解放军医学杂志2011，Vol.36Issue(4)：372-375,4.
细胞因子对小鼠脾单个核细胞Th17分化和IL-17分泌的影响
Effect of cytokine on differentiation to Th17 and secretion of IL-17 of mouse spleen mononuclear cells
摘要
Abstract
Objective To investigate the effect of exogenous cytokine TGF-β, IL-6 and IL-23 on the differentiation to Th17 and secretion of IL-l7 of mouse spleen mononuclear cells, and discuss their regulatory mechanism in organ transplantation immunity. Methods Mouse spleen mononuclear cells were stimulated by phorbol myristate acetate (PMA) and ionomycin (Ion), and co-cultured with different cytokines for 72 hours. The spleen mononuclear cells were assigned into 4 groups according to the cytokines with which they were co-cultured : blank group, TGF-β group, TGF-β+IL-6 group and TGF-β+ IL-6+IL-23 group. The ratio of Th17 cells was analyzed using flow cytometry. Expressions of RORγt mRNA and IL-l7 mRNA were analyzed using semi-RT-PCR. IL-l7 protein in supernatant was assayed using ELISA. Results Thl7 cells were not found in blank group and TGF-β group, nor were RORγt mRNA in both groups.There was statistical difference on the ratios of Thl7 cells between TGF-β+ IL-6 group and TGF-β+ IL-6+IL23 group (0.30 ％ ± 0.l4％vs 0.51％±0.11％, P＜0.01), and also on the expressions of RORγt mRNA between TGF-β+ IL-6 group and TGF-β+ IL-6+ IL-23 group (0.2l7±0.038 vs 0.458±0.127, P＜0.01). There was no statistical difference on the expression of IL-l7 mRNA between blank group and TGF-β group (0.868±0.222 vs 0.799±0.205, P＞0.05), while a difference was found between TGF-β+IL-6 group and TGF-β+ IL-6+IL-23 group (1.396±0.359 vs 2.415 ±0.604, P＜0.01). There was no statistical difference on the expression of IL-17 protein between blank group and TGF-β group (90.04±22.56pg/ml vs 125.38±27.58pg/ml, P＞0.05) , while difference was found between TGF-β+IL-6 group and TGF-β+IL-6+IL-23 group (973.32±217.33pg/ml vs 1498.46±317.27pg/ml, P＜0.01) . The expressions of IL-17 mRNA and IL-17 protein were higher in TGF-β+IL-6 group and TGF-β+ IL-6+IL-23 group than in blank and TGF-β groups ( P＜0.01) . Conclusions TGF-β+ IL-6 may promote the differentiation and secretion of IL-l7 in vitro. IL-23 may significantly promote the effect of TGF-β+IL-6.关键词
细胞因子类/脾/单个核细胞/Th17/白细胞介素17分类
医药卫生引用本文复制引用
郑慧丽,李海斌,肖漓,赵薇,杜国盛,王振,石炳毅..细胞因子对小鼠脾单个核细胞Th17分化和IL-17分泌的影响[J].解放军医学杂志,2011,36(4):372-375,4.基金项目
全军医学科研"十一五"计划科技攻关课题(06G115) （06G115）
国家科技支撑计划课题(2008BAI60B04) （2008BAI60B04）
