慢性鼻-鼻窦炎患者中表面活性蛋白D的表达及鼻外植体模型中体外对曲霉和链格孢的免疫反应
Surfactant protein d expression in chronic rhinosinusitis patients and immune responses in vitro to Aspergillus and alternaria in a nasal explant model.
作者信息
Ooi Eng Hooi, Wormald Peter-John, Carney A Simon, James Craig Lloyd, Tan Lor Wai
机构信息
Department of Surgery-Otorhinolaryngology Head and Neck Surgery, University of Adelaide and Flinders University, South Australia.
出版信息
Laryngoscope. 2007 Jan;117(1):51-7. doi: 10.1097/01.mlg.0000243196.75418.6f.
OBJECTIVES/HYPOTHESIS: Common fungi have been implicated in the pathogenesis of chronic rhinosinusitis (CRS) with eosinophilic mucus (EMCRS). Surfactant protein (SP)-D plays an important role in the immune response to Aspergillus fumigatus in the lungs. We sought to determine whether SP-D is expressed in nasal mucosa and investigated the response of SP-D in vitro to fungal allergens.
STUDY DESIGN AND METHODS
- Nasal biopsies from 59 CRS and EMCRS patients, stratified into allergic fungal sinusitis (AFS), nonallergic fungal eosinophilic sinusitis (NAFES), and nonallergic nonfungal eosinophilic sinusitis (NANFES) were studied by quantitative reverse-transcriptase polymerase chain reaction (qRT-PCR), immunostaining and enzyme-linked immunosorbent assay (ELISA). 2) Nasal tissue from three CRS and three NANFES patients was cultured with fungal allergens in a nasal explant in vitro model for 24 hours at increasing concentrations and mRNA SP-D secreted SP-D protein levels in response to the fungi determined by qRT-PCR and ELISA.
RESULTS
Staining for SP-D was detected in the submucosal glands from the nasal biopsies in all patient groups except for AFS. By ELISA, SP-D was undetectable in AFS and decreased in NAFES, NANFES, and CRS compared with controls. CRS patients in vitro cultured with Aspergillus and Alternaria allergens in a nasal tissue explant model induced up-regulation of SP-D by qRT-PCR. In contrast, NANFES nasal tissue explants cultured with Aspergillus allergens induced down-regulation of SP-D.
CONCLUSIONS
We report for the first time the expression of SP-D in both diseased and normal nasal mucosa. SP-D expression in CRS patients is up-regulated by fungal allergens in an in vitro model. These results may provide potential novel therapy for treatment of CRS.
目的/假设:常见真菌与伴有嗜酸性黏液的慢性鼻-鼻窦炎(CRS)的发病机制有关。表面活性蛋白(SP)-D在肺部对烟曲霉的免疫反应中起重要作用。我们试图确定SP-D是否在鼻黏膜中表达,并研究其在体外对真菌过敏原的反应。
研究设计与方法
1)对59例CRS和伴有嗜酸性黏液的CRS(EMCRS)患者的鼻活检组织进行研究,这些患者被分为变应性真菌性鼻窦炎(AFS)、非变应性真菌性嗜酸性鼻窦炎(NAFES)和非变应性非真菌性嗜酸性鼻窦炎(NANFES),采用定量逆转录聚合酶链反应(qRT-PCR)、免疫染色和酶联免疫吸附测定(ELISA)。2)将3例CRS患者和3例NANFES患者的鼻组织在体外鼻外植体模型中与真菌过敏原一起培养24小时,浓度逐渐增加,通过qRT-PCR和ELISA测定响应真菌时分泌的SP-D蛋白水平和mRNA SP-D水平。
结果
除AFS外,所有患者组鼻活检组织的黏膜下腺均检测到SP-D染色。通过ELISA检测,AFS中未检测到SP-D,与对照组相比,NAFES、NANFES和CRS中的SP-D减少。在鼻组织外植体模型中,用曲霉菌和链格孢菌过敏原体外培养CRS患者,通过qRT-PCR诱导SP-D上调。相反,用曲霉菌过敏原培养NANFES鼻组织外植体诱导SP-D下调。
结论
我们首次报道了SP-D在患病和正常鼻黏膜中的表达。在体外模型中,CRS患者的SP-D表达受真菌过敏原上调。这些结果可能为CRS的治疗提供潜在的新疗法。