Epithelial, connective tissue and immune cells contribute in various ways to the pathophysiology of chronic rhinosinusitis (CRS). However, data of their distribution in upper airway mucosa are sparse. We aimed to provide quantitative, purely informative data on the distribution of these cell lineages and their coexpression patterns, which might help identifying, e.g., cells in the epithelium undergoing through epithelial–mesenchymal transition (EMT). For this purpose, we used immunofluorescence multichannel image cytometry (IMIC). We examined fixed paraffin-embedded tissue samples (FFPE) of six patients with chronic rhinosinusitis (CRS) and of three patients without CRS (controls). The direct-conjugated antibodies pancytokeratin, vimentin and CD45/CD18 were used for coexpression analysis in epithelial layer and lamina propria. Image acquisition and analysis were performed with TissueFAXS and StrataQuest, respectively. To distinguish positive from negative expression, a ratio between cell-specific immunostaining intensity and background was developed. Isotype controls were used as negative controls. Per patient, a 4.5-mm² tissue area was scanned and a median of 14,875 cells was recognized. The most common cell types were cytokeratin-single-positive (26%), vimentin-single-positive (13%) and CD45/CD18-single-positive with CD45/CD18–vimentin-double-positive cells (29%). In the patients with CRS, CD45/CD18-single-positive cells were 3–6 times higher compared to the control patients. In the epithelial layer, cytokeratin–vimentin-double-positive EMT cells were observed 3–5 times higher in the patients with CRS than in the control patients. This study provided quantitative data for the distribution of crucial cell types in CRS. Future studies may focus on the distribution and coexpression patterns of different immune cells in CRS or even cancer tissue.

上皮细胞、结缔组织和免疫细胞以多种方式影响慢性鼻窦炎 (CRS) 的病理生理学。然而，它们在上呼吸道粘膜中分布的数据很少。我们的目的是提供有关这些细胞谱系的分布及其共表达模式的定量、纯信息性数据，这可能有助于识别例如经历上皮-间质转化（EMT）的上皮细胞。为此，我们使用了免疫荧光多通道图像细胞术（IMIC）。我们检查了 6 名慢性鼻窦炎 (CRS) 患者和 3 名无 CRS 患者（对照）的固定石蜡包埋组织样本 (FFPE)。使用直接缀合抗体全细胞角蛋白、波形蛋白和 CD45/CD18 进行上皮层和固有层的共表达分析。分别使用 TissueFAXS 和 StrataQuest 进行图像采集和分析。为了区分阳性表达和阴性表达，制定了细胞特异性免疫染色强度和背景之间的比率。使用同型对照作为阴性对照。对每位患者扫描 4.5 mm ^{2 的}组织区域，识别出中位数为 14,875 个细胞。最常见的细胞类型是细胞角蛋白单阳性（26%）、波形蛋白单阳性（13%）和CD45/CD18单阳性以及CD45/CD18-波形蛋白双阳性细胞（29%）。在 CRS 患者中，CD45/CD18 单阳性细胞比对照患者高 3-6 倍。在上皮层，CRS 患者的细胞角蛋白-波形蛋白双阳性 EMT 细胞比对照患者高 3-5 倍。这项研究为 CRS 中关键细胞类型的分布提供了定量数据。 未来的研究可能集中于不同免疫细胞在 CRS 甚至癌组织中的分布和共表达模式。