Szopa Iwona Monika, Majchrzak-Kuligowska Kinga, Pingwara Rafał, Kulka Marek, Taşdemir Monika, Gajewska Małgorzata
Department of Physiological Sciences, Institute of Veterinary Medicine, Warsaw University of Life Sciences, 02-776 Warsaw, Poland.
Department of Pathology and Veterinary Diagnostics, Institute of Veterinary Medicine, Warsaw University of Life Sciences, 02-776 Warsaw, Poland.
Int J Mol Sci. 2025 May 21;26(10):4946. doi: 10.3390/ijms26104946.
Th17 lymphocytes are a distinct subpopulation of T cells that are characterized by the production of interleukins IL-17, IL-21, IL-22, and IL-26, and high expression of RORγt. These cells play an important role in inflammation and autoimmune diseases. Recent studies using rodent and human models have also highlighted their promising properties as agents in cellular immunotherapy for cancer. However, much less is known about the properties of canine Th17 lymphocytes, despite the domestic dog being an important model used in comparative medicine. In this study, we developed methods of activation and differentiation of canine CD4 T lymphocytes towards the Th17 phenotype. Additionally, we targeted the Wnt/β-catenin signaling pathway to modulate the efficiency of Th17 cells differentiation. CD4 T cells were successfully activated with magnetic EpoxyBeads, and in combination with the appropriate programming medium, they acquired the Th17 phenotype. Furthermore, indomethacin, an inhibitor of the Wnt/β-catenin pathway, significantly increased the efficiency of differentiation, causing elevated production of IL-17 and changed T cell metabolism by promoting oxidative phosphorylation. The protocol elaborated in our study provides an efficient method of canine Th17 lymphocyte differentiation. Our findings also suggested that the modification of the Wnt/β-catenin signaling pathway could be a valuable strategy for optimizing canine Th17 cell differentiation and advancing cell-based immunotherapy.
辅助性T细胞17（Th17）淋巴细胞是T细胞的一个独特亚群，其特征是产生白细胞介素IL-17、IL-21、IL-22和IL-26，以及维甲酸相关孤儿受体γt（RORγt）的高表达。这些细胞在炎症和自身免疫性疾病中起重要作用。最近使用啮齿动物和人类模型的研究也突出了它们作为癌症细胞免疫治疗药物的潜在特性。然而，尽管家犬是比较医学中使用的重要模型，但对犬Th17淋巴细胞的特性了解却少得多。在本研究中，我们开发了将犬CD4 T淋巴细胞激活并分化为Th17表型的方法。此外，我们靶向Wnt/β-连环蛋白信号通路来调节Th17细胞分化的效率。用磁性环氧珠成功激活了CD4 T细胞，并与适当的编程培养基结合，它们获得了Th17表型。此外，Wnt/β-连环蛋白途径的抑制剂吲哚美辛显著提高了分化效率，导致IL-17产生增加，并通过促进氧化磷酸化改变了T细胞代谢。我们研究中详细阐述的方案提供了一种有效的犬Th17淋巴细胞分化方法。我们的研究结果还表明，修饰Wnt/β-连环蛋白信号通路可能是优化犬Th17细胞分化和推进基于细胞的免疫治疗的一种有价值的策略。
