV患者c Tfh、血清IL-21及血清MPO-ANCA水平两两之间均呈正相关,提示c Tfh可能通过辅助B细胞产生致病性抗体MPO-ANCA,参与MPO-AAV的发病,这一过程可能是通过IL-21实现的;3.MPO-AAV患者c Tfh数目升高,而且c Tfh细胞百分率及ICOS的MFI均与BVAS评分呈正相关,提示c Tfh细胞数量及功能可能参与MPO-AAV的发病和病情进展。4.MPO-AAV患者中有肾脏损害与c Tfh细胞之间的研究结果,提示MPO-AAV中c Tfh细胞可能与肾脏损害的发生和发展有关。
[Abstract]:Background ANCA related vasculitis (Antineutrophil cytoplasmic autoantibody associated vasculitis, AAV) is a kind of autoimmune vasculitis involving the small vessels of the whole body, including granulomatous vasculitis (Granulomatosis with polyangiitis, GPA), microvasculitis (Microscopic), and eosinophilic meat under microscope. Eosinophilic granulomatosis with polyangiitis (EGPA).ANCA is the main serological marker of this kind of vasculitis. Its specific antigens mainly include myeloperoxidase (Myeloperoxidase, MPO) and protease -3 (Proteinase-3, PR-3). Unlike countries like Europe and America, our MPO-AAV is more common than those in our country. The pathogenesis is not easy to be found, and the disease is progressing rapidly, many organs are involved and the death rate is high. Therefore, it is necessary to strengthen the study of the pathogenesis of MPO-AAV. It is not clear that the exact pathogenesis of.MPO-AAV is not clear. The study found that MPO-ANCA may be involved in the pathogenesis of MPO-AAV as a kind of autoantibody, produced by plasma cells and B cells differentiated into The process of secreting antibodies in plasma cells and plasma cells is regulated by the autoimmune system. In recent years a new type of T cell subgroup located in lymphoid follicles and follicular auxiliary T cells (Follicular helper T cells, Tfh) are gradually recognized. The cells specifically express CXC chemokine receptor 5 (CXC-chemokine receptor 5, CXCR5), and can induce co - induction. The stimulator (Inducible co-stimulator, ICOS), the programmed death factor -1 (Programmed cell death protein 1, PD-1), and the secretion of interleukin -21 (Interleukin, IL-21). The main function is to assist the differentiated cells to differentiate into mature plasma cells and produce antibodies. Objective to investigate the changes in peripheral blood follicle assisted T cells and interleukin -21 levels in the peripheral blood of MPO-AAV patients and their relationship with the degree of disease activity and clinical damage. Methods 40 cases of primary, untreated MPO-AAV and 30 healthy individuals were selected to prepare mononuclear cell suspension first (Peripheral Blood mononuclear cells, PBMCs), using flow cytometry (flow cytometry, FCM) to detect the expression of Tfh in peripheral blood of patients with vasculitis and healthy people. CXCR5+CD4+T cells are defined as circulating blood Tfh cells (C Tfh cells), and the percentage of cell surface molecules and the average fluorescence intensity of the cells are detected. Intensity, MFI). Using enzyme linked immunosorbent assay (enzyme-linked immunosorbent assay, ELISA) to detect serum IL-21 level and patient MPO-ANCA level in patients and healthy people. The clinical indicators, organ damage and Bermingham vasculitis activity (Birmingham Vasculitis Activity Score-version 3) were recorded in detail. 3) and compared the expression difference between the two groups in the vasculitis group and the healthy control group, analyzed the correlation between the indexes of the patients, and discussed the relationship with the activity of vasculitis and the organ damage. Results 1. general clinical data: 40 cases in group MPO-AAV, including 17 male, 23 female, age 23~85 (64.43 + 13.36), and 0. course of disease 0.. 5~240 (3[1,48]) month, erythrocyte sedimentation rate 7~140 (69.31 + 36.33) mm/h, C reactive protein (CRP) 0.50~176.92 (59.33 + 49.32) mg/L, BVAS 5~37 (18.93 + 5.89), 30 cases in healthy control group, including 13 men, 17 women, age 38~79 (62.70 + 11.30) years. There was no significant difference between sex and age distribution between two groups (P0.05), two group data have comparability results The percentage of C Tfh, ICOS+c Tfh and PD-1+c Tfh cells in the MPO-AAV group was significantly higher than that in the healthy control group (25.79% + 3.72%vs 19.98% + 4.72%, p0.001; 1.83% + 1.06%vs 0.82% + 0.60%, p0.001; 10.10% + 8.17% + 2.66%, respectively). 