Locht H, Skogh T, Wiik A
Department of Autoimmunology, Statens Serum Institute, DK-2300 Copenhagen S, Denmark.
Ann Rheum Dis. 2000 Nov;59(11):898-903. doi: 10.1136/ard.59.11.898.
To study the frequency and distribution of antineutrophil cytoplasmic autoantibodies (ANCA) among patients with reactive arthritis (ReA), rheumatoid arthritis (RA), and ulcerative colitis (UC) using different immunological methods.
Fifty serum samples from patients with reactive arthritis (26 with acute disease and 24 with chronic disease-that is disease of more than one year) were analysed for ANCA with indirect immunofluorescence, enzyme linked immunosorbent assay (ELISA) with six different neutrophil granule proteins as antigens, and immunoblotting on whole neutrophil extract and extracts of azurophil and specific granules. Thirty serum samples from patients with RA and UC served as controls in ELISA and indirect immunofluorescence.
Sixteen per cent of patients with ReA were positive in immunofluorescence compared with 30% of patients with RA, and 70% of patients with UC. Thirty two per cent of patients with ReA were positive in ELISA. Antibodies directed against lactoferrin occurred in 20%, antibodies against bactericidal permeability increasing protein (BPI), elastase, cathepsin G, myeloperoxidase, and proteinase 3 were found in 8%, 2%, 2%, 8%, and 6%, respectively. Overall, 50% of RA sera and 53% of UC sera were positive in one or more ELISA assays, the corresponding figures for antibodies against individual antigens were for RA 7%, 3%, 0%, 13%, 47%, 17% and for UC 13%, 20%, 0%, 23%, 10%, and 17%. In immunoblotting, bands corresponding to lactoferrin and BPI were recognised in 44% and 22% of ReA sera.
Antibodies against neutrophil granule antigens are often found in patients with ReA, primarily among those with chronic disease. The different methods detect various subsets of antibodies, with immunoblotting being the most and immunofluorescence the least sensitive.
采用不同免疫方法研究抗中性粒细胞胞浆自身抗体(ANCA)在反应性关节炎(ReA)、类风湿关节炎(RA)和溃疡性结肠炎(UC)患者中的频率及分布情况。
对50份反应性关节炎患者血清样本(26例急性病患者和24例慢性病患者,即病程超过1年)进行ANCA检测,检测方法包括间接免疫荧光法、以6种不同中性粒细胞颗粒蛋白为抗原的酶联免疫吸附测定(ELISA)以及对全中性粒细胞提取物、嗜天青颗粒提取物和特异性颗粒提取物进行免疫印迹分析。30份RA和UC患者血清样本作为ELISA和间接免疫荧光法的对照。
ReA患者中16%免疫荧光检测呈阳性,RA患者为30%,UC患者为70%。ReA患者中32% ELISA检测呈阳性。针对乳铁蛋白的抗体出现率为20%,针对杀菌通透性增强蛋白(BPI)、弹性蛋白酶、组织蛋白酶G、髓过氧化物酶和蛋白酶3的抗体出现率分别为8%、2%、2%、8%和6%。总体而言,50%的RA血清和53%的UC血清在一项或多项ELISA检测中呈阳性,针对单个抗原的抗体在RA血清中的相应比例分别为7%、3%、0%、13%、47%、17%,在UC血清中分别为13%、20%、0%、23%、10%和17%。免疫印迹分析中,44%的ReA血清可识别出与乳铁蛋白对应的条带,22%可识别出与BPI对应的条带。
ReA患者中常可发现抗中性粒细胞颗粒抗原的抗体,主要见于慢性病患者。不同方法检测出的抗体亚群不同,免疫印迹法最敏感,免疫荧光法最不敏感。