未成熟树突状细胞对小鼠胶原诱导性关节炎的治疗作用  

Therapeutic effects of immature dendritic cells on murine collagen-induced arthritis

作　　者：杨骁[1] 彭昊[1] 周建林[1] 方洪松[1] 陈森[1] Yang Xiao Peng Hao Zhou Jianlin Fang Hongsong Chen Sen.(Department of Orthopaedics, Renmin Hospital of Wuhan University, Wuhan 430000, Chin)

机构地区：[1]武汉大学人民医院骨科,武汉430000

出　　处：《中华物理医学与康复杂志》2016年第10期721-725,共5页Chinese Journal of Physical Medicine and Rehabilitation

基　　金：国家自然科学基金资助项目（81301592）

摘　　要：目的探讨未成熟树突状细胞（imDC）对小鼠胶原诱导性关节炎（CIA）的治疗作用及其机制。方法分离、培养小鼠骨髓来源的单核细胞，经细胞因子和脂多糖（LPS）分别诱导为未成熟树突状细胞（imDC）和成熟树突状细胞（mDC），并采用流式细胞技术核对细胞表型。选取6-8周龄雌性BALB／c小鼠30只，鸡Ⅱ型胶原对所有小鼠初次免疫，于初次免疫后第6天按随机数字表法将小鼠分为imDC组、mDC组和磷酸盐缓冲液（PBS）组，每组10只，并分别经尾静脉输注imDC、mDC和PBS；初次免疫后第7天对3组小鼠行加强免疫。于加强免疫后第21天观察3组小鼠的关节变形程度，并检测其关节炎指数（AI）、血清抗炎因子IL-10、TGF-β水平以及脾脏CD4＋CD25＋调节性T细胞（Treg）比例。结果经IL-4和GM—CSF诱导后，骨髓单核细胞CD80、CD86、MHC-Ⅱ的表达率分别为32．3％、25．6％、44．0％，经IL-4、GM—CSF和LPS诱导后，CD80、CD86、MHC—Ⅱ的表达率分别为91．5％、90．9％、94．2％，即imDC和mDC诱导分化成功。加强免疫后第21天，imDC组小鼠AI值为（7．32±1．63）分，显著优于同时间点的mDC组[（13．64±2．02）分]和PBS组[（12．78±1．96）分]。加强免疫后第21天，经ELISA检测，imDC组CIA小鼠血清IL-10和TGF-β以及脾脏CD4＋CD25＋ Treg比例均显著高于同时间点的mDC组和PBS组，差异均有统计学意义（P〈0．01）。结论未成熟树突状细胞可有效抑制小鼠CIA的进展，其作用机制可能与未成熟树突状细胞可通过上调抗炎细胞因子的表达，刺激Treg细胞的增殖，从而诱导免疫耐受有关。Objective To explore the therapeutic effects of immature dendritic cells on collagen-induced arthritis （CIA）. Methods Murine marrow-derived monoeytes were isolated and cultivated with eytokines to gen- erate immature dendritic ceils （imDCs） and with lipopolysaccharide （LPS） to generate mature dendritic cells （mDCs）. The differentiation and phenotypes were confirmed with flow cytometry. Murine models of rheumatoid arthritis （RA） were established with collagen Ⅱ transfusion through the caudal vein. On day 6 after first immunization, subdermal injections of imDC, mDC or PBS were administered, and the mice were divided into three groups according to the injection they had received. On day 7 a second immunization was imposed to fortify CIA. Four weeks after the first immunization the severity of CIA was evaluated using an arthritis index （AI） , serum levels of IL-10 and TGF-β using ELISA, as well as regulatory T cell （Treg） populations using flow cytometry. Results The expression rates of CD80, CD86 and MHC-Ⅱ by DCs induced with rrIL-4 and rrGM-CSF were 32.3%, 25.6% and44.0%, and the rates by DCs induced with rrlL-4, rrGM-CSF and LPS were 91.5%, 90.9% and 94.2%, which identified the differentiation and phenotypes of the imDCs and mDCs. The aveage AI of the imDC group was 7.32±1.63, significantly lower than thoseofthe mDC group （13.64±2.02） and the PBS group （12.78±1.96）. The average serum concentrations of IL-10 and TGF-β in the imDC group were significantly higher than in the mDC and PBS groups. The proportion of Treg in the splenocytes of the imDC group was significantly higher than in the mDC and PBS groups. Conclusion CIA was markedly ameliorated by imDC, possibly through up-regulating the ex-pression of anti-inflammatory cytokines like IL-10 and TGF-βand activating the Treg population, which could lead to immune tolerance.

关 键 词：类风湿关节炎 胶原诱导性关节炎 树突状细胞 免疫耐受 调节性T细胞 细胞因子 

分 类 号：R593.22[医药卫生—内科学]