10.12 vs 48.88 + 7.36, p0.001, 532.71 + 81.14 vs 449.15 + 65.99, p0.001); 3.ELISA results: the serum serum IL-21 level of MPO-AAV group was significantly higher than that of the healthy control group (Z=-4.12, p=0.005); the MPO-ANCA level of the peripheral blood serum in the MPO-AAV group was 0.352[0.248,0.433]. The percentage of Tfh cells was positively correlated with the level of serum IL-21 and serum MPO-ANCA (r=0.497, P0.01; r=0.736, P0.01).C Tfh cell ICOS MFI respectively. The percentage of D-1+c Tfh cells and the MFI of PD-1 in C Tfh cells and serum IL-21, MPO-ANCA level were not related to the correlation analysis between the indexes of.5.MPO-AAV group and the clinical indexes. In 358, P0.05).6.MPO-AAV patients, the percentage of C Tfh cells in the kidney damage group and the MFI of ICOS in C Tfh cells were higher than those in the non renal damage group, the difference was statistically significant (P0.05). There was no significant difference between the patients with lung damage and the non lung injury patients, and there was no significant difference between the main laboratory test indexes of the major laboratory tests, and the renal BVAS and lung BVAS were respectively and each finger in the.7.MPO-AAV patients. The correlation analysis between the markers showed that the renal BVAS of the patient was positively correlated with the ICOS expression intensity (MFI) of the C Tfh cells and their surface molecules (r=0.393, P0.05; r=0.389, P0.05), and had no correlation with the other indexes. There was no correlation between the lung BVAS and all the indexes. The level of expression of PD-1 and the level of IL-21 were higher than that of the control group, suggesting that there was a number of C Tfh cells and abnormal function in MPO-AAV patients, and there was a positive correlation between C Tfh, serum IL-21 and serum MPO-ANCA level in 2.MPO-AAV patients, suggesting that C Tfh may be involved in the pathogenesis of pathogenic antibodies by assisting the cells. The number of C Tfh in patients with 3.MPO-AAV increased, and the percentage of C Tfh cells and the MFI of ICOS were positively correlated with the BVAS score, suggesting that the number and function of C Tfh cells may be involved in the pathogenesis and progression of MPO-AAV. The cells may be related to the occurrence and development of renal damage.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R593.2
本文编号：2172061
[Abstract]:Background ANCA related vasculitis (Antineutrophil cytoplasmic autoantibody associated vasculitis, AAV) is a kind of autoimmune vasculitis involving the small vessels of the whole body, including granulomatous vasculitis (Granulomatosis with polyangiitis, GPA), microvasculitis (Microscopic), and eosinophilic meat under microscope. Eosinophilic granulomatosis with polyangiitis (EGPA).ANCA is the main serological marker of this kind of vasculitis. Its specific antigens mainly include myeloperoxidase (Myeloperoxidase, MPO) and protease -3 (Proteinase-3, PR-3). Unlike countries like Europe and America, our MPO-AAV is more common than those in our country. The pathogenesis is not easy to be found, and the disease is progressing rapidly, many organs are involved and the death rate is high. Therefore, it is necessary to strengthen the study of the pathogenesis of MPO-AAV. It is not clear that the exact pathogenesis of.MPO-AAV is not clear. The study found that MPO-ANCA may be involved in the pathogenesis of MPO-AAV as a kind of autoantibody, produced by plasma cells and B cells differentiated into The process of secreting antibodies in plasma cells and plasma cells is regulated by the autoimmune system. In recent years a new type of T cell subgroup located in lymphoid follicles and follicular auxiliary T cells (Follicular helper T cells, Tfh) are gradually recognized. The cells specifically express CXC chemokine receptor 5 (CXC-chemokine receptor 5, CXCR5), and can induce co - induction. The stimulator (Inducible co-stimulator, ICOS), the programmed death factor -1 (Programmed cell death protein 1, PD-1), and the secretion of interleukin -21 (Interleukin, IL-21). The main function is to assist the differentiated cells to differentiate into mature plasma cells and produce antibodies. Objective to investigate the changes in peripheral blood follicle assisted T cells and interleukin -21 levels in the peripheral blood of MPO-AAV patients and their relationship with the degree of disease activity and clinical damage. Methods 40 cases of primary, untreated MPO-AAV and 30 healthy individuals were selected to prepare mononuclear cell suspension first (Peripheral Blood mononuclear cells, PBMCs), using flow cytometry (flow cytometry, FCM) to detect the expression of Tfh in peripheral blood of patients with vasculitis and healthy people. CXCR5+CD4+T cells are defined as circulating blood Tfh cells (C Tfh cells), and the percentage of cell surface molecules and the average fluorescence intensity of the cells are detected. Intensity, MFI). Using enzyme linked immunosorbent assay (enzyme-linked immunosorbent assay, ELISA) to detect serum IL-21 level and patient MPO-ANCA level in patients and healthy people. The clinical indicators, organ damage and Bermingham vasculitis activity (Birmingham Vasculitis Activity Score-version 3) were recorded in detail. 