Hong Wenzhou, Khampang Pawjai, Samuels Tina L, Kerschner Joseph E, Yan Ke, Simpson Pippa
Department of Otolaryngology and Communication Sciences, Medical College of Wisconsin, Milwaukee, WI 53226, USA.
Department of Otolaryngology and Communication Sciences, Medical College of Wisconsin, Milwaukee, WI 53226, USA.
Int J Pediatr Otorhinolaryngol. 2017 Oct;101:30-36. doi: 10.1016/j.ijporl.2017.07.025. Epub 2017 Jul 21.
Calgranulins (calcium-binding proteins S100A8, S100A9 and S100A12) are predominant cytoplasmic proteins of neutrophils and produced by various cells, playing multiple functions in innate immunity and the inflammatory process. Although up-regulated expression of S100A8 and S100A9 genes were observed in an animal model of otitis media (OM), their expressions have not been studied in human middle ear epithelial cells in response to the OM pathogen or in patients with recurrent or chronic OM (recurrent OM/RecOM or chronic OM with effusion/COME).
Gene expressions were compared between Streptococcus pneumoniae (SP)-infected and non-infected human middle ear epithelial cells (HMEECs) as well as between chronic OM patients and control patients (CI).
Gene expressions were profiled by quantitative real time PCR (qPCR). S100 proteins in OM patient and CI middle ear biopsies were detected by immunostaining.
S100A8, S100A9 and S100A12 gene expressions were elevated in SP-infected HMEECs in time-dependent manner. S100A8 and S100A9 but not S100A12 gene expression was significantly elevated in the middle ear mucosa of OM patients. S100A8 and S100A9 protein were observed in middle ear mucosa of OM, but not CI patients. Minimal co-localization was observed between S100A8 and S100A9 with neutrophil elastase and cytokeratin in ME sections of OM patients.
Elevated S100A8 and S100A9 gene expression in SP-infected HMEECs and in the middle ear mucosa of OM, minor co-localized with neutrophil markers suggests that middle ear epithelial cell secretion of S100A8 and S100A9 may play a role in the pathogenesis of recurrent and chronic OM.
钙粒蛋白(钙结合蛋白S100A8、S100A9和S100A12)是中性粒细胞的主要细胞质蛋白,由多种细胞产生,在先天免疫和炎症过程中发挥多种功能。尽管在中耳炎(OM)动物模型中观察到S100A8和S100A9基因表达上调,但尚未研究它们在人类中耳上皮细胞中对OM病原体的反应,或在复发性或慢性OM(复发性OM/RecOM或慢性渗出性中耳炎/COME)患者中的表达情况。
比较肺炎链球菌(SP)感染和未感染的人类中耳上皮细胞(HMEECs)之间以及慢性OM患者和对照患者(CI)之间的基因表达。
通过定量实时PCR(qPCR)分析基因表达。通过免疫染色检测OM患者和CI中耳活检组织中的S100蛋白。
SP感染的HMEECs中,S100A8、S100A9和S100A12基因表达呈时间依赖性升高。OM患者中耳黏膜中S100A8和S100A9基因表达显著升高,但S100A12基因表达未升高。在OM患者的中耳切片中,OM患者的中耳黏膜中观察到S100A8和S100A9蛋白,但CI患者未观察到。在OM患者的中耳切片中,S100A8和S100A9与中性粒细胞弹性蛋白酶和细胞角蛋白的共定位极少。
SP感染的HMEECs和OM中耳黏膜中S100A8和S100A9基因表达升高,与中性粒细胞标志物的共定位较少,提示中耳上皮细胞分泌S100A8和S100A9可能在复发性和慢性OM的发病机制中起作用。