Łaniewski Paweł, Gomez Adriana, Hire Geoffrey, So Magdalene, Herbst-Kralovetz Melissa M
Department of Basic Medical Sciences, College of Medicine-Phoenix, University of Arizona, Phoenix, Arizona, USA.
Department of Immunobiology and BIO5 Institute, College of Medicine-Tucson, University of Arizona, Tucson, Arizona, USA.
Infect Immun. 2017 Feb 23;85(3). doi: 10.1128/IAI.01049-16. Print 2017 Mar.
Colonization of the endometrium by pathogenic bacteria ascending from the lower female reproductive tract (FRT) is associated with many gynecologic and obstetric health complications. To study these host-microbe interactions , we developed a human three-dimensional (3-D) endometrial epithelial cell (EEC) model using the HEC-1A cell line and the rotating wall vessel (RWV) bioreactor technology. Our model, composed of 3-D EEC aggregates, recapitulates several functional/structural characteristics of human endometrial epithelial tissue, including cell differentiation, the presence of junctional complexes/desmosomes and microvilli, and the production of membrane-associated mucins and Toll-like receptors (TLRs). TLR function was evaluated by exposing the EEC aggregates to viral and bacterial products. Treatment with poly(I·C) and flagellin but not with synthetic lipoprotein (fibroblast-stimulating lipoprotein 1 [FSL-1]) or lipopolysaccharide (LPS) significantly induced proinflammatory mediators in a dose-dependent manner. To simulate ascending infection, we infected EEC aggregates with commensal and pathogenic bacteria: , , and All vaginal microbiota and efficiently colonized the 3-D surface, localizing to crevices of the EEC model and interacting with multiple adjacent cells simultaneously. However, only infection with pathogenic and not infection with the other bacteria tested significantly induced proinflammatory mediators and significant ultrastructural changes to the host cells. The latter observation is consistent with clinical findings and illustrated the functional specificity of our system. Additionally, we highlighted the utility of the 3-D EEC model for the study of the pathogenesis of using a well-characterized Δ mutant. Overall, this study demonstrates that the human 3-D EEC model is a robust tool for studying host-microbe interactions and bacterial pathogenesis in the upper FRT.
来自女性下生殖道（FRT）的致病细菌上行至子宫内膜并定殖，与许多妇产科健康并发症相关。为了研究这些宿主与微生物的相互作用，我们利用HEC-1A细胞系和旋转壁式生物反应器（RWV）技术开发了一种人三维（3-D）子宫内膜上皮细胞（EEC）模型。我们的模型由3-D EEC聚集体组成，概括了人子宫内膜上皮组织的几个功能/结构特征，包括细胞分化、连接复合体/桥粒和微绒毛的存在，以及膜相关粘蛋白和Toll样受体（TLR）的产生。通过将EEC聚集体暴露于病毒和细菌产物来评估TLR功能。用聚肌苷酸-聚胞苷酸（poly(I·C)）和鞭毛蛋白处理可显著以剂量依赖方式诱导促炎介质，但用合成脂蛋白（成纤维细胞刺激脂蛋白1 [FSL-1]）或脂多糖（LPS）处理则不然。为了模拟上行感染，我们用共生菌和致病菌感染EEC聚集体： 、 和 。所有阴道微生物群和 均有效地定殖于3-D表面，定位于EEC模型的缝隙处并同时与多个相邻细胞相互作用。然而，只有致病性 的感染而非其他测试细菌的感染显著诱导促炎介质并使宿主细胞发生显著超微结构变化。后一观察结果与临床发现一致，并说明了我们系统的功能特异性。此外，我们利用特征明确的Δ突变体强调了3-D EEC模型在研究 发病机制方面的实用性。总体而言，本研究表明人3-D EEC模型是研究上FRT中宿主与微生物相互作用及细菌发病机制的有力工具。
