再生障碍性贫血患者外周血Th17、CD4^+ CD25^+ Treg细胞的免疫应答研究

Study on the immune response of Th17 and CD4^+ CD25^+ Treg cells in peripheral blood of aplastic anemia patients

目的探究再生障碍性贫血患者外周血TH17、CD4^+ CD25^+ Treg细胞的表达特征及血象影响。方法采集血液标本,进行细胞处理、细胞培养、细胞因子的ELISA检测、流式细胞术检测;将实验结果用Mann-Whitney U秩和检验,进行二分类组间比较;用Kruskal-Wallis H(K)秩和检验,进行多分类趋势分析。结果对照组、NSAA组与SAA组相比,外周血CD4^+ CD25^+ Treg细胞比例依次降低;TH17细胞比例依次升高;差异有统计学意义(P<0.05);TNF-α细胞因子水平组间差异无统计学意义(P>0.05)。SAA组、NSAA组外周血血红蛋白计数与TH17细胞比例均呈负相关(r值分别为-0.09、-0.889,P<0.05)。SAA组、NSAA组外周血血红蛋白计数与CD4^+ CD25^+ Treg细胞比例均呈正相关(r值分别为0.889、0.882,P<0.05)。SAA组、NSAA组外周血血红蛋白计数与血清IL-17细胞因子均呈负相关(r值分别为-0.900、-0.975,P<0.05)。结论 Th17/Treg失衡是再生障碍性贫血发生、发展的关键因素;Th17细胞免疫应答增强,CD4^+ CD25^+ Treg细胞免疫应答减弱,加重了再生障碍性贫血患者骨髓衰竭的严重程度。

Objective To study the expression of Th17 and CD4~＋ CD25~＋ Treg cells in peripheral blood of patients with aplastic anemia and the effect on blood indicators. Methods Blood samples were collected for cell processing,cell culture,enzyme-linked immunosorbent assay（ ELISA） detection of cytokines,and flow cytometry detection; the experiment results,using Mann-Whitney U rank test,were compared between groups using two classification; Kruskal-Wallis H（ K） rank test was conducted for the analysis of multi classification trend. Results The proportion of CD4~＋ CD25~＋ Treg cells in peripheral blood decreased in the order of control group,NSAA group and SAA group,while,the proportion of Th17 cells increased in the same order,with the differences among the groups statistically significant（ P〈0. 05）; but the difference of TNF-alpha cytokine levels was among the groups was not statistically significant（ P〈0. 05）. PBMNC was positively correlated with cytokine IL-17 proportion between the SAA group and the NSAA group（ r =-0. 09,r =-0. 889,P〈0. 05）; in group SAA and group NSAA,there was a positive correlation between hemoglobin count in peripheral blood and CD4~＋ CD25~＋ Treg cells proportion（ r = 0. 889,r = 0. 882,P〈0. 05）,while hemoglobin count in peripheral blood was negatively correlated with serum IL-17 cytokines（ r =-0. 900,r =-0. 975,P〈0. 05）. Conclu sion Th17/Treg imbalance is the key factor for the occurrence and development of aplastic anemia; Th17 cell immune response enhanced while CD4＋Treg cell immune responses to CD25＋decreased,which increased the severity degree of bone marrow failure in patients with aplastic anemia.