P2RX1阻断的中性粒细胞诱导CD8 T细胞功能障碍并影响胃癌细胞的免疫逃逸。

P2RX1-blocked neutrophils induce CD8 T cell dysfunction and affect the immune escape of gastric cancer cells.

作者信息

Zhang Yan, Zhang Fenglin, Liu Zhi, Li Min, Wu Ge, Li Hui

机构信息

Department of Medical Oncology, Ma'anshan People's Hospital, 519 Hunan East Road, Huashan District, Ma'anshan City, Anhui Province 243000, China.

出版信息

Cell Immunol. 2025 Feb;408:104901. doi: 10.1016/j.cellimm.2024.104901. Epub 2024 Dec 4.

DOI:10.1016/j.cellimm.2024.104901

PMID:39675308

Abstract

BACKGROUND

Gastric cancer (GC) is one of the deadly malignancies of the gastrointestinal tract. Research has confirmed the linkage of P2RX1 with immune cell activation and tumor progression. This project focused on the impact of P2RX1 level in neutrophils on the efficacy of immune checkpoint inhibitor (ICI) treatment in GC.

METHODS

Blood samples from 23 GC patients eligible for camrelizumab treatment were collected. Flow cytometry was carried out to analyze the proportion of P2RX1 in neutrophils. IHC was utilized to detect the expression level of PD-L1. We also evaluated the chemotaxis ability of neutrophils using a Transwell system, assessed the viability and apoptosis rate of GC cells using CCK-8 and flow cytometry, measured the proportions of CD8PD-1 and CD8GZMB cells, determined the expression levels of IL-6, TNFα, IFN-γ, IL-8, IL-12, IL-1β, and GZMB by utilizing enzyme-linked immunosorbent assay (ELISA), and examined the expression levels of P2RX1 and PD-L1 using western blot (WB). By establishing a xenograft mouse model, we studied the impact of P2RX1-blocked neutrophils on the efficacy of ICI treatment in the GC microenvironment.

RESULTS

In GC, clinical analysis revealed increased infiltration of P2RX1-lowly expressed neutrophil subsets and increased expression of PD-L1. In vitro experiments demonstrated that abnormal expression of P2RX1 affected neutrophil function. Furthermore, the blockage or knockdown of P2RX1 in neutrophils modulated CD8 T cell function, promoting GC progression. In in vivo experiments, the blockage of P2RX1 in neutrophils inhibited the effectiveness of ICI treatment in the GC microenvironment.

CONCLUSION

This project validated that the loss of P2RX1 in neutrophils induces CD8 T cell dysfunction and affects the GC development, indicating that P2RX1 may be an accurate biomarker for predicting ICI response, thus providing a theoretical basis for the clinical application of ICI.

摘要

背景

胃癌（GC）是胃肠道致命的恶性肿瘤之一。研究已证实P2RX1与免疫细胞激活及肿瘤进展之间存在联系。本项目聚焦于中性粒细胞中P2RX1水平对GC免疫检查点抑制剂（ICI）治疗疗效的影响。

方法

收集了23例符合卡瑞利珠单抗治疗条件的GC患者的血液样本。采用流式细胞术分析中性粒细胞中P2RX1的比例。利用免疫组化检测PD-L1的表达水平。我们还使用Transwell系统评估中性粒细胞的趋化能力，使用CCK-8和流式细胞术评估GC细胞的活力和凋亡率，测定CD8⁺PD-1和CD8⁺GZMB细胞的比例，利用酶联免疫吸附测定（ELISA）测定IL-6、TNFα、IFN-γ、IL-8、IL-12、IL-1β和GZMB的表达水平，并使用蛋白质免疫印迹（WB）检测P2RX1和PD-L1的表达水平。通过建立异种移植小鼠模型，我们研究了P2RX1阻断的中性粒细胞对GC微环境中ICI治疗疗效的影响。

结果

在GC中，临床分析显示P2RX1低表达中性粒细胞亚群的浸润增加以及PD-L1的表达增加。体外实验表明P2RX1的异常表达影响中性粒细胞功能。此外，中性粒细胞中P2RX1的阻断或敲低调节了CD8⁺T细胞功能，促进了GC进展。在体内实验中，中性粒细胞中P2RX1的阻断抑制了GC微环境中ICI治疗的有效性。