慢性淋巴细胞白血病患者Tfh细胞及其亚群水平的变化和临床意义研究

Characteristics and significance of changes in Tfh cells and their subsets in patients with chronic lymphocytic leukemia

摘要:

目的 初步探讨Tfh细胞及其亚群在慢性淋巴细胞白血病（CLL）患者中的变化特点，并分析其临床意义。方法 选取2021年1月—2023年9月新疆医科大学第一附属医院收治的70例初诊CLL患者为CLL组，另选取该院体检健康人员50例为对照组。应用流式细胞术检测Tfh、Tfh1、Tfh2、Tfh17细胞比例，并分析Tfh细胞PD-1和ICOS的表达水平；实时荧光定量聚合酶链反应检测BCL-6、Blimp-1、IL-21基因表达；Western blotting检测BCL-6、Blimp-1蛋白表达；酶联免疫吸附试验检测血清细胞因子IL-21的水平。结果 CLL组外周血Tfh、Tfh1、PD-1⁺Tfh、ICOS⁺Tfh和PD-1⁺ICOS⁺Tfh细胞比例较对照组增加（P <0.05），亚群比值Tfh1/（Tfh2+Tfh17）比值也升高（P <0.05）。与对照组相比，CLL组BCL-6、Blimp-1、IL-21基因和蛋白相对表达量均升高（P <0.05），BCL-6/Blimp-1比值也升高（P <0.05）。Pearson相关性分析结果显示，BCL-6基因和蛋白表达、BCL-6/Blimp-1比值、IL-21水平与Tfh、Tfh1/（Tfh2+Tfh17）均呈正相关（P <0.05）。临床指标分析结果显示，随着IPI评分分组越靠后，Tfh细胞比例、Tfh1/（Tfh2+Tfh17）比值越高，并且与骨髓中B淋巴细胞呈正相关（P <0.05），与免疫球蛋白呈负相关（P <0.05）。结论 CLL患者外周血Tfh细胞参与疾病发病机制，并且Tfh细胞亚群存在偏向于Tfh1细胞的失衡，Tfh细胞的异常分化可能参与CLL的发病和体液免疫紊乱机制。

Abstract:

Objective To investigate the changes and clinical significance of Tfh cells and their subsets in patients with chronic lymphocytic leukemia(CLL).Methods Seventy newly diagnosed CLL patients admitted to the First Affiliated Hospital of Xinjiang Medical University from January 2021 to September 2023 were selected as the CLL group, with 50 healthy individuals as the control group. The proportion of Tfh, Tfh1, Tfh2 and Tfh17 cells was detected by flow cytometry, and the expression levels of PD-1 and ICOS in Tfh cells were detected. The expression levels of BCL-6, Blimp-1 and IL-21 mRNA were detected by PCR. The protein levels of BCL-6 and Blimp-1 were detected by western blot. The serum levels of cytokine IL-21 were detected by ELISA.Results The proportions of Tfh, Tfh1, PD-1⁺ Tfh, ICOS⁺ Tfh, and PD-1⁺ ICOS⁺ Tfh cells in peripheral blood of CLL patients were significantly increased compared with healthy controls, and the subset ratio Tfh1/(Tfh2 + Tfh17) ratio was also significantly increased (P <0.05). Compared with healthy controls, BCL-6, Blimp-1, and IL-21 levels were significantly increased in the CLL patient group, and the BCL-6/Blimp-1 ratio was also significantly increased (P < 0.05). After correlation analysis, BCL-6 levels, BCL-6/Blimp-1 ratio, and IL-21 levels were positively correlated with Tfh and Tfh1/(Tfh2 + Tfh17) (P < 0.05). Clinical indicator analysis revealed that higher IPI scores were associated with an increased proportion of Tfh cells and a higher Tfh1/(Tfh2 + Tfh17) ratio. Additionally, these parameters were positively correlated with bone marrow B lymphocytes (P < 0.05) and negatively correlated with immunoglobulin levels (P < 0.05).Conclusion Peripheral blood Tfh cells in CLL patients are involved in disease pathogenesis, and there is a subset imbalance skewing towards Tfh1 cells. The abnormal differentiation of Tfh cells may play a role in the pathogenesis of CLL and the mechanisms underlying humoral immune dysregulation.