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Improved flow cytometric assessment reveals distinct microvesicle (cell-derived microparticle) signatures in joint diseases
- 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.
- Subjects :
- Male
Pathology
Epidemiology
lcsh:Medicine
Arthritis
Osteoarthritis
030204 cardiovascular system & hematology
0302 clinical medicine
Immunophenotyping
Cell-Derived Microparticles
Synovial Fluid
Molecular Cell Biology
lcsh:Science
Child
0303 health sciences
Multidisciplinary
medicine.diagnostic_test
Microvesicle
10051 Rheumatology Clinic and Institute of Physical Medicine
Middle Aged
Flow Cytometry
Phenotype
10076 Center for Integrative Human Physiology
Child, Preschool
Medicine
Female
Joint Diseases
Research Article
medicine.medical_specialty
Adolescent
Nanoparticle tracking analysis
610 Medicine & health
Rheumatoid Arthritis
1100 General Agricultural and Biological Sciences
Flow cytometry
Autoimmune Diseases
03 medical and health sciences
Rheumatology
1300 General Biochemistry, Genetics and Molecular Biology
Diagnostic Medicine
medicine
Synovial fluid
Humans
Biology
030304 developmental biology
Aged
1000 Multidisciplinary
business.industry
lcsh:R
Infant
medicine.disease
Microvesicles
Arthritis, Juvenile
Microscopy, Electron
Biomarker Epidemiology
Immunology
570 Life sciences
biology
Nanoparticles
lcsh:Q
Clinical Immunology
business
Biomarkers
Cytometry
General Pathology
Subjects
Details
- ISSN :
- 19326203
- Volume :
- 7
- Issue :
- 11
- Database :
- OpenAIRE
- Journal :
- PloS one
- Accession number :
- edsair.doi.dedup.....388b8cb341b89680126412e529a7494a