Brimnes J, Halberg P, Jacobsen S, Wiik A, Heegaard N H
Department of Autoimmunology, Statens Serum Institut, Hvidovre University Hospital, Copenhagen, Denmark.
Clin Exp Immunol. 1997 Nov;110(2):250-6. doi: 10.1111/j.1365-2249.1997.tb08324.x.
The objective of this study was to characterize antigens recognized by neutrophil-specific autoantibodies from patients with RA. Sera from 62 RA patients were screened by indirect immunofluorescence (IIF). Positive sera were further tested by ELISAs for antibodies against various granule proteins and by immunoblotting of electrophoretically separated cell, granule or nuclear extracts. Forty-two sera (68%) reacted with ethanol-fixed neutrophils. In the ELISAs 32% of the 28 medium to strongly IIF-positive sera were negative, while 43% were weakly positive for more than one antigen. Immunoblots of whole neutrophils showed IgG reactions at 25-35 kD, in the 55-kD region, at 80 kD, and at 110 kD. Most sera reacted with more than one band. Except for the 55-kD antigen, none of the antigens appeared in lymphocytes. The most notable reactivity in subcellular fractions was with lactoferrin and with bands of 25-35 kD from nuclei. In conclusion, anti-neutrophil autoantibodies from RA patients recognize different antigens in the cytoplasm and in the nucleus. Lactoferrin is one of the common antigens recognized, but also unknown nuclear antigens of 25-35 kD mol. wt are involved.
本研究的目的是鉴定类风湿关节炎(RA)患者中性粒细胞特异性自身抗体所识别的抗原。通过间接免疫荧光法(IIF)对62例RA患者的血清进行筛查。对阳性血清进一步采用酶联免疫吸附测定(ELISA)检测针对各种颗粒蛋白的抗体,并对电泳分离的细胞、颗粒或核提取物进行免疫印迹分析。42份血清(68%)与乙醇固定的中性粒细胞发生反应。在ELISA中,28份中度至强IIF阳性血清中有32%呈阴性,而43%对一种以上抗原呈弱阳性。全中性粒细胞的免疫印迹显示,在25 - 35 kD、55 kD区域、80 kD和110 kD处有IgG反应。大多数血清与多条条带发生反应。除55 kD抗原外,其他抗原均未在淋巴细胞中出现。亚细胞组分中最显著的反应是与乳铁蛋白以及细胞核中25 - 35 kD的条带发生反应。总之,RA患者的抗中性粒细胞自身抗体识别细胞质和细胞核中的不同抗原。乳铁蛋白是所识别的常见抗原之一,但也涉及分子量为25 - 35 kD的未知核抗原。