CTLA-4Ig体外处理血液透析患者单个核细胞对Th1/Th2、Treg/Th17免疫平衡的影响

The Alterations of T Cells Percentage and the Changes of Th17 / Treg Induced by CTLA- 4Ig in the Hemodialysis Patients

目的观察CTLA-4Ig体外处理血液透析患者单个核细胞对Th1/Th2、Treg/Th17免疫平衡的影响。方法银川市第一人民医院肾内科收治的20例终末期肾病患者作为观察组,同期20名健康体检者为对照组,分离获得的单个核细胞以RPMI 1640培养基调配细胞浓度为2×106个·m L-1,观察组和对照组均加入CTLA-4Ig 0.4μL(终浓度为1μg·m L-1),空白组加入等体积的细胞培养液。各组细胞经佛波酯、离子霉素和莫能新共同刺激5h后,收集细胞,用ELISA法检测两组细胞上清IFN-γ、IL-4、IL-17的水平,并用RT-PCR检测单个核细胞的Foxp3和视黄酸相关的孤核受体γt(RORγt)的mRNA表达。结果和对照组相比,观察组加入CTLA-4Ig前细胞上清IFN-γ、IL-4和IL-17水平明显升高(P<0.05),RORγτmRNA表达量明显增高,而Foxp3 mRNA表达量明显降低(P<0.05),CTLA-4Ig处理后两组上述指标差异无统计学意义(P>0.05)。观察组CTLA-4Ig处理后IFN-γ、IL-4和IL-17水平均降低(P<0.05或<0.01);Foxp3mRNA表达量明显升高(P<0.01),而RORγτmRNA表达量明显降低(P<0.01)。相关分析显示:IFN-γ、IL-4、IL-17与RORγτ/β-actin相关系数分别为0.985、0.846、0.912(P均<0.01),IFN-γ、IL-4、IL-17与Foxp3/β-actin相关系数分别为-0.983、-0.879、-0.922(P均<0.01)。结论终末期肾病患者微炎症状态下存在Th1/Th2、Treg/Th17失衡,且两者间存在明显的相关性;CTLA-4Ig处理可改善微炎症状态下Th1/Th2、Treg/Th17的失衡。

Objective To explore and analyze the changes of T cell percentage and Treg / Th17 cells induced by CTLA- 4Ig in the hemodialysis patients. Methods 20 healthy subjects and 20 patients with end- stage renal disease were selected as the controls and observed groups,respectively. The obtained mononuclear cells were diluted to 2 × 106 cells / m L by RPMI 1640,and then 0. 4μL CTLA4 Ig was joined into the observed group and the control group while the equal volume of nutrient solution was joined into the blank control group. The levels of IFN- γ,IL- 4,IL- 17 in the two groups patients of harvested culture supernatants were determined by ELISA. Foxp3 and RORγt mRNA level of the patients were detected by fluorescence semi- quantitative PCR. Results Before treatment,IFN- γ,IL- 4 and IL- 17 level of the observed group were significantly higher than those of controls( P < 0. 05),while no significant difference was found between two groups after the intervention. Meanwhile,the RORγt expression quantity of the observed group increased significantly as well as that of Foxp3 decreased significantly before treatment( P < 0. 05),but no significant changes were found after treatment. Compared with the results before treatment,RORγt expression quantity of the observed group decreased( P < 0. 01),while that of the Foxp3 increased after intervention( P < 0. 01). The related analysis showed that the correlation coefficients between IFN- γ,IL- 4,IL- 17 and RORγτ / β- actin were0. 985,0. 846,0. 912,respectively( P < 0. 01),as well as that between IFN- γ,IL- 4,IL- 17 and Foxp3 /β- actin were- 0. 983,- 0. 879,- 0. 922,respectively( P < 0. 01). Conclusion There were imbalance between Th1 and Th2,as well as Treg and Th17 in end- stage renal disease patients with slight inflammation.And the imbalance of Th1 and Th2 was significantly correlated with that of Treg and Th17. In addition,CTLA- 4Ig could improve the imbalance of Th1 / Th2 and Treg / Th17.