Kohanski Michael A, Lane Andrew P
Department of Otolaryngology-Head and Neck Surgery, The Johns Hopkins University School of Medicine, Baltimore, Maryland.
JAMA Otolaryngol Head Neck Surg. 2015 Apr;141(4):341-9. doi: 10.1001/jamaoto.2014.3550.
Chronic rhinosinusitis (CRS) is an inflammatory disorder of the nose and paranasal sinuses. Staphylococcus aureus is increasingly linked with CRS exacerbations. Little is known about how bacteria activate inflammatory pathways that contribute to CRS.
To develop an in vitro coculture system to explore how infection with S aureus stimulates innate immune responses of sinonasal epithelial cells (SNECs).
DESIGN, SETTING, AND PARTICIPANTS: Sinonasal epithelial cells were collected from 13 patients during endoscopic sinus surgery and grown in culture at the air-liquid interface from July 2014 through December 2014.
Differentiated SNECs from control individuals, patients with CRS with nasal polyps (CRSwNPs), and patients with CRS without nasal polyps (CRSsNPs) were infected with S aureus at 3 different concentrations for 24 hours.
Growth of S aureus and viability of SNECs were measured. Expression of inflammatory markers and innate immune genes was measured by reverse transcription-polymerase chain reaction. Basal secretion of interleukin 8 was determined by enzyme-linked immunosorbent assay.
Cultured SNECs from patients with CRSsNPs demonstrated a significant increase (P < .05) in expression of interleukin 8 (23-fold to 82-fold) and tumor necrosis factor (11-fold to 61-fold) at all the tested concentrations of S aureus. Control or CRSwNP SNECs demonstrated a significant increase (P < .05) in expression of interleukin 8 (47-fold and 50-fold, respectively) and tumor necrosis factor (106-fold and 58-fold, respectively) at the higher inoculum of S aureus. Basal secretion of inflammatory markers correlated with expression changes. No significant changes in expression were observed for the helper T cell, subtype 2, inflammatory mediators tested.
In this study, we developed a model to study early innate immune-mediated changes in SNECs cocultured at an air-liquid interface with bacteria. We also demonstrated that bacterial burden can be detected by SNECs in the absence of adaptive immune-mediated responses. The CRSsNP SNECs are more sensitive to S aureus burden than control or CRSwNP SNECs. Future studies will further develop this infection model and explore the SNEC innate immune response to bacteria.
慢性鼻-鼻窦炎（CRS）是一种鼻和鼻窦的炎症性疾病。金黄色葡萄球菌与CRS病情加重的关联日益增加。关于细菌如何激活导致CRS的炎症途径，人们了解甚少。
建立一种体外共培养系统，以探究金黄色葡萄球菌感染如何刺激鼻窦上皮细胞（SNECs）的固有免疫反应。
设计、场所和参与者：2014年7月至12月期间，在鼻内镜鼻窦手术中从13名患者收集鼻窦上皮细胞，并在气液界面进行培养。
将来自对照个体、伴有鼻息肉的CRS患者（CRSwNP）和不伴有鼻息肉的CRS患者（CRSsNP）的分化SNECs分别用3种不同浓度的金黄色葡萄球菌感染24小时。
测量金黄色葡萄球菌的生长情况和SNECs的活力。通过逆转录-聚合酶链反应测量炎症标志物和固有免疫基因的表达。通过酶联免疫吸附测定法测定白细胞介素8的基础分泌量。
在所有测试的金黄色葡萄球菌浓度下，来自CRSsNP患者的培养SNECs的白细胞介素8表达显著增加（P < 0.05）（23倍至82倍）和肿瘤坏死因子表达显著增加（11倍至61倍）。在较高接种量的金黄色葡萄球菌作用下，对照或CRSwNP的SNECs的白细胞介素8表达分别显著增加（P < 0.05）（分别为47倍和50倍）和肿瘤坏死因子表达分别显著增加（分别为106倍和58倍）。炎症标志物的基础分泌量与表达变化相关。对于所测试的辅助性T细胞2型炎症介质，未观察到表达有显著变化。
在本研究中，我们建立了一个模型来研究在气液界面与细菌共培养的SNECs中早期固有免疫介导的变化。我们还证明，在缺乏适应性免疫介导反应的情况下，SNECs能够检测到细菌负荷。CRSsNP的SNECs对金黄色葡萄球菌负荷比对照或CRSwNP的SNECs更敏感。未来的研究将进一步完善这种感染模型，并探索SNECs对细菌的固有免疫反应。
