Casale T B, Mower D A, Carolan E J
Nebraska Medical Research Institute, Papillion 68046-4796, USA.
Exp Lung Res. 1998 Nov-Dec;24(6):709-19. doi: 10.3109/01902149809099591.
To better understand the mechanisms by which neutrophils migrate into the airways, we constructed a novel in vitro model system with human umbilical vein endothelial cell (HUVE) monolayers grown on top of permeable filters and human lung Type II-like alveolar epithelial cell (A549) monolayers grown on the undersurface of the filters. The sequential migration of human neutrophils through the endothelium (apical to basal movement) and subsequently through the epithelium (basal to apical movement) in response to a stimulus located basally to the epithelium was measured. We found that the neutrophil chemoattractants, formylmethionylleucylphenylalanine (FMLP), leukotriene B4 (LTB4), and interleukin-8 (IL-8), induced dose-responsive migration through the double monolayer-filter complex. The pattern of migration was similar to that observed through either a naked filter or single monolayer-filter complex. Maximal chemotaxis through the double monolayer-filter complex was observed by 3 hours. Thus, we have established an in vitro model system to examine the sequential migration of neutrophils through endothelium and the respiratory epithelium in a manner analogous to that occurring with an in vivo airway stimulus causing neutrophil-rich airway inflammatory responses.
为了更好地理解中性粒细胞迁移至气道的机制，我们构建了一种新型体外模型系统，该系统在可渗透滤器顶部生长人脐静脉内皮细胞（HUVE）单层，在滤器下表面生长人肺II型样肺泡上皮细胞（A549）单层。测量了人中性粒细胞响应位于上皮细胞基底侧的刺激，依次穿过内皮细胞（从顶端到基底移动），随后穿过上皮细胞（从基底到顶端移动）的过程。我们发现，中性粒细胞趋化因子，甲酰甲硫氨酰亮氨酰苯丙氨酸（FMLP）、白三烯B4（LTB4）和白细胞介素-8（IL-8），可诱导通过双层单层滤器复合物的剂量反应性迁移。迁移模式与通过裸滤器或单层滤器复合物观察到的相似。在3小时时观察到通过双层单层滤器复合物的最大趋化作用。因此，我们建立了一种体外模型系统，以类似于体内气道刺激引起富含中性粒细胞的气道炎症反应的方式，研究中性粒细胞依次通过内皮细胞和呼吸道上皮细胞的迁移过程。