3) and compared the expression difference between the two groups in the vasculitis group and the healthy control group, analyzed the correlation between the indexes of the patients, and discussed the relationship with the activity of vasculitis and the organ damage. Results 1. general clinical data: 40 cases in group MPO-AAV, including 17 male, 23 female, age 23~85 (64.43 + 13.36), and 0. course of disease 0.. 5~240 (3[1,48]) month, erythrocyte sedimentation rate 7~140 (69.31 + 36.33) mm/h, C reactive protein (CRP) 0.50~176.92 (59.33 + 49.32) mg/L, BVAS 5~37 (18.93 + 5.89), 30 cases in healthy control group, including 13 men, 17 women, age 38~79 (62.70 + 11.30) years. There was no significant difference between sex and age distribution between two groups (P0.05), two group data have comparability results The percentage of C Tfh, ICOS+c Tfh and PD-1+c Tfh cells in the MPO-AAV group was significantly higher than that in the healthy control group (25.79% + 3.72%vs 19.98% + 4.72%, p0.001; 1.83% + 1.06%vs 0.82% + 0.60%, p0.001; 10.10% + 8.17% + 2.66%, respectively). 10.12 vs 48.88 + 7.36, p0.001, 532.71 + 81.14 vs 449.15 + 65.99, p0.001); 3.ELISA results: the serum serum IL-21 level of MPO-AAV group was significantly higher than that of the healthy control group (Z=-4.12, p=0.005); the MPO-ANCA level of the peripheral blood serum in the MPO-AAV group was 0.352[0.248,0.433]. The percentage of Tfh cells was positively correlated with the level of serum IL-21 and serum MPO-ANCA (r=0.497, P0.01; r=0.736, P0.01).C Tfh cell ICOS MFI respectively. The percentage of D-1+c Tfh cells and the MFI of PD-1 in C Tfh cells and serum IL-21, MPO-ANCA level were not related to the correlation analysis between the indexes of.5.MPO-AAV group and the clinical indexes. In 358, P0.05).6.MPO-AAV patients, the percentage of C Tfh cells in the kidney damage group and the MFI of ICOS in C Tfh cells were higher than those in the non renal damage group, the difference was statistically significant (P0.05). There was no significant difference between the patients with lung damage and the non lung injury patients, and there was no significant difference between the main laboratory test indexes of the major laboratory tests, and the renal BVAS and lung BVAS were respectively and each finger in the.7.MPO-AAV patients. The correlation analysis between the markers showed that the renal BVAS of the patient was positively correlated with the ICOS expression intensity (MFI) of the C Tfh cells and their surface molecules (r=0.393, P0.05; r=0.389, P0.05), and had no correlation with the other indexes. There was no correlation between the lung BVAS and all the indexes. The level of expression of PD-1 and the level of IL-21 were higher than that of the control group, suggesting that there was a number of C Tfh cells and abnormal function in MPO-AAV patients, and there was a positive correlation between C Tfh, serum IL-21 and serum MPO-ANCA level in 2.MPO-AAV patients, suggesting that C Tfh may be involved in the pathogenesis of pathogenic antibodies by assisting the cells. The number of C Tfh in patients with 3.MPO-AAV increased, and the percentage of C Tfh cells and the MFI of ICOS were positively correlated with the BVAS score, suggesting that the number and function of C Tfh cells may be involved in the pathogenesis and progression of MPO-AAV. The cells may be related to the occurrence and development of renal damage.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R593.2
【参考文献】
相关期刊论文 前4条
1 桂雅迪;帅宗文;胡子盈;张铭明;武琳琳;陈珊宇;;MPO-ANCA相关性血管炎中抗溶酶体相关膜蛋白-2抗体的作用初探[J];安徽医科大学学报;2016年08期
2 武琳琳;帅宗文;;补体活化旁路途经在抗中性粒细胞胞质抗体相关性血管炎发病机制中作用的研究进展[J];中华临床医师杂志(电子版);2015年22期
3 董大群;王忠永;李海英;邱会芬;王静;付文静;张俊花;安荣真;;系统性红斑狼疮患者外周血滤泡辅助性T细胞水平的检测[J];中国皮肤性病学杂志;2014年10期
4 吕远帆;帅宗文;;抗髓过氧化酶物自身抗体对MPO-ANCA相关性血管炎临床表现的影响[J];安徽医学;2014年02期
相关博士学位论文 前1条
1 陶金辉;滤泡辅助性T细胞与系统性红斑狼疮的相关性研究[D];安徽医科大学;2011年
相关硕士学位论文 前2条
1 张蕾;MPO-ANCA相关性血管炎中性粒细胞CD35、CD55、MPO表达以及外周血、尿Ba水平的变化及意义[D];安徽医科大学;2015年
2 吴珊珊;滤泡辅助性T细胞与强直性脊柱炎的关联研究[D];安徽医科大学;2014年
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