为探讨IL-17A增强脂肪来源间充质干细胞(adipose-derived mesenchymal stem cell,ADSC)对外周血淋巴细胞的免疫调节作用及机制,收集天津市人民医院4例腹部外科术后留取的患者皮下脂肪组织和外周血,分离、培养、鉴定ADSC并将其分为IL-17A预处理(ADSC-17组)及未处理(ADSC组)2组,分别与外周血淋巴细胞在植物血凝素(phytohemagglutinin,PHA)+IL-2刺激条件下共培养.MTT法检测活化淋巴细胞的增殖抑制率;FACS检测CD4+CD25+T淋巴细胞百分比;ELISA检测培养上清液TGF-β1含量.结果显示,分离培养的ADSC与ADSC-17组均高表达分化群(cluster of differentiation,CD)90、CD105和CD73,均具有分化为成骨及成脂细胞的能力.与ADSC组比较,ADSC-17组显著提高活化淋巴细胞的增殖抑制率(均P＜0.01),呈剂量依赖关系.ADSC-17组CD4+CD25+T淋巴细胞百分比及培养上清液TGF-β1含量相比ADSC组均显著升高(均P＜0.05).由此,IL-17A预处理可增强ADSC抑制淋巴细胞增殖及诱导CD4+CD25+T淋巴细胞分化的能力,是诱导淋巴细胞低反应性的有效策略.
Regulation of lymphocyte function by IL-17A preconditioned adipose-derived mesenchymal stem cells
The aim of this study is to investigate the immuno-modulatory effect of IL-17A preconditioned(ameliorated)adipose-derived mesenchymal stem cells(ADSCs)on peripheral blood lymphocytes and to explore the underlying mechanism.Subcuta-neous adipose tissue was collected from four abdominal surgery patients consulting Tianjin People's Hospital,along with the peripheral blood.ADSCs were isolated,cultured and identified.Next,ADSCs were divided into two groups preconditioned with or without IL-17A(named ADSC-17 group and ADSC group,respectively),which were co-cultured with phytohemagglutinin(PHA)+IL-2-stimulated lymphocytes.The proliferation inhibitory rate of activated lymphocytes was analyzed by MTT assay.The percentage of CD4+CD25+T lymphocytes was detected by FACS.ELISA was performed to detect the TGF-β1 level in cul-ture supernatants.The results revealed that after in vitro culturing,both ADSC and ADSC-17 groups strongly expressed cluster of differentiation(CD)90,CD105,and CD73 as well as displayed osteogenic and adipogenic potentials.The ADSC-17 group showed significantly enhanced proliferation inhibitory rate of activated lymphocytes in a dose-dependent manner compared to that of the ADSC group(all with P＜0.01).The percentage of CD4+CD25+T lymphocytes and the secretion of TGF-β1 in ADSC-17 group were significantly increased(both P＜0.05)compared to those of the ADSC group.Altogether,the preconditioning of ADSCs by IL-17A can augment the capacity of ADSC to inhibit lymphocyte proliferation and to induce CD4+CD25+T lymphocyte differentiation so as to become an effective strategy for inducing lymphocytes hypo-responsiveness.
NETL
NSTL
万方数据
