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Improved flow cytometric assessment reveals distinct microvesicle (cell-derived microparticle) signatures in joint diseases

Authors :
Bence György
Tamás G. Szabó
Lilla Turiák
Matthew Wright
Petra Herczeg
Zsigmond Lédeczi
Ágnes Kittel
Anna Polgár
Kálmán Tóth
Beáta Dérfalvi
Gergő Zelenák
István Böröcz
Bob Carr
György Nagy
Károly Vékey
Steffen Gay
András Falus
Edit I. Buzás
University of Zurich
Source :
PLoS ONE, PLoS ONE, Vol 7, Iss 11, p e49726 (2012)
Publication Year :
2012

Abstract

INTRODUCTION: Microvesicles (MVs), earlier referred to as microparticles, represent a major type of extracellular vesicles currently considered as novel biomarkers in various clinical settings such as autoimmune disorders. However, the analysis of MVs in body fluids has not been fully standardized yet, and there are numerous pitfalls that hinder the correct assessment of these structures. METHODS: In this study, we analyzed synovial fluid (SF) samples of patients with osteoarthritis (OA), rheumatoid arthritis (RA) and juvenile idiopathic arthritis (JIA). To assess factors that may confound MV detection in joint diseases, we used electron microscopy (EM), Nanoparticle Tracking Analysis (NTA) and mass spectrometry (MS). For flow cytometry, a method commonly used for phenotyping and enumeration of MVs, we combined recent advances in the field, and used a novel approach of differential detergent lysis for the exclusion of MV-mimicking non-vesicular signals. RESULTS: EM and NTA showed that substantial amounts of particles other than MVs were present in SF samples. Beyond known MV-associated proteins, MS analysis also revealed abundant plasma- and immune complex-related proteins in MV preparations. Applying improved flow cytometric analysis, we demonstrate for the first time that CD3(+) and CD8(+) T-cell derived SF MVs are highly elevated in patients with RA compared to OA patients (p = 0.027 and p = 0.009, respectively, after Bonferroni corrections). In JIA, we identified reduced numbers of B cell-derived MVs (p = 0.009, after Bonferroni correction). CONCLUSIONS: Our results suggest that improved flow cytometric assessment of MVs facilitates the detection of previously unrecognized disease-associated vesicular signatures.

Details

ISSN :
19326203
Volume :
7
Issue :
11
Database :
OpenAIRE
Journal :
PloS one
Accession number :
edsair.doi.dedup.....388b8cb341b89680126412e529a7494a