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.

Abstract

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

  1. 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.

摘要

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。

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