Objective: To investigate the expression and clinical significance of T helper cell 9 (Th9) and its cytokine interleukin 9(IL-9) in peripheral blood of patients with chronic lymphocytic leukemia(CLL).
Methods: A total of 43 newly diagnosed patients with chronic lymphocytic leukemia in the First Affiliated Hospital of Xinjiang Medical University from June 2021 to June 2022 were selected as the case group. The patients were divided into Binet A group (13 cases), Binet B group (20 cases) and Binet C group (10 cases) by Binet staging system, and 20 healthy volunteers who underwent physical examinationin in our hospital in the same period served as control group. The proportion of Th9 cells in peripheral blood was detected by flow cytometry, the expression level of Th9 specific transcription factors PU.1 and IRF4 was detected by Western blot, and the expression level of serum cytokine IL-9 was detected by ELISA. The proportion of Th9, the expression of PU.1, IRF4 and IL-9 in each group were compared, and the correlation between the proportion of Th9, IL-9 and clinicopathological indexes of CLL patients was analyzed.
Results: The proportion of Th9, the expression of PU.1, IRF4 and IL-9 in CLL group were significantly higher than those in control group (P<0.05), the proportion of Th9 and the expression of IL-9 in Binet B and C group were higher than those in Binet A group (P<0.05), but there was no significant difference in the proportion of Th9 cells between Binet B group and C group (P>0.05). The expression of IL-9 in Binet C group was significantly higher than that in Binet B group (P<0.05) . The proportion of Th9 cells and IL-9 were highly expression in patients with β2 microglobulin abnormality, IGHV unmutation, P53 abnormality and hepatosplenic lymph node enlargement(P<0.05), but not related to age and sex (P>0.05). The results of Spearman correlation analysis showed that the proportion of Th9 in patients with CLL was negatively correlated with the lymphocytic account and lymphocyte proportion(rs=-0.32，rs=-0.34). The proportion of Th9 and IL-9 were positively correlated with Binet stage, Rai stage and CLL-IPI Scoring (rs=0.79，rs=0.54，rs=0.58； rs=0.72，rs=0.63，rs=0.45), but not with WBC, CD4+ T cells and CD8+T cells (P>0.05). The proportion of Th9 was positively correlated with IL-9 (rs=0.53).
Conclusion: Th9 cells and IL-9 are abnormally highly expressed in CLL, which is related to the poor prognosis of CLL.
题目: Th9细胞及其细胞因子IL-9在慢性淋巴细胞白血病中 的表达及临床意义.
目的: 探讨辅助性T细胞9(Th9)比例及其细胞因子白介素9（IL-9)在慢性淋巴细胞白血病(CLL)患者外周血中的表达水平及临床意义.
方法: 选取2021年6月至2022年6月于新疆医科大学第一附属医院就诊的43例初诊CLL患者作为病例组，采用Binet分期系统将病例组分为Binet A组(13例)、Binet B组(20例)和Binet C组(10例)，同期在本院体检的20例健康志愿者作为对照组。流式细胞术检测外周血Th9细胞比例，Western blot检测Th9特异性转录因子PU.1和IRF4的表达水平，ELISA检测血清细胞因子IL-9的表达水平。比较各组Th9细胞比例、PU.1、IRF4和IL-9的表达差异；结合患者临床资料分析Th9比例、IL-9与CLL临床病理指标的相关性.
结果: CLL组Th9细胞比例、PU.1、IRF4和IL-9的表达均明显高于对照组（P＜0.05）；组内比较，CLL患者Binet B和C组的Th9细胞比例和IL-9的表达均高于Binet A组（P＜0.05），Th9细胞比例在Binet B组与C组间差异无统计学意义（P＞0.05），IL-9在Binet C组中的表达明显高于Binet B组（P＜0.05）；Th9细胞、IL-9在β2微球蛋白异常、IGHV未突变、TP53异常和肝脾淋巴结肿大患者中高表达，IL-9在肝肿大患者中高表达，而与年龄、性别等因素无关（P＞0.05）。Spearman相关性分析结果显示，CLL患者Th9细胞比例与淋巴细胞计数、淋巴细胞比例呈负相关(rs=-0.32，rs=-0.34)，Th9比例、IL-9与Binet分期、Rai分期和CLL-IPI评分呈正相关(rs=0.79，rs=0.54，rs=0.58； rs=0.72，rs=0.63，rs=0.45)，而与WBC、CD4+T细胞、CD8+T细胞等因素无关（P＞0.05），Th9比例与IL-9呈正相关（rs=0.53）.
结论: Th9细胞和IL-9在CLL中异常高表达，并与CLL不良预后相关.
Keywords: T helper cell 9; chronic lymphocytic leukemia; interleukin-9; prognosis.