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Multiparameter Phospho-Flow Analysis of Lymphocytes in Early Rheumatoid Arthritis: Implications for Diagnosis and Monitoring Drug Therapy
- Source :
- PLoS ONE, Vol 4, Iss 8, p e6703 (2009), PLoS ONE
- Publication Year :
- 2009
- Publisher :
- Public Library of Science (PLoS), 2009.
-
Abstract
- Background The precise mechanisms involved in the initiation and progression of rheumatoid arthritis (RA) are not known. Early stages of RA often have non-specific symptoms, delaying diagnosis and therapy. Additionally, there are currently no established means to predict clinical responsiveness to therapy. Immune cell activation is a critical component therefore we examined the cellular activation of peripheral blood mononuclear cells (PBMCs) in the early stages of RA, in order to develop a novel diagnostic modality. Methods and Findings PBMCs were isolated from individuals diagnosed with early RA (ERA) (n = 38), longstanding RA (n = 10), osteoarthritis (OA) (n = 19) and from healthy individuals (n = 10). PBMCs were examined for activation of 15 signaling effectors, using phosphorylation status as a measure of activation in immunophenotyped cells, by flow cytometry (phospho-flow). CD3+CD4+, CD3+CD8+ and CD20+ cells isolated from patients with ERA, RA and OA exhibited activation of multiple phospho-epitopes. ERA patient PBMCs showed a bias towards phosphorylation-activation in the CD4+ and CD20+ compartments compared to OA PBMCs, where phospho-activation was primarily observed in CD8+ cells. The ratio of phospho (p)-AKT/p-p38 was significantly elevated in patients with ERA and may have diagnostic potential. The mean fluorescent intensity (MFI) levels for p-AKT and p-H3 in CD4+, CD8+ and CD20+ T cells correlated directly with physician global assessment scores (MDGA) and DAS (disease activity score). Stratification by medications revealed that patients receiving leflunomide, systemic steroids or anti-TNF therapy had significant reductions in phospho-specific activation compared with patients not receiving these therapies. Correlative trends between medication-associated reductions in the levels of phosphorylation of specific signaling effectors and lower disease activity were observed. Conclusions Phospho-flow analysis identified phosphorylation-activation of specific signaling effectors in the PB from patients with ERA. Notably, phosphorylation of these signaling effectors did not distinguish ERA from late RA, suggesting that the activation status of discrete cell populations is already established early in disease. However, when the ratio of MFI values for p-AKT and p-p38 is >1.5, there is a high likelihood of having a diagnosis of RA. Our results suggest that longitudinal sampling of patients undergoing therapy may result in phospho-signatures that are predictive of drug responsiveness.
- Subjects :
- Adult
Male
CD3
Rheumatology/Imaging and Biomarkers
Immunology/Autoimmunity
lcsh:Medicine
Peripheral blood mononuclear cell
Cell Biology/Cell Signaling
Immunophenotyping
Arthritis, Rheumatoid
Immunology/Leukocyte Signaling and Gene Expression
Humans
Medicine
Cytotoxic T cell
Lymphocytes
Phosphorylation
lcsh:Science
Rheumatology/Rheumatoid Arthritis
Rheumatology/Autoimmunity, Autoimmune, and Inflammatory Diseases
Leflunomide
CD20
Multidisciplinary
biology
business.industry
lcsh:R
Flow Cytometry
medicine.disease
Immunology/Leukocyte Activation
Rheumatoid arthritis
Immunology
biology.protein
Female
lcsh:Q
Drug Monitoring
business
CD8
Research Article
medicine.drug
Subjects
Details
- ISSN :
- 19326203
- Volume :
- 4
- Database :
- OpenAIRE
- Journal :
- PLoS ONE
- Accession number :
- edsair.doi.dedup.....55e8a0871212febaf6de665c3c084082
- Full Text :
- https://doi.org/10.1371/journal.pone.0006703