慢性淋巴细胞白血病中2型天然淋巴细胞通过白细胞介素-9对调节性T细胞和CD8 T细胞功能的影响
Impact of innate lymphoid cell type 2 in chronic lymphocytic leukemia on the function of treg and CD8 T cells through IL-9.
作者信息
Yang Ruixue, Zeng Xuejiao, Alimu Xierenguli, Qu Jianhua
机构信息
Center of Hematology, The First Affiliated Hospital of Xinjiang Medical University, Hematology Institute of Xinjiang Uygur Autonomous Region, Hematology Clinical Research Center of Xinjiang Uygur Autonomous Region, No.137, Liyu Shan Road, Xinshi District, Urumqi, 830054, China.
出版信息
Cancer Immunol Immunother. 2025 May 30;74(7):228. doi: 10.1007/s00262-025-04082-4.
OBJECTIVE
This study investigated the impact of innate lymphoid cell type 2 (ILC2s) on the function of regulatory T cells (Treg) and CD8 T cells in chronic lymphocytic leukemia (CLL) through IL-9.
METHODS
Peripheral blood samples were collected from CLL patients (n = 52) and healthy controls (n = 30). ILC2 proportions and IL-9 levels were assessed using flow cytometry and ELISA. Immunofluorescence staining was performed to stain GATA3, CRTH2, and IL-9 in cervical lymph nodes from CLL patients (n = 10) and control subjects with reactive lymphadenitis (n = 10). Correlation analysis between ILC2s and IL-9 was conducted using the Spearman test. ILC2s were sorted and cultured from CLL patients, followed by co-culture experiments with PBMCs of healthy controls and MEC-1 cells, with or without anti-IL-9 antibody intervention. Flow cytometry was used to measure the proportions of ILC2s, Treg cells, PD-1/TIGIT/CTLA-4 Treg subsets, and granzyme B/perforin CD8 T cells, along with MEC-1 cell apoptosis.
RESULTS
The proportions of ILC2s and Treg, along with serum IL-9 levels, were significantly elevated in CLL patients (P < 0.05). Peripheral blood ILC2s were positively correlated with IL-9 (r = 0.609, P < 0.001). The average fluorescence intensity of GATA3, CRTH2, and IL-9 in the cervical lymph nodes of CLL patients increased significantly (P < 0.001), and IL-9 showed colocalization with GATA3 and CRTH2. In vitro, IL-9 levels in the supernatant of sorted ILC2s from CLL patients increased. Treatment with anti-IL-9 antibody significantly reduced the PD-1 Treg and TIGIT Treg cells while increasing granzyme B CD8 T cells (P < 0.05). However, there was no significant effect on Treg, CTLA-4 Treg, and perforin CD8 T cells (P > 0.05). Additionally, anti-IL-9 antibody significantly increased early apoptosis (P < 0.05).
CONCLUSION
ILC2s affect CD8 T cells and Treg cells through IL-9, weakening the anti-tumor effects of CD8 T cells and enhancing the immunosuppressive effects of Treg cells, thereby contributing to CLL pathogenesis.
目的
本研究通过白细胞介素-9（IL-9）探讨2型天然淋巴细胞（ILC2s）对慢性淋巴细胞白血病（CLL）中调节性T细胞（Treg）和CD8 T细胞功能的影响。
方法
收集CLL患者（n = 52）和健康对照者（n = 30）的外周血样本。采用流式细胞术和酶联免疫吸附测定法评估ILC2比例和IL-9水平。对10例CLL患者和10例反应性淋巴结炎对照者的颈部淋巴结进行免疫荧光染色，以标记GATA3、CRTH2和IL-9。使用Spearman检验进行ILC2s与IL-9之间的相关性分析。从CLL患者中分离并培养ILC2s，然后与健康对照者的外周血单核细胞（PBMCs）和MEC-1细胞进行共培养实验，有无抗IL-9抗体干预。采用流式细胞术检测ILC2s、Treg细胞、PD-1/TIGIT/CTLA-4 Treg亚群以及颗粒酶B/穿孔素CD8 T细胞的比例，以及MEC-1细胞凋亡情况。
结果
CLL患者中ILC2s和Treg的比例以及血清IL-9水平显著升高（P < 0.05）。外周血ILC2s与IL-9呈正相关（r = 0.609，P < 0.001）。CLL患者颈部淋巴结中GATA3、CRTH2和IL-9的平均荧光强度显著增加（P < 0.001），且IL-9与GATA3和CRTH2共定位。在体外，CLL患者分选的ILC2s上清液中的IL-9水平升高。抗IL-9抗体治疗显著降低了PD-1 Treg和TIGIT Treg细胞，同时增加了颗粒酶B CD8 T细胞（P < 0.05）。然而，对Treg、CTLA-4 Treg和穿孔素CD8 T细胞无显著影响（P > 0.05）。此外，抗IL-9抗体显著增加了早期凋亡（P < 0.05）。
结论
ILC2s通过IL-9影响CD8 T细胞和Treg细胞，削弱CD8 T细胞的抗肿瘤作用，增强Treg细胞的免疫抑制作用，从而促进CLL发病机制。
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