Improved Flow Cytometric Assessment Reveals Distinct Microvesicle (Cell-Derived Microparticle) Signatures in Joint Diseases

Bence György, Tamás G. Szabó, Lilla Turiák, Matthew Wright, Petra Herczeg, Zsigmond Lédeczi, A. Kittel, Anna Polgár, Kálmán Tóth, Beáta Dérfalvi, Gergo Zelenák, István Böröcz, Bob Carr, György Nagy, K. Vékey, Steffen Gay, A. Falus, E. Búzás

Research output: Contribution to journalArticle

74 Citations (Scopus)

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.

Original languageEnglish
Article numbere49726
JournalPLoS One
Volume7
Issue number11
DOIs
Publication statusPublished - 2012

Fingerprint

Cell-Derived Microparticles
joint diseases
synovial fluid
Joint Diseases
Synovial Fluid
rheumatoid arthritis
Juvenile Arthritis
osteoarthritis
arthritis
nanoparticles
Osteoarthritis
Nanoparticles
Electron microscopy
Mass spectrometry
Fluids
Mass Spectrometry
Rheumatoid Arthritis
electron microscopy
Electron Microscopy
mass spectrometry

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Improved Flow Cytometric Assessment Reveals Distinct Microvesicle (Cell-Derived Microparticle) Signatures in Joint Diseases. / György, Bence; Szabó, Tamás G.; Turiák, Lilla; Wright, Matthew; Herczeg, Petra; Lédeczi, Zsigmond; Kittel, A.; Polgár, Anna; Tóth, Kálmán; Dérfalvi, Beáta; Zelenák, Gergo; Böröcz, István; Carr, Bob; Nagy, György; Vékey, K.; Gay, Steffen; Falus, A.; Búzás, E.

In: PLoS One, Vol. 7, No. 11, e49726, 2012.

Research output: Contribution to journalArticle

György, B, Szabó, TG, Turiák, L, Wright, M, Herczeg, P, Lédeczi, Z, Kittel, A, Polgár, A, Tóth, K, Dérfalvi, B, Zelenák, G, Böröcz, I, Carr, B, Nagy, G, Vékey, K, Gay, S, Falus, A & Búzás, E 2012, 'Improved Flow Cytometric Assessment Reveals Distinct Microvesicle (Cell-Derived Microparticle) Signatures in Joint Diseases', PLoS One, vol. 7, no. 11, e49726. https://doi.org/10.1371/journal.pone.0049726
György, Bence ; Szabó, Tamás G. ; Turiák, Lilla ; Wright, Matthew ; Herczeg, Petra ; Lédeczi, Zsigmond ; Kittel, A. ; Polgár, Anna ; Tóth, Kálmán ; Dérfalvi, Beáta ; Zelenák, Gergo ; Böröcz, István ; Carr, Bob ; Nagy, György ; Vékey, K. ; Gay, Steffen ; Falus, A. ; Búzás, E. / Improved Flow Cytometric Assessment Reveals Distinct Microvesicle (Cell-Derived Microparticle) Signatures in Joint Diseases. In: PLoS One. 2012 ; Vol. 7, No. 11.
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AU - György, Bence

AU - Szabó, Tamás G.

AU - Turiák, Lilla

AU - Wright, Matthew

AU - Herczeg, Petra

AU - Lédeczi, Zsigmond

AU - Kittel, A.

AU - Polgár, Anna

AU - Tóth, Kálmán

AU - Dérfalvi, Beáta

AU - Zelenák, Gergo

AU - Böröcz, István

AU - Carr, Bob

AU - Nagy, György

AU - Vékey, K.

AU - Gay, Steffen

AU - Falus, A.

AU - Búzás, E.

PY - 2012

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N2 - 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.

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