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

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...

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Bibliographic Details
Published in:PloS one 2012-11, Vol.7 (11), p.e49726
Main Authors: György, Bence, Szabó, Tamás G, Turiák, Lilla, Wright, Matthew, Herczeg, Petra, Lédeczi, Zsigmond, Kittel, Agnes, Polgár, Anna, Tóth, Kálmán, Dérfalvi, Beáta, Zelenák, Gergő, Böröcz, István, Carr, Bob, Nagy, György, Vékey, Károly, Gay, Steffen, Falus, András, Buzás, Edit I
Format: Article
Language:English
Subjects:
Adolescent
Age
Aged
Arthritis
Arthritis, Juvenile - blood
Arthritis, Juvenile - diagnosis
Autoimmune diseases
Biocompatibility
Biology
Biomarkers
Biomarkers - metabolism
Biomedical materials
Blood platelets
Body fluids
CD3 antigen
CD8 antigen
Cell-Derived Microparticles
Child
Child, Preschool
Cytometry
Detergents
Diagnosis
Disease
Electron microscopy
Enumeration
Female
Flow
Flow cytometry
Flow Cytometry - methods
Fluid
Fluids
Humans
Immunophenotyping - methods
Infant
Joint diseases
Joint Diseases - blood
Lymphocytes
Lymphocytes B
Lymphocytes T
Lysis
Male
Mass spectrometry
Mass spectroscopy
Medical research
Medicine
Mens health
Microparticles
Microscopy, Electron - methods
Middle Aged
Mimicry
Multiple sclerosis
Nanoparticles
Nanoparticles - chemistry
Osteoarthritis
Osteoarthritis - blood
Osteoarthritis - diagnosis
Patients
Pediatrics
Phenotype
Phenotyping
Proteins
Rheumatoid arthritis
Rheumatoid factor
Rheumatology
Signatures
Synovial fluid
Synovial Fluid - metabolism
T cells
Womens health
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Summary: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. 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. 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). Our results suggest that improved flow cytometric assessment of MVs facilitates the detection of previously unrecognized disease-associated vesicular signatures.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0049726