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Ex Vivo Signaling Protein Mapping in T Lymphocytes in the Psoriatic Arthritis Joints

Ugo Fiocco1, Veronica Martini1, Benedetta Accordi1

  • 1From the Rheumatology Unit, Department of Medicine DIMED, University of Padua; Hematology and Clinical Immunology Branch, University of Padua; Department of Woman and Child Health, University of Padua, Padua; Rheumatology Unit, Department of Clinical Medicine and Surgery, University Federico II, Naples, Italy; and Faculty of Medicine, University Medical Center, University of Geneva, Geneva, Switzerland.U. Fiocco, MD, Rheumatology Unit, Department of Medicine DIMED, University of Padua; V. Martini, Hematology and Clinical Immunology Branch, University of Padua; B. Accordi, Department of Woman and Child Health, University of Padua; F. Caso, MD, Rheumatology Unit, Department of Medicine DIMED, University of Padua and Rheumatology Unit, Department of Clinical Medicine and Surgery, University Federico II; L. Costa, MD, Rheumatology Unit, Department of Medicine DIMED, University of Padua and Rheumatology Unit, Department of Clinical Medicine and Surgery, University Federico II; F. Oliviero, Rheumatology Unit, Department of Medicine DIMED, University of Padua; A. Scanu, Rheumatology Unit, Department of Medicine DIMED, University of Padua; M. Felicetti, MD, Rheumatology Unit, Department of Medicine DIMED, University of Padua; P. Frallonardo, MD, Rheumatology Unit, Department of Medicine DIMED, University of Padua; M. Facco, Hematology and Clinical Immunology Branch, University of Padua; D. Boso, MD, Hematology and Clinical Immunology Branch, University of Padua; B. Molena, Rheumatology Unit, Department of Medicine DIMED, University of Padua; R. Zambello, MD, Hematology and Clinical Immunology Branch, University of Padua; R. Ramonda, MD, Rheumatology Unit, Department of Medicine DIMED, University of Padua; F. Cozzi, MD, Rheumatology Unit, Department of Medicine DIMED, University of Padua; R. Scarpa, MD, Rheumatology Unit, Department of Clinical Medicine and Surgery, University Federico II; G. Basso, MD, Department of Woman and Child Health, University of Padua; G. Semenzato,

The Journal of Rheumatology. Supplement
|November 3, 2015
PubMed

We assessed signaling protein mapping in total T cells, to analyze the proportions of T regulatory (Treg) and TCD4+ effector (Teff) cell phenotypes, and the respective interleukin 6Rα (IL-6Rα) expression in the inflammatory microenvironment of synovial fluid (SF) of patients with sustained psoriatic arthritis (PsA). Our approach was to measure the IL-6 level in SF using a multiplex bead immunoassay. Reverse-phase protein array was used to assess Janus kinase (JAK) 1 and JAK2, extra-cellular regulated kinase (ERK) 1 and 2, protein kinase Cδ (PKCδ), signal transducer and activator and transcription (STAT) 1, STAT3, and STAT5 phosphoproteins in total T cell lysates from SF of patients with PsA. Frequencies of CD4+IL-17A-F+IL-23+ CD4+ Th cells producing IL-17A and IL-17F (Th17) and CD4+CD25high intracellular forkhead box transcription factor+ (FOXP3+) phenotypes, and the percentage of Treg- and Teff- cells were quantified in SF and matched peripheral blood (PB) of patients with PsA and PB of healthy controls (HC) by flow cytometry. Our results were the following: In PsA SF samples, a coordinate increase of JAK1, ERK1/2, STAT1, STAT3, and STAT5 phosphoproteins was found in total T cells in SF of PsA; where IL-6 levels were higher than in PB from HC. Expanded CD4+IL-17A-F+IL-23+ Th17, CD4+ CD25- Teff- and CD4+CD25(high) FoxP3+Treg subsets, showing similar levels of enhanced IL-6Rδ expression, were confined to PsA joints. In our studies, the transcriptional network profile identified by ex vivo signaling protein mapping in T lymphocytes in PsA joints revealed the complex interplay between IL-1, IL-6, and IL-23 signaling and differentiation of Th17 cells and CD4+Tregs in sustained joint inflammation in PsA.

Keywords:
INTERLEUKIN 6Rα SIGNALINGJAK1/STAT3/STAT5 PHOSPHOPROTEINSPSORIATIC ARTHRITIST REGULATORY (TREG) CELLSTH17 CELLS

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