银屑病关节炎关节中T淋巴细胞的体外信号蛋白图谱分析
Ex Vivo Signaling Protein Mapping in T Lymphocytes in the Psoriatic Arthritis Joints.
作者信息
Fiocco Ugo, Martini Veronica, Accordi Benedetta, Caso Francesco, Costa Luisa, Oliviero Francesca, Scanu Anna, Felicetti Mara, Frallonardo Paola, Facco Monica, Boso Daniele, Molena Beatrice, Zambello Renato, Ramonda Roberta, Cozzi Franco, Scarpa Raffaele, Basso Giuseppe, Semenzato Gianpietro, Dayer Jean-Michel, Doria Andrea, Punzi Leonardo
机构信息
From 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,
出版信息
J Rheumatol Suppl. 2015 Nov;93:48-52. doi: 10.3899/jrheum.150636.
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.
我们评估了总T细胞中的信号蛋白图谱，以分析调节性T细胞（Treg）和CD4 +效应T细胞（Teff）表型的比例，以及持续性银屑病关节炎（PsA）患者滑液（SF）炎症微环境中各自的白细胞介素6受体α（IL-6Rα）表达。我们的方法是使用多重珠免疫测定法测量SF中的IL-6水平。采用反相蛋白质阵列评估PsA患者SF中总T细胞裂解物中的Janus激酶（JAK）1和JAK2、细胞外调节激酶（ERK）1和2、蛋白激酶Cδ（PKCδ）、信号转导子和转录激活子（STAT）1、STAT3和STAT5磷酸化蛋白。通过流式细胞术对PsA患者的SF和匹配的外周血（PB）以及健康对照（HC）的PB中产生IL-17A和IL-17F的CD4 + IL-17A-F + IL-23 + CD4 + Th细胞（Th17）和CD4 + CD25高细胞内叉头盒转录因子+（FOXP3 +）表型的频率以及Treg和Teff细胞的百分比进行定量。我们的结果如下：在PsA SF样本中，发现PsA SF中总T细胞中JAK1、ERK1 / 2、STAT1、STAT3和STAT5磷酸化蛋白协同增加；其中IL-6水平高于HC的PB。扩增的CD4 + IL-17A-F + IL-23 + Th17、CD4 + CD25 - Teff和CD4 + CD25（高）FoxP3 + Treg亚群，显示出相似水平的增强IL-6Rδ表达，局限于PsA关节。在我们的研究中，通过对PsA关节中T淋巴细胞进行体外信号蛋白图谱分析确定的转录网络图谱揭示了IL-1、IL-6和IL-23信号传导与PsA持续性关节炎症中Th17细胞和CD4 + Tregs分化之间的复杂相互作用。
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