Bergougnan Carolin, Dittlein Daniela C, Hümmer Elke, Riepl Rosalie, Eisenbart Selina, Böck Dominik, Griesbaum Lena, Weigl Anna, Damialis Athanasios, Hartwig Alexander, Neumann Avidan U, Zenk Johannes, Traidl-Hoffmann Claudia, Gilles Stefanie
Chair and Institute of Environmental Medicine, UNIKA-T, Technical University of Munich and HelmholtzZentrum München, Augsburg, Germany.
Christine-Kühne-Center for Allergy Research and Education (CK-Care), Davos, Switzerland.
World Allergy Organ J. 2020 Mar 9;13(3):100109. doi: 10.1016/j.waojou.2020.100109. eCollection 2020 Mar.
The epithelial cell-derived cytokine milieu has been discussed as a "master switch" in the development of allergic disease. To understand the role of innate immune response in nasal epithelial cells during allergic inflammation, we created and established a fast and minimally invasive method to isolate and culture human nasal epithelial cells from clinically and immunologically well characterized patients. Human nasal epithelial cells from non-atopic volunteers and from allergic rhinitis patients were compared in respect to their growth, barrier integrity, pattern recognition, receptor expression, and immune responses to allergens and an array of pathogen-associated molecular patterns and inflammasome activators. Cells from nasal scrapings were clearly identified as nasal epithelial cells by staining of pan-Cytokeratin, Cytokeratin-14 and Tubulin. Additionally, Mucin 5AC staining revealed the presence of goblet cells, while staining of tight-junction protein Claudin-1, Occludin and ZO-1 showed the ability of the cells to form a tight barrier. Cells of atopic donors grew slower than cells of non-atopic donors. All nasal epithelial cells expressed TLR1-6 and 9, yet the expression of TLR-9 was lower in cells from allergic rhinitis (AR) donors. Additionally, epithelial cells from AR donors responded with a different TLR expression pattern to stimulation with TLR ligands. TLR-3 was the most potent modulator of cytokine and chemokine secretion in all human nasal epithelial cells (HNECs). The secretion of IL-1β, CCL-5, IL-8, IL-18 and IL-33 was elevated in HNECs of AR donors as compared to cells of non-atopic donors. This was observed in the steady-state (IL-18, IL-33) as well as under stimulation with TLR ligands (IL-18, IL-33, CCL-5, IL-8), aqueous pollen extracts (IL-18, IL-33), or the inflammasome activator Nigericin (IL-1β). In conclusion, nasal epithelial cells of AR donors show altered physical barrier responses in steady-state and in response to allergen stimulation. Cells of AR donors show increased expression of pro-inflammatory and IL-1 family cytokines at baseline and under stimulation, which could contribute to a micromilieu which is favorable for Th2.
上皮细胞衍生的细胞因子环境被认为是过敏性疾病发展中的“主开关”。为了解天然免疫反应在变应性炎症期间鼻上皮细胞中的作用，我们创建并建立了一种快速且微创的方法，用于从临床和免疫学特征明确的患者中分离和培养人鼻上皮细胞。比较了非特应性志愿者和变应性鼻炎患者的人鼻上皮细胞在生长、屏障完整性、模式识别、受体表达以及对变应原和一系列病原体相关分子模式及炎性小体激活剂的免疫反应方面的差异。通过全细胞角蛋白、细胞角蛋白 -14 和微管蛋白染色，明确将鼻刮片中的细胞鉴定为鼻上皮细胞。此外，黏蛋白 5AC 染色显示存在杯状细胞，而紧密连接蛋白闭合蛋白 -1、闭合蛋白和闭锁小带 -1 的染色显示细胞具有形成紧密屏障的能力。特应性供体的细胞比非特应性供体的细胞生长得慢。所有鼻上皮细胞均表达 Toll 样受体（TLR）1 - 6 和 9，但变应性鼻炎（AR）供体细胞中 TLR - 9 的表达较低。此外，AR 供体的上皮细胞对 TLR 配体刺激的 TLR 表达模式不同。TLR - 3 是所有人类鼻上皮细胞（HNEC）中细胞因子和趋化因子分泌的最有效调节因子。与非特应性供体细胞相比，AR 供体的 HNEC 中白细胞介素 -1β（IL -1β）、趋化因子配体 5（CCL -5）、白细胞介素 -8、白细胞介素 -18 和白细胞介素 -33 的分泌升高。在稳态（IL -18、IL -33）以及 TLR 配体刺激（IL -18、IL -33、CCL -5、IL -8）、水性花粉提取物刺激（IL -18、IL -33）或炎性小体激活剂尼日利亚菌素刺激（IL -1β）下均观察到这种情况。总之，AR 供体的鼻上皮细胞在稳态和变应原刺激下显示出物理屏障反应改变。AR 供体的细胞在基线和刺激下促炎细胞因子和 IL -1 家族细胞因子的表达增加，这可能有助于形成有利于 Th2 的微环境。
