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- Title
Anti-Granulocyte Antibodies (C-ANCA, P-ANCA, GS-ANA) Studied by Confocal Scanning Laser Fluorescence Microscopy, ELISA, and Chemiluminescence Techniques.
- Authors
Skogh, T.; Dahlgren, C.; Holmgren, K.; Peen, E.; Stendahl, O.
- Abstract
Fifty-nine patient sera with antibodies against human polymorphonuclear neutrophil granulocyte (PMN) antigens, as determined primarily by indirect immunofluorescence microscopy (11F) screening, were further analysed by enzyme-linked immunosorbent assays (ELISA), The antibodies were primarily characterized by their immunomorphoiogical staining patterns on ethanol-fined 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 (GSANA) 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 α-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 con taming 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.
- Subjects
IMMUNOGLOBULINS; BLOOD plasma; ENZYME-linked immunosorbent assay; LUMINESCENCE; FLUORESCENCE microscopy; PROSPECTING
- Publication
Scandinavian Journal of Immunology, 1991, Vol 34, Issue 2, p137
- ISSN
0300-9475
- Publication type
Article
- DOI
10.1111/j.1365-3083.1991.tb01530.x