Zeng Qingxiang, Xi Luo, Zeng Yinhui, Liu Wenlong, Zhou Lifeng
Department of Otolaryngology, Guangzhou Women and Children's Medical Center, Guangzhou Medical University, No. 9, Jinsui Road, Guangzhou, China.
World Allergy Organ J. 2022 Aug 10;15(8):100659. doi: 10.1016/j.waojou.2022.100659. eCollection 2022 Aug.
Osteopontin (OPN) can regulate Th2 inflammation in allergic rhinitis (AR). A recent study suggested that group II innate lymphoid cells (ILC2s) were very important for airway inflammation. But the role of OPN in ILC2s regulation is not explored.
Purified ILC2s were stimulated by human recombinant OPN. The expression of GATA3 and RORα was assayed using real-time polymerase chain reaction (PCR) and enzyme linked immunosorbent assay. MiR-181a was transfected into eosinophils to test the OPN production. The protein concentrations of interleukin (IL)-5 and IL-13 were examined using ELISA. Purified eosinophils and ILC2s were cocultured and stimulated by OPN and the activation of eosinophils was detected by ELISA.
After OPN stimulation, the ILC2s proliferation, the mRNA levels of GATA3 and RORα, the protein of GATA3, RORα, IL-5 and IL-13 expression were up-regulated significantly in a dose dependent manner. Eosinophils cultured alone transfected with miR-181a mimics produced less OPN protein compared with eosinophils transfected with miR-control, whereas OPN production was significantly promoted when miR-181a inhibitor was transfected. In the eosinophils and ILC2s coculture system, eosinophil cationic protein (ECP) production induced by OPN or IL-33 were significantly higher than ECP production in eosinophils culture system. OPN presented similar potency with IL-33 in the activation of eosinophils. When anti-IL-5 antibody was added, the production of ECP was significantly inhibited.
Our data for the first time provided new evidence that OPN played important roles in innate immunity of AR by regulation of ILC2s and the interaction between ILC2s and eosinophils.
骨桥蛋白(OPN)可调节变应性鼻炎(AR)中的Th2炎症。最近一项研究表明,II型固有淋巴细胞(ILC2s)对气道炎症非常重要。但OPN在ILC2s调节中的作用尚未得到探索。
用重组人OPN刺激纯化的ILC2s。采用实时聚合酶链反应(PCR)和酶联免疫吸附测定法检测GATA3和RORα的表达。将miR-181a转染至嗜酸性粒细胞中以检测OPN的产生。采用酶联免疫吸附测定法检测白细胞介素(IL)-5和IL-13的蛋白浓度。将纯化的嗜酸性粒细胞和ILC2s共培养,并用OPN刺激,通过酶联免疫吸附测定法检测嗜酸性粒细胞的活化情况。
OPN刺激后,ILC2s增殖、GATA3和RORα的mRNA水平、GATA3、RORα、IL-5和IL-13的蛋白表达均呈剂量依赖性显著上调。与转染miR对照的嗜酸性粒细胞相比,单独培养的转染miR-181a模拟物的嗜酸性粒细胞产生的OPN蛋白较少,而转染miR-181a抑制剂时OPN产生显著增加。在嗜酸性粒细胞和ILC2s共培养体系中,OPN或IL-33诱导的嗜酸性粒细胞阳离子蛋白(ECP)产生显著高于嗜酸性粒细胞培养体系中的ECP产生。OPN在激活嗜酸性粒细胞方面与IL-33具有相似的效力。加入抗IL-5抗体后,ECP的产生显著受到抑制。
我们的数据首次提供了新的证据,表明OPN通过调节ILC2s以及ILC2s与嗜酸性粒细胞之间的相互作用,在AR的固有免疫中发挥重要作用。