Roitsch S, Goesswein S, Neurath M, Leppkes M (2018)
Publication Type: Journal article
Publication year: 2018
Book Volume: 51
Pages Range: 288-296
Journal Issue: 6
DOI: 10.1080/08916934.2018.1527317
Background:Anti-neutrophil-cytoplasmic antibodies (ANCA) are auto-antibodies directed against components of neutrophil granulocytes and may be found in various inflammatory conditions, like small-vessel vasculitis or ulcerative colitis (UC). Routine ANCA screening is performed on ethanol-fixed neutrophils using indirect immunofluorescence technique. Yet, how neutrophil granule proteins become available to immunologic presentation is a matter of debate. In recent years, various studies have shown that neutrophils are able to extrude their chromatin decorated with granular proteins as neutrophil extracelullar traps (NETs).Aim: We hypothesized that (I) ANCA immunoreactivity may be found on NETs and (II) NETs may serve as a useful tool in a novel approach for ANCA detection.Methods: Sera from patients suffering from either ANCA-associated vasculitis (n=10), UC (n=30) or sera from patients without diagnosed ANCA-associated diseases (n=20), respectively, were subjected to indirect immunofluorescence and a newly developed method to detect ANCA by flow cytometry employing microbead technology.Results: ANCA-related immunofluorescence was readily detectable on ethanol-fixed NETs, establishing NETs as a structure carrying ANCA target antigens. Moreover, we observed that neutrophils form NETs in response to microbeads and stick to the surface of these beads. Using these NET-coated microbeads in flow cytometry, we were capable of reliably detecting p-ANCA, c-ANCA, and a-ANCA in tested patient sera. UC-related complex DNase-1-sensitive ANCA (NET-ANCA) antigens were also detected on NET-coated microbeads.Conclusion: NET-coated microbeads may be commercially developed as a novel tool for automated ANCA screening assays using flow cytometry.
APA:
Roitsch, S., Goesswein, S., Neurath, M., & Leppkes, M. (2018). Detection by flow cytometry of anti-neutrophil cytoplasmic antibodies in a novel approach based on neutrophil extracellular traps. Autoimmunity, 51(6), 288-296. https://doi.org/10.1080/08916934.2018.1527317
MLA:
Roitsch, Stefan, et al. "Detection by flow cytometry of anti-neutrophil cytoplasmic antibodies in a novel approach based on neutrophil extracellular traps." Autoimmunity 51.6 (2018): 288-296.
BibTeX: Download