耐受型DC/TGF-β联合诱导的iTreg细胞能有效抑制自身免疫性关节炎  

iTregs induced by tolerogenic DCs plus TGF-β could effectively suppress autoimmune arthritis

作　　者：杨洁[1] 孔宁[2] 杨懿铭[1] 谢如锋[1] 王志成[3] 孙娟[1] 蒋雪玉[1] 

机构地区：[1]上海市血液中心血液工程学科,上海200051 [2]复旦大学附属华山医院风湿职业病科,上海200040 [3]复旦大学附属华山医院检验科,上海200040

出　　处：《安徽医科大学学报》2017年第1期66-72,共7页Acta Universitatis Medicinalis Anhui

基　　金：上海市自然科学基金面上项目(编号:13ZR1438900);国家自然科学基金面上项目(编号:81270650)

摘　　要：目的初步探讨通过成熟耐受型DC(mtDC)与转化生长因子-β(TGF-β)联合诱导的诱导型调节性T细胞(i Treg细胞)对自身免疫性关节炎的抑制作用。方法在体外,对CD4+CD25-T细胞进行TGF-β诱导和mtDC细胞扩增,获得iTreg_(mtDC)细胞。通过流式细胞术、细胞计数、CBA等方法检测iTreg_(mtDC)细胞的表型、对CD4+T效应细胞增殖的抑制作用。在体内实验中,iTreg_(mtDC)细胞以1×10~6个细胞/ml的剂量过继性输注入胶原诱导性关节炎(CIA)小鼠体内,并通过关节病变评分、病理组织病变评分、血清中细胞因子、总anti-CⅡIgG的分泌情况来评价iTreg_(mtDC)细胞对CIA的抑制作用。结果在体外,通过TGF-β和mtDC细胞两轮诱导/扩增后,Treg_(mtDC)细胞得以大量扩增,并能持续表达高水平的Foxp3。在体外,与iTreg细胞相比,Tregmt DC细胞有更强的抑制效应T细胞增殖的能力。在体内,与iTreg细胞相比,Treg_(mtDC)细胞的过继性输注能更有效地减少发病关节处的炎性浸润,进一步改善CIA的症状;对CIA小鼠体内细胞因子的分泌有更强的调节能力,能更显著地减少炎性因子肿瘤坏死因子-α(TNF-α)、白介素-6(IL-6)、白介素-17(IL-17)的分泌,并促进大量TGF-β产生;更明显地降低总anti-CⅡIgG的分泌量;进而更有效地抑制CIA病程的发展。结论与iTreg相比,iTreg_(mtDC)细胞能通过调节体内细胞因子和抗CⅡIgG的分泌更有效地抑制CIA病程发展。Objective To determine the properties of iTregs expanded by mature tolerogenic dendritic cells （mt-DCs） plus TGF-13 and explore their potential toameliorate collagen-induced arthritis （CIA） in a mouse model. Methods In vitro, CD4＋ CD25 - T cells were purified from splenocytes of D1 mice ; after induction by TGF-β and expansion by mtDCs, the phenotype, proliferation and the suppression of iTregmtDCs were assessed by flow cytometry. For in vivo experiments, 1×10^6 iTregmtDC cells were transferred into CIA mice when CIA onset. Clinical and his- topathologie scores, cytokine and anti-CⅡ IgG antibody secretion in serum were analyzed. Results After induction of TGF-β and mtDCs in vitro, the iTregmtDC cells could be expanded effectively and continued to express higher level of Foxp3. Compared with iTregs, iTregmtDC cells showed stronger inhibitory effect. The more remarkable anti-arthritic activity, the improved clinical scores and histological end-points were found in the iTregmtDc-treated group than those in iTreg-treated group. Serological levels of TNF-α, IL-6, IL-17 and anti-C Ⅱ antibodies were also significantly lower and TGF-β production was higher in CIA mice following adoptive transfer of iTregmtDC cells as compared with iTregs. Conclusion These results indicate that iWregmtoC reduced more markedly the severity and progression of CIA than iTregs, which was associated with modulating inflammatory eytokine and anti-CⅡ IgG secretions to lower levels.

关 键 词：iTreg细胞 耐受型树突状细胞 胶原诱导型关节炎 

分 类 号：R593.22[医药卫生—内科学]