Matsumoto Haruki, Sudo Ryota, Fujita Yuya, Onizawa Michio, Saito Kenji, Sumichika Yuya, Yoshida Shuhei, Temmoku Jumpei, Matsuoka Naoki, Asano Tomoyuki, Sato Shuzo, Suzuki Eiji, Machida Takeshi, Migita Kiyoshi
Department of Rheumatology, Fukushima Medical University School of Medicine, 1 Hikarigaoka, Fukushima, Fukushima, 960-1295, Japan.
Department of Gastroenterology and Hepatology, Fukushima Medical University School of Medicine, 1 Hikarigaoka, Fukushima, Fukushima, 960-1295, Japan.
BMC Immunol. 2024 Oct 1;25(1):63. doi: 10.1186/s12865-024-00656-6.
Carcinoembryonic-antigen-related cell-adhesion molecule 1 (CEACAM1) is an adhesion molecule that acts as a coinhibitory receptor in the immune system. We previously demonstrated that CEACAM1 is predominantly expressed on peripheral blood neutrophils in patients with RA. The aim of the present study was to investigate the effects of Janus kinase inhibitors (JAKi) on cytokine-activated human neutrophils and CEACAM1 expression.
Peripheral blood neutrophils were obtained from healthy subjects. Isolated neutrophils were stimulated with tumor necrosis factor-alpha (TNF-α) or granulocyte-macrophage colony-stimulating factor (GM-CSF) in the presence or absence of JAKi. The expression of CEACAM1 in peripheral blood neutrophils was analyzed by flow cytometry. Protein phosphorylation of signal transducer and activator of transcription (STAT)1, STAT3, and STAT5 was assessed by western blot using phospho-specific antibodies.
We found that TNF-α-induced CEACAM1 expression was marginally suppressed after pretreatment with pan-JAK inhibitor, tofacitinib. Moreover, TNF-α induced STAT1 and STAT3 phosphorylation at the late stimulation phase (4 to 16 h). The expressions of CEACAM1 on neutrophils were markedly up-regulated by GM-CSF not by interleukin (IL)-6 stimulation. All JAKi inhibited GM-CSF-induced CEACAM1 expressions on neutrophils, however, the inhibitory effects of baricitinib were larger compared to those of tofacitinib or filgotinib. Moreover, CEACAM1 was marginally upregulated in interferon (IFN)-γ stimulated neutrophils. Similarly, JAKi inhibited IFN-γ-induced CEACAM1 expressions on neutrophils.
We demonstrated that JAKi prevent GM-CSF-induced CEACAM1 expression in neutrophils, and JAKi-induced inhibition depends on their selectivity against JAK isoforms. These findings suggest that JAKi can modulate the expression of CEACAM1 in cytokine-activated neutrophils, thereby limiting their activation.
癌胚抗原相关细胞黏附分子1（CEACAM1）是一种黏附分子，在免疫系统中作为共抑制受体发挥作用。我们之前证明，CEACAM1在类风湿关节炎（RA）患者的外周血中性粒细胞上主要表达。本研究的目的是探讨Janus激酶抑制剂（JAKi）对细胞因子激活的人中性粒细胞和CEACAM1表达的影响。
从健康受试者获取外周血中性粒细胞。在存在或不存在JAKi的情况下，用肿瘤坏死因子-α（TNF-α）或粒细胞-巨噬细胞集落刺激因子（GM-CSF）刺激分离的中性粒细胞。通过流式细胞术分析外周血中性粒细胞中CEACAM1的表达。使用磷酸化特异性抗体通过蛋白质印迹法评估信号转导和转录激活因子（STAT）1、STAT3和STAT5的蛋白质磷酸化。
我们发现，用泛JAK抑制剂托法替布预处理后，TNF-α诱导的CEACAM1表达略有抑制。此外，TNF-α在刺激后期（4至16小时）诱导STAT1和STAT3磷酸化。GM-CSF而非白细胞介素（IL）-6刺激显著上调中性粒细胞上CEACAM1的表达。所有JAKi均抑制GM-CSF诱导的中性粒细胞上CEACAM1的表达，然而，巴瑞替尼的抑制作用比托法替布或非戈替尼更大。此外，CEACAM1在干扰素（IFN）-γ刺激的中性粒细胞中略有上调。同样，JAKi抑制IFN-γ诱导的中性粒细胞上CEACAM1的表达。
我们证明JAKi可预防GM-CSF诱导的中性粒细胞中CEACAM1的表达，且JAKi诱导的抑制作用取决于它们对JAK异构体的选择性。这些发现表明，JAKi可调节细胞因子激活的中性粒细胞中CEACAM1的表达，从而限制其激活。
