Moodie F D, Leaker B, Cambridge G, Totty N F, Segal A W
Department of Medicine, Institute of Nephrology and Urology, University College London, England, United Kingdom.
Kidney Int. 1993 Mar;43(3):675-81. doi: 10.1038/ki.1993.97.
Anti-neutrophil cytoplasmic antibodies (ANCA) in sera from patients with clinically proven vasculitis have been described as reacting with proteins present in the granules of human neutrophils. We have studied sera from 59 ANCA positive patients to further characterize the antibody response. In addition to the antigens previously identified in the vasculitic syndromes (myeloperoxidase and serine proteinase 3) the majority of these sera contained antibodies that reacted with a cytosolic extract of neutrophils on Western blots. Nearly 40% of these sera had antibodies directed against a cytosolic protein(s) of molecular mass 48 kD. This protein was purified from neutrophil cytosol by ammonium sulphate fractionation, anion exchange and reverse phase chromatography. Amino acid sequence analysis of a proteolytic fragment of this protein identified it as alpha enolase. The anti-enolase antibodies only recognized the alpha isoform and were present in sera giving either a pANCA or cANCA staining pattern by indirect immunofluorescence. Antibodies to alpha enolase were also found in sera from patients with systemic lupus erythematosus, particularly those with renal disease. We conclude that the antibody response in ANCA positive vasculitis is not restricted to neutrophil granule proteins.
临床确诊的血管炎患者血清中的抗中性粒细胞胞浆抗体（ANCA）已被描述为可与人中性粒细胞颗粒中的蛋白质发生反应。我们研究了59例ANCA阳性患者的血清，以进一步明确抗体反应特征。除了先前在血管炎综合征中鉴定出的抗原（髓过氧化物酶和丝氨酸蛋白酶3）外，这些血清中的大多数含有在蛋白质印迹法中与中性粒细胞胞浆提取物发生反应的抗体。这些血清中近40%含有针对分子量为48 kD的胞浆蛋白的抗体。该蛋白通过硫酸铵分级分离、阴离子交换和反相色谱法从中性粒细胞胞浆中纯化得到。对该蛋白的一个蛋白水解片段进行氨基酸序列分析，确定其为α烯醇化酶。抗烯醇化酶抗体仅识别α同工型，且存在于通过间接免疫荧光呈现pANCA或cANCA染色模式的血清中。系统性红斑狼疮患者的血清中也发现了抗α烯醇化酶抗体，尤其是那些患有肾脏疾病的患者。我们得出结论，ANCA阳性血管炎中的抗体反应并不局限于中性粒细胞颗粒蛋白。
