Martens Katleen, De Boeck Ilke, Jokicevic Katarina, Kiekens Filip, Farré Ricard, Vanderveken Olivier M, Seys Sven F, Lebeer Sarah, Hellings Peter W, Steelant Brecht
KU Leuven, Department of Microbiology, Immunology and Transplantation, Allergy and Clinical Immunology Research Group, Leuven, Belgium.
Department of Bioscience Engineering, Research Group Environmental Ecology and Applied Microbiology, University of Antwerp, Antwerp, Belgium.
Allergy Asthma Immunol Res. 2021 Jul;13(4):560-575. doi: 10.4168/aair.2021.13.4.560.
A defective epithelial barrier has been demonstrated in chronic rhinosinusitis with nasal polyps (CRSwNP). Lactobacilli are shown to restore epithelial barrier defects in gastrointestinal disorders, but their effect on the airway epithelial barrier is unknown. In this study, hence, we evaluated whether the nasopharyngeal isolates AMBR2 and AMBR8 could restore nasal epithelial barrier integrity in CRSwNP.
trans-epithelial tissue resistance and fluorescein isothiocyanate-dextran 4 kDa (FD4) permeability of nasal mucosal explants were measured. The relative abundance of lactobacilli in the maxillary sinus of CRSwNP patients was analyzed by amplicon sequencing of the V4 region of the 16S rRNA gene. The effect of spray-dried AMBR2 and AMBR8 on epithelial integrity was investigated in primary nasal epithelial cells (pNECs) from healthy controls and patients with CRSwNP as well as in a murine model of interleukin (IL)-4 induced barrier dysfunction. The activation of Toll-like receptor 2 (TLR2) was explored by using polyclonal antibodies.
Patients with CRSwNP had a defective epithelial barrier which positively correlated with the relative abundance of lactobacilli-specific amplicons in the maxillary sinus. AMBR2, but not AMBR8, increased the trans-epithelial electrical resistance (TEER) of pNECs from CRSwNP patients in a time-dependent manner. Treatment of epithelial cells with AMBR2 promoted the tight junction proteins occludin and zonula occludens-1 reorganization. Furthermore, AMBR2 prevented IL-4-induced nasal permeability and . Finally, the beneficial effect of AMBR2 on nasal epithelial cells was TLR2-dependent as blocking TLR2 receptors prevented the increase in TEER.
A defective epithelial barrier in CRSwNP may be associated with a decrease in relative abundance of lactobacilli-specific amplicons. AMBR2 would restore nasal epithelial integrity and can be a novel therapeutic strategy for CRSwNP.
在伴有鼻息肉的慢性鼻-鼻窦炎（CRSwNP）中已证实存在上皮屏障缺陷。乳酸菌已被证明可修复胃肠道疾病中的上皮屏障缺陷，但其对气道上皮屏障的影响尚不清楚。因此，在本研究中，我们评估了鼻咽分离株AMBR2和AMBR8是否能恢复CRSwNP中的鼻上皮屏障完整性。
测量鼻黏膜外植体的跨上皮组织电阻和4 kDa异硫氰酸荧光素-葡聚糖（FD4）通透性。通过对16S rRNA基因V4区域进行扩增子测序，分析CRSwNP患者上颌窦中乳酸菌的相对丰度。研究喷雾干燥的AMBR2和AMBR8对来自健康对照和CRSwNP患者的原代鼻上皮细胞（pNECs）以及白细胞介素（IL）-4诱导的屏障功能障碍小鼠模型上皮完整性的影响。使用多克隆抗体探索Toll样受体2（TLR2）的激活情况。
CRSwNP患者存在上皮屏障缺陷，这与上颌窦中乳酸菌特异性扩增子的相对丰度呈正相关。AMBR2而非AMBR8能以时间依赖性方式增加CRSwNP患者pNECs的跨上皮电阻（TEER）。用AMBR2处理上皮细胞可促进紧密连接蛋白闭合蛋白和闭合小带蛋白-1的重组。此外，AMBR2可预防IL-4诱导的鼻通透性增加。最后，AMBR2对鼻上皮细胞的有益作用依赖于TLR2，因为阻断TLR2受体会阻止TEER的增加。
CRSwNP中的上皮屏障缺陷可能与乳酸菌特异性扩增子的相对丰度降低有关。AMBR2可恢复鼻上皮完整性，可能成为CRSwNP的一种新的治疗策略。
