论文部分内容阅读
目的建立一个合理的CD4T细胞极化的体外细胞培养方法,研究细胞因子刺激的人脐血CD4T细胞极化时产生IL-17的效应/记忆T细胞(Th17细胞)的表达。方法用TGF-β、抗IL-4、抗IFN-γ、IL-6、IL-1β活化幼稚脐血CD4T细胞,促成Th17细胞表型,IL-23具有维护功能。用流式细胞术分析在Th17极化态和Th0非极化态时CD4T细胞上IL-17和IFN-γ的表达,同时用酶联免疫吸附试验(ELISA)测定细胞培养上清液中IL-17和IFN-γ的水平。结果IL-17+IFN-γ-(Th17)细胞比例在Th17极化态时为(28±2.3)%,非极化态时为(3.2±0.4)%;培养上清液中分泌的IL-17浓度在Th17极化态时为(7350±1530)pg/ml,非极化态时为(218±32)pg/ml,两项指标在Th17极化态时都显著高于非极化态时,差异均有高度统计学意义(P<0.001)。结论多种细胞因子刺激的CD4T细胞极化时促进了IL-17产物的表达,体外Th17极化态的建立为研究IL-17相关的免疫性疾病提供了一个平台。
Objective To establish a reasonable CD4 T cell polarization in vitro cell culture method to study the effect of IL-17 production / memory T cells (Th17 cells) on cytokine-stimulated human umbilical cord blood CD4 T cells polarization. Methods The naive cord blood CD4 T cells were activated by TGF-β, anti-IL-4, anti-IFN-γ, IL-6 and IL-1β to promote the phenotype of Th17 cells. IL- The expression of IL-17 and IFN-γ on CD4 T cells in Th17 polarization state and Th0 non-polarization state was analyzed by flow cytometry. Meanwhile, the expression of IL-17 and IL-17 in the cell culture supernatant was detected by enzyme-linked immunosorbent assay (ELISA) 17 and IFN-γ levels. Results The proportion of IL-17 + IFN-γ- (Th17) cells was (28 ± 2.3)% in the Th17 polarization state and (3.2 ± 0.4)% in the non-polarization state. The secretion of IL- 17 concentrations were (7350 ± 1530) pg / ml in Th17 polarization state and (218 ± 32) pg / ml in unpolarization state, both of which were significantly higher in the Th17 polarization state than in the non-polarization state , The difference was highly statistically significant (P <0.001). Conclusions CD4 + T cells stimulated by various cytokines promote the expression of IL-17 product during polarization. The establishment of Th17 polarized state in vitro provides a platform for the study of IL-17-related immune diseases.