Vallayyachari Kommoju , Christina Mary Mariaselvam , Sree Nethra Bulusu , Benita Nancy Reni Michael , Chengappa Kavadichanda , Molly Mary Thabah , Vir Singh Negi
{"title":"类风湿性关节炎自体滑液影响体外外周和诱导 T 调节细胞的可塑性和功能","authors":"Vallayyachari Kommoju , Christina Mary Mariaselvam , Sree Nethra Bulusu , Benita Nancy Reni Michael , Chengappa Kavadichanda , Molly Mary Thabah , Vir Singh Negi","doi":"10.1016/j.imlet.2024.106859","DOIUrl":null,"url":null,"abstract":"<div><p>The synovial fluid (SF) microenvironment in rheumatoid arthritis (RA) may alter the stability and function of Tregs. In the present study, we assessed cytokine levels and percentage of Tregs, Tregs expressing CXCR3 (Th1-like Treg), CCR6 (Th17-like Treg) in RA peripheral blood (PB) and RA-SF using fluorescence cytometry. Effect of autologous SF on plasticity and function of RA-PB Tregs (pTregs; CD4<sup>+</sup>CD25<sup>hi</sup>CD127<sup>Lo/−</sup>) and induced vimentin-pulsed Tregs (iTregs<sup>VIM</sup>) was assessed <em>in vitro</em>. Cytokines and percentage of Th1-like and Th17-like Tregs were higher in RA-PB than OA-PB; higher in RA-SF than osteoarthritis (OA)-SF. Compared to OA-SF exposed OA-pTregs, RA-SF exposed RA-pTregs showed higher percentage of Th1-like (11% <em>vs</em> 20%) and Th17-like (16% <em>vs</em> 36%) Tregs; higher <em>T-bet</em> (<em>p</em> = 0.0001), <em>RORγ</em> (<em>p</em> = 0.0001) and lower <em>FOXP3</em> (<em>p</em> = 0.0001) gene expression; and diminished percentage suppression of autologous T effector cells (36% <em>vs</em> 74%). RA-SF exposed iTregs<sup>VIM</sup> showed increased percentage of Th1-like and Th17-like Tregs compared to iTregs<sup>VIM</sup> exposed to AB serum (8% <em>vs</em> 0.1%; 21% <em>vs</em> 0.1%). IL-2, Tocilizumab and 5-azacytidine reduced the conversion of iTregs<sup>VIM</sup> (8% <em>vs</em> 2.4%; 21% <em>vs</em> 6.9%), when used in combination. To conclude, microenvironment in the RA synovial fluid is possibly responsible for conversion of pTregs into Th-like Tregs and their functional loss. A blockade of cytokine receptors and methyl transferases could inhibit Tregs conversion, providing clinical relevance for future Tregs targeting therapies.</p></div>","PeriodicalId":3,"journal":{"name":"ACS Applied Electronic Materials","volume":null,"pages":null},"PeriodicalIF":4.3000,"publicationDate":"2024-04-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Rheumatoid arthritis autologous synovial fluid affects the plasticity and function of peripheral and induced T regulatory cells in vitro\",\"authors\":\"Vallayyachari Kommoju , Christina Mary Mariaselvam , Sree Nethra Bulusu , Benita Nancy Reni Michael , Chengappa Kavadichanda , Molly Mary Thabah , Vir Singh Negi\",\"doi\":\"10.1016/j.imlet.2024.106859\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>The synovial fluid (SF) microenvironment in rheumatoid arthritis (RA) may alter the stability and function of Tregs. In the present study, we assessed cytokine levels and percentage of Tregs, Tregs expressing CXCR3 (Th1-like Treg), CCR6 (Th17-like Treg) in RA peripheral blood (PB) and RA-SF using fluorescence cytometry. Effect of autologous SF on plasticity and function of RA-PB Tregs (pTregs; CD4<sup>+</sup>CD25<sup>hi</sup>CD127<sup>Lo/−</sup>) and induced vimentin-pulsed Tregs (iTregs<sup>VIM</sup>) was assessed <em>in vitro</em>. Cytokines and percentage of Th1-like and Th17-like Tregs were higher in RA-PB than OA-PB; higher in RA-SF than osteoarthritis (OA)-SF. Compared to OA-SF exposed OA-pTregs, RA-SF exposed RA-pTregs showed higher percentage of Th1-like (11% <em>vs</em> 20%) and Th17-like (16% <em>vs</em> 36%) Tregs; higher <em>T-bet</em> (<em>p</em> = 0.0001), <em>RORγ</em> (<em>p</em> = 0.0001) and lower <em>FOXP3</em> (<em>p</em> = 0.0001) gene expression; and diminished percentage suppression of autologous T effector cells (36% <em>vs</em> 74%). RA-SF