An in vitro tissue culture bilayer model to examine early events in Mycobacterium tuberculosis infection
An in vitro tissue culture bilayer model to examine early events in Mycobacterium tuberculosis infection
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DOI：
10.1128/iai.67.2.653-658.1999
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发表时间：
1999-02-01
影响因子：
3.1
通讯作者：
Quinn, FD
中科院分区：
文献类型：
--
作者：
Birkness, KA;Deslauriers, M;Quinn, FD
一个组织培养双层系统，模仿早期肺泡感染结核分枝杆菌的某些方面被开发。该模型结合了由微孔膜分隔的人肺上皮II型肺细胞（A549）（上腔）和内皮细胞（下腔）层。这种结构可以观察和量化细菌通过两层的通道，观察细菌与各种细胞类型的相互作用，并检查免疫细胞募集到感染部位的基本机制。在将10(7)个微生物加入上腔后，我们在显微镜下观察到附着在肺细胞上和肺细胞内的大量细菌，我们通过活细胞计数确定一小部分接种物（0.02至0.43%）通过双层进入下腔。当外周血单个核细胞加入下腔时，显微镜检查显示单个核细胞通过双层迁移到顶表面，在那里它们与肺细胞上的分枝杆菌有关。双层系统增加的复杂性为更精确地定义各种肺细胞类型在早期结核病发病机制中的作用提供了机会。
A tissue culture bilayer system that mimics some aspects of early alveolar infection by Mycobacterium tuberculosis was developed. This model incorporates human lung epithelial type II pneumocyte (A549) (upper chamber) and endothelial cell (lower chamber) layers separated by a microporous membrane. This construction makes it possible to observe and quantify the passage of bacteria through the two layers, to observe the interaction of the bacteria with the various cell types, and to examine the basic mechanisms of immune cell recruitment to the site of infection. After 10(7) organisms were added to the upper chamber we microscopically observed large numbers of bacteria attached to and within the pneumocytes and we determined by viable-cell counting that a small percentage of the inoculum (0.02 to 0.43%) passed through the bilayer into the lower chamber. When peripheral blood mononuclear cells were added to the lower chamber, microscopic examination indicated a migration of the mononuclear cells through the bilayer to the apical surface, where they were seen associated with the mycobacteria on the pneumocytes. The added complexity of the bilayer system offers an opportunity to define more precisely the roles of the various lung cell types in the pathogenesis of early tuberculosis.