Wang Chen, Zhuo Jin-Jing, Li Wen-Qian, Zhou Min-Li, Cheng Ke-Jia
Department of Otolaryngology, The First Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, China.
World Allergy Organ J. 2024 Jan 17;17(2):100852. doi: 10.1016/j.waojou.2023.100852. eCollection 2024 Feb.
Roles of ILC2s in allergic rhinitis (AR) and local allergic rhinitis (LAR) are unclear. In this study, we are determined to find the levels of autophagy and mitophagy of ILC2s in allergic nasal inflammation.
ELISA was used to detect type 2 inflammatory cytokines. Hematoxylin and eosin (H&E) staining were used to compare the eosinophil (EOS) infiltration of nasal tissue specimens. Flow cytometry was used to detect the levels of ILC2s and Th2 cells. Immunohistochemistry (IHC) and Western blot (WB) were used to detect the levels of Beclin1, LC3, p62, PINK1, Parkin, FUNDC1, and BNIP3 in nasal mucosa. The levels of autophagy related proteins and mitophagy related proteins of the ILC2s were detected by WB. The number of autophagosomes of ILC2s was observed by transmission electron microscopy. The co-localization levels of GFP-LC3 and Mito tracker in ILC2s were observed by confocal microscopy using immunofluorescence.
We found that the level of type 2 inflammation in AR and LAR mice was significantly increased. The levels of autophagy and mitophagy of AR and LAR mice in nasal mucosa and ILC2s were both increased.
ILC2s may be associated with the occurrence and development of nasal allergic inflammation. The abnormal increase of autophagy and mitophagy levels in the nose may be associated with the incidence of AR and LAR. Abnormal autophagy and mitophagy levels of ILC2s cells may be one of the causes of allergic nasal inflammation.
2型固有淋巴细胞(ILC2s)在变应性鼻炎(AR)和局部变应性鼻炎(LAR)中的作用尚不清楚。在本研究中,我们旨在探究变应性鼻炎症中ILC2s的自噬和线粒体自噬水平。
采用酶联免疫吸附测定(ELISA)检测2型炎性细胞因子。采用苏木精-伊红(H&E)染色比较鼻组织标本中嗜酸性粒细胞(EOS)浸润情况。采用流式细胞术检测ILC2s和Th2细胞水平。采用免疫组织化学(IHC)和蛋白质免疫印迹法(WB)检测鼻黏膜中Beclin1、微管相关蛋白1轻链3(LC3)、p62、无翅型MMTV整合位点家族成员1(PINK1)、帕金蛋白、含 FUN14 结构域蛋白1(FUNDC1)和BCL2/腺病毒E1B 19kDa相互作用蛋白3(BNIP3)水平。通过WB检测ILC2s的自噬相关蛋白和线粒体自噬相关蛋白水平。通过透射电子显微镜观察ILC2s的自噬体数量。采用免疫荧光共聚焦显微镜观察绿色荧光蛋白标记的LC3(GFP-LC3)与线粒体示踪剂在ILC2s中的共定位水平。
我们发现AR和LAR小鼠的2型炎症水平显著升高。AR和LAR小鼠鼻黏膜和ILC2s中的自噬和线粒体自噬水平均升高。
ILC2s可能与鼻变应性炎症的发生和发展有关。鼻腔中自噬和线粒体自噬水平的异常升高可能与AR和LAR的发病有关。ILC2s细胞自噬和线粒体自噬水平异常可能是变应性鼻炎症的原因之一。