Skogh T, Dahlgren C, Holmgren K, Peen E, Stendahl O
Department of Medical Microbiology, Faculty of Health Sciences, University of Linköping, Sweden.
Scand J Immunol. 1991 Aug;34(2):137-45. doi: 10.1111/j.1365-3083.1991.tb01530.x.
Fifty-nine patient sera with antibodies against human polymorphonuclear neutrophil granulocyte (PMN) antigens, as determined primarily by indirect immunofluorescence microscopy (IIF) screening, were further analysed by enzyme-linked immunosorbent assays (ELISA). The antibodies were primarily characterized by their immunomorphological staining patterns on ethanol-fixed PMN as judged by conventional IIF microscopy, i.e. anti-neutrophil cytoplasmic antibodies (ANCA) giving a pancytoplasmic granular staining pattern (C-ANCA) or a diffuse perinuclear cytoplasmic pattern (P-ANCA), or granulocyte-specific anti-nuclear antibodies (GS-ANA) producing a homogeneous or peripheral nuclear staining pattern. The three distinct patterns were confirmed by confocal scanning laser IIF microscopy. As antigen substrates in the ELISA tests we used an extract from azurophil PMN granules, myeloperoxidase (MPO), and lactoferrin. As expected, most (but not all) of the C-ANCA positive sera turned out positive in the alpha-ELISA assay. Both P-ANCA and GS-ANA positive sera had high frequencies of antibodies against MPO. Occasional P-ANCA positive sera contained antilactoferrin antibodies. Although P-ANCA and GS-ANA in general probably represent the same type of auto-antibodies, we regard it appropriate to make a distinction between the two patterns, until the existence of 'true' granulocyte-specific ANAs has been ruled out. All sera were analysed for their ability to activate PMN in vitro as judged by the generation of a chemiluminescence (CL) response. Sera containing C-ANCA, as well as sera containing P-ANCA or GS-ANA, showed high frequencies of positive CL tests using 'resting' isolated PMN. The reactions were diminished, but not always abolished, by heat-treatment of the sera.
通过间接免疫荧光显微镜检查（IIF）筛查确定的59份含有抗人多形核中性粒细胞（PMN）抗原抗体的患者血清，进一步采用酶联免疫吸附测定（ELISA）进行分析。这些抗体主要通过传统IIF显微镜检查判断其在乙醇固定的PMN上的免疫形态学染色模式来表征，即抗中性粒细胞胞浆抗体（ANCA）呈现全胞浆颗粒状染色模式（C-ANCA）或弥漫性核周胞浆模式（P-ANCA），或粒细胞特异性抗核抗体（GS-ANA）产生均匀或周边核染色模式。共聚焦扫描激光IIF显微镜检查证实了这三种不同模式。在ELISA试验中，我们使用嗜天青PMN颗粒提取物、髓过氧化物酶（MPO）和乳铁蛋白作为抗原底物。正如预期的那样，大多数（但不是全部）C-ANCA阳性血清在α-ELISA试验中呈阳性。P-ANCA和GS-ANA阳性血清中抗MPO抗体的频率都很高。偶尔有P-ANCA阳性血清含有抗乳铁蛋白抗体。尽管一般来说P-ANCA和GS-ANA可能代表同一类型的自身抗体，但在排除“真正的”粒细胞特异性抗核抗体存在之前，我们认为区分这两种模式是合适的。通过化学发光（CL）反应的产生来判断所有血清在体外激活PMN的能力。含有C-ANCA的血清，以及含有P-ANCA或GS-ANA的血清，使用“静息”分离的PMN进行CL试验时，阳性频率较高。血清经热处理后，反应减弱，但并非总是完全消除。