exposed iTregs<sup>VIM</sup> showed increased percentage of Th1-like and Th17-like Tregs compared to iTregs<sup>VIM</sup> exposed to AB serum (8% <em>vs</em> 0.1%; 21% <em>vs</em> 0.1%). IL-2, Tocilizumab and 5-azacytidine reduced the conversion of iTregs<sup>VIM</sup> (8% <em>vs</em> 2.4%; 21% <em>vs</em> 6.9%), when used in combination. To conclude, microenvironment in the RA synovial fluid is possibly responsible for conversion of pTregs into Th-like Tregs and their functional loss. A blockade of cytokine receptors and methyl transferases could inhibit Tregs conversion, providing clinical relevance for future Tregs targeting therapies.</p></div>\",\"PeriodicalId\":3,\"journal\":{\"name\":\"ACS Applied Electronic Materials\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":4.3000,\"publicationDate\":\"2024-04-25\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"ACS Applied Electronic Materials\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S0165247824000336\",\"RegionNum\":3,\"RegionCategory\":\"材料科学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"ENGINEERING, ELECTRICAL & ELECTRONIC\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"ACS Applied Electronic Materials","FirstCategoryId":"3","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0165247824000336","RegionNum":3,"RegionCategory":"材料科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"ENGINEERING, ELECTRICAL & ELECTRONIC","Score":null,"Total":0}
引用次数: 0
摘要
类风湿性关节炎(RA)的滑膜液(SF)微环境可能会改变Tregs的稳定性和功能。在本研究中,我们使用荧光细胞计数法评估了细胞因子水平以及 RA 外周血(PB)和 RA-SF 中 Treg、表达 CXCR3(Th1 类 Treg)和 CCR6(Th17 类 Treg)的 Treg 百分比。在体外评估了自体 SF 对 RA-PB Tregs(pTregs;CD4+CD25hiCD127Lo/-)和诱导波形蛋白脉冲 Tregs(iTregsVIM)的可塑性和功能的影响。细胞因子以及Th1样和Th17样Tregs的百分比在RA-PB中高于OA-PB;在RA-SF中高于骨关节炎(OA)-SF。与暴露于OA-SF的OA-pTregs相比,暴露于RA-SF的RA-pTregs表现出更高的Th1样(11% vs 20%)和Th17样(16% vs 36%)Tregs百分比;更高的T-bet(p = 0.0001)、RORγ(p = 0.0001)和更低的FOXP3(p = 0.0001)基因表达;以及更低的自体T效应细胞抑制百分比(36% vs 74%)。与暴露于 AB 血清的 iTregsVIM 相比,暴露于 RA-SF 的 iTregsVIM 表现出 Th1 样和 Th17 样 Tregs 百分比增加(8% vs 0.1%;21% vs 0.1%)。IL-2、Tocilizumab 和 5-azacytidine 联合使用可降低 iTregsVIM 的转化率(8% vs 2.4%; 21% vs 6.9%)。总之,RA 滑液中的微环境可能是 pTregs 转化为 Th-like Tregs 及其功能丧失的原因。阻断细胞因子受体和甲基转移酶可抑制Tregs转化,为未来Tregs靶向疗法提供临床意义。
Rheumatoid arthritis autologous synovial fluid affects the plasticity and function of peripheral and induced T regulatory cells in vitro
The synovial fluid (SF) microenvironment in rheumatoid arthritis (RA) may alter the stability and function of Tregs. In the present study, we assessed cytokine levels and percentage of Tregs, Tregs expressing CXCR3 (Th1-like Treg), CCR6 (Th17-like Treg) in RA peripheral blood (PB) and RA-SF using fluorescence cytometry. Effect of autologous SF on plasticity and function of RA-PB Tregs (pTregs; CD4+CD25hiCD127Lo/−) and induced vimentin-pulsed Tregs (iTregsVIM) was assessed in vitro. Cytokines and percentage of Th1-like and Th17-like Tregs were higher in RA-PB than OA-PB; higher in RA-SF than osteoarthritis (OA)-SF. Compared to OA-SF exposed OA-pTregs, RA-SF exposed RA-pTregs showed higher percentage of Th1-like (11% vs 20%) and Th17-like (16% vs 36%) Tregs; higher T-bet (p = 0.0001), RORγ (p = 0.0001) and lower FOXP3 (p = 0.0001) gene expression; and diminished percentage suppression of autologous T effector cells (36% vs 74%). RA-SF exposed iTregsVIM showed increased percentage of Th1-like and Th17-like Tregs compared to iTregsVIM exposed to AB serum (8% vs 0.1%; 21% vs 0.1%). IL-2, Tocilizumab and 5-azacytidine reduced the conversion of iTregsVIM (8% vs 2.4%; 21% vs 6.9%), when used in combination. To conclude, microenvironment in the RA synovial fluid is possibly responsible for conversion of pTregs into Th-like Tregs and their functional loss. A blockade of cytokine receptors and methyl transferases could inhibit Tregs conversion, providing clinical relevance for future Tregs targeting therapies.