Chen Yifang, Li Yang, Guo Han, Zhang Zhaoqi, Zhang Jiayu, Dong Xue, Liu Yi, Zhuang Yuan, Zhao Yong
Department of State Key Laboratory of Membrane Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, People's Republic of China.
Savaid Medical School, University of Chinese Academy of Sciences, Beijing, People's Republic of China.
J Inflamm Res. 2021 Sep 16;14:4669-4686. doi: 10.2147/JIR.S329528. eCollection 2021.
Neutrophils present great diverse phenotypes in various microenvironments and play different immune regulatory functions. Neutrophils generally classified into inflammatory phenotype N1 and anti-informatory phenotype N2. Our recent studies showed that IL-23 alone stimulated neutrophils to express IL-17A, IL-17F and IL-22 and displayed a gene transcriptional profile similar to Th17 cells. In the present study, we tried to identify potential cytokines to promote IL-23-induced neutrophil polarization.
Mouse bone marrow-derived neutrophils and human peripheral blood neutrophils were treated with IL-23 (10 ng/mL) plus IL-18 (25 ng/mL) to induce Th17-like subset in vitro and detected by real-time PCR, flow cytometry, ELISA, immunofluorescence and RNA-seq assays. In vivo, collagen-induced arthritis (CIA) mouse model and EL4 tumor-bearing mouse model were used to characterize the potential roles of N(IL-23+IL-18) in inflammation and tumor.
Real-time PCR, ELISA and flow cytometry assays showed that IL-18 could significantly enhance IL-23-induced IL-17A, IL-17F and IL-22 expressions in mouse and human neutrophils in a synergistic way, although IL-18 alone failed to induce these cytokines expression. RNA-seq and molecular studies showed that the polarization of N(IL-23+IL-18) is mainly mediated by the JNK/p38-STAT3-BATF signaling pathway. Adoptive transfer of the induced N(IL-23+IL-18) neutrophils significantly accelerated the tumor growth in EL4 tumor-bearing mice and enhanced disease progression in the CIA mouse model. IL-17A-deficient N(IL-23+IL-18) neutrophils failed to enhance the CIA pathogenesis in this model, suggesting that IL-17A may be involved in the N(IL-23+IL-18) neutrophils-promoted arthritis in mice.
The Th17-type subpopulation N(IL-23+IL-18) has pro-tumor and pro-inflammatory properties. Recognizing the different functional polarization of neutrophils would significantly help us to understand the distinctive protective/pathological roles of neutrophils in physiological and different pathological situations.
中性粒细胞在各种微环境中呈现出多种不同的表型，并发挥不同的免疫调节功能。中性粒细胞通常分为炎症表型N1和抗炎表型N2。我们最近的研究表明，单独的白细胞介素-23（IL-23）刺激中性粒细胞表达白细胞介素-17A（IL-17A）、白细胞介素-17F（IL-17F）和白细胞介素-22（IL-22），并呈现出与辅助性T细胞17（Th17）细胞相似的基因转录谱。在本研究中，我们试图确定促进IL-23诱导的中性粒细胞极化的潜在细胞因子。
用IL-23（10纳克/毫升）加IL-18（25纳克/毫升）处理小鼠骨髓来源的中性粒细胞和人外周血中性粒细胞，以在体外诱导Th17样亚群，并通过实时聚合酶链反应（PCR）、流式细胞术、酶联免疫吸附测定（ELISA）、免疫荧光和RNA测序（RNA-seq）分析进行检测。在体内，采用胶原诱导的关节炎（CIA）小鼠模型和EL4荷瘤小鼠模型来表征N（IL-23+IL-18）在炎症和肿瘤中的潜在作用。
实时PCR、ELISA和流式细胞术分析表明，IL-18可协同显著增强IL-23诱导的小鼠和人中性粒细胞中IL-17A、IL-17F和IL-22的表达，尽管单独的IL-18未能诱导这些细胞因子的表达。RNA-seq和分子研究表明，N（IL-23+IL-18）的极化主要由应激活化蛋白激酶（JNK）/p38-信号转导子和转录激活子3（STAT3）-BATF信号通路介导。过继转移诱导的N（IL-23+IL-18）中性粒细胞显著加速了EL4荷瘤小鼠的肿瘤生长，并增强了CIA小鼠模型中的疾病进展。在该模型中，缺乏IL-17A的N（IL-23+IL-18）中性粒细胞未能增强CIA的发病机制，这表明IL-17A可能参与了N（IL-23+IL-18）中性粒细胞促进的小鼠关节炎。
Th17型亚群N（IL-23+IL-18）具有促肿瘤和促炎特性。认识到中性粒细胞不同的功能极化将极大地帮助我们理解中性粒细胞在生理和不同病理情况下独特的保护/病理作用。
