Fiocco Ugo, Martini Veronica, Accordi Benedetta, Caso Francesco, Costa Luisa, Oliviero Francesca, Scanu Anna, Facco Monica, Boso Daniele, Gatto Mariele, Felicetti Mara, Frallonardo Paola, Ramonda Roberta, Piva Lucia, Zambello Renato, Agostini Carlo, Scarpa Raffaele, Basso Giuseppe, Semenzato Gianpietro, Dayer Jean-Michel, Punzi Leonardo, Doria Andrea
Rheumatology Unit, Department of Medicine DIMED, University of Padova, Via Giustiniani, 2, 35128, Padova, Italy,
Clin Rheumatol. 2015 Sep;34(9):1571-80. doi: 10.1007/s10067-015-3002-2. Epub 2015 Jul 9.
The objective of the study was to quantify the transcriptional profile, as the main T cell lineage-transcription factors on synovial fluid (SF) T cells, in relation to SF cytokines and T cell frequencies (%) of psoriatic arthritis (PsA) patients. Reverse phase protein array was employed to identify interleukin (IL)-23Rp19-, FOXP3- and related orphan receptor gamma T (RORγt)- protein and Janus associated tyrosine kinases 1 (JAK1), signal transducer and activator and transcription 1 (STAT1), STAT3 and STAT5 phosphoproteins in total T cell lysates from SF of PsA patients. IL-1β, IL-2, IL-6, IL-21 and interferon (INF)-γ were measured using a multiplex bead immunoassay in SF from PsA patients and peripheral blood (PB) from healthy controls (HC). Frequencies of CD4(+)CD25(-), CD4(+)CD25(high) FOXP3(+) and CD4(+)CD25(high) CD127(low) Treg, and either mean fluorescence intensity (MFI) of FOXP3(+) on CD4(+) Treg or MFI of classic IL-6 receptor (IL-6R) α expression on CD4(+)CD25(-) helper/effector T cells (Th/eff) and Treg cells, were quantified in SF of PsA patients and in PB from HC by flow cytometry (FC). In PsA SF samples, IL-2, IL-21 and IFN-γ were not detectable, whereas IL-6 and IL-1β levels were higher than in SF of non-inflammatory osteoarthritis patients. Higher levels of IL-23R-, FOXP3- and RORγt proteins and JAK1, STAT1, STAT3 and STAT5 were found in total T cells from SF of PsA patients compared with PB from HC. Direct correlations between JAK1 Y1022/Y1023 and STAT5 Y694, and STAT3 Y705 and IL6, were found in SF of PsA patients. Increased proportion of CD4(+)CD25(high) FOXP3(+) and CD4(+)CD25(high) CD127(low) Treg cells and brighter MFI of IL-6Rα were observed both on CD4(+)CD25(high)- and CD4(+)CD25(-) T cells in PsA SF. The study showed a distinctive JAK1/STAT3/STAT5 transcriptional network on T cells in the joint microenvironment, outlining the interplay of IL-6, IL-23, IL-1β and γC cytokines in the polarization and plasticity of Th17 and Treg cells, which might participate in the perpetuation of joint inflammation in PsA patients.
本研究的目的是量化银屑病关节炎(PsA)患者滑液(SF)T细胞上主要T细胞谱系转录因子的转录谱,并将其与SF细胞因子及T细胞频率(%)相关联。采用反向蛋白质阵列鉴定PsA患者SF中总T细胞裂解物中的白细胞介素(IL)-23Rp19、叉头框蛋白P3(FOXP3)和相关孤儿受体γT(RORγt)蛋白以及Janus相关酪氨酸激酶1(JAK1)、信号转导子和转录激活子1(STAT1)、STAT3和STAT5磷酸化蛋白。使用多重珠免疫测定法检测PsA患者SF和健康对照(HC)外周血(PB)中的IL-1β、IL-2、IL-6、IL-21和干扰素(INF)-γ。通过流式细胞术(FC)对PsA患者SF和HC PB中CD4(+)CD25(-)、CD4(+)CD25(高)FOXP3(+)和CD4(+)CD25(高)CD127(低)调节性T细胞(Treg)亚群的频率,以及CD4(+)Treg上FOXP3(+)的平均荧光强度(MFI)或CD4(+)CD25(-)辅助/效应T细胞(Th/eff)和Treg细胞上经典IL-6受体(IL-6R)α表达的MFI进行定量分析。在PsA SF样本中,未检测到IL-2、IL-21和IFN-γ,而IL-6和IL-1β水平高于非炎性骨关节炎患者的SF。与HC PB相比,PsA患者SF总T细胞中IL-23R、FOXP3和RORγt蛋白以及JAK1、STAT1、STAT3和STAT5水平更高。在PsA患者的SF中发现JAK1的Y1022/Y1023位点与STAT5的Y694位点、STAT3的Y705位点与IL-6之间存在直接相关性。在PsA SF中,CD4(+)CD25(高)FOXP3(+)和CD4(+)CD25(高)CD127(低)Treg细胞比例增加,且CD4(+)CD25(高)和CD4(+)CD25(-)T细胞上IL-6Rα的MFI增强。该研究显示了关节微环境中T细胞上独特的JAK1/STAT3/STAT5转录网络,勾勒出IL-6、IL-23、IL-1β和γC细胞因子在Th17和Treg细胞极化和可塑性中的相互作用,这可能参与了PsA患者关节炎症的持续存在。