Chlamydia pneumoniae Infection in Polarized Epithelial Cell Lines
Chlamydia pneumoniae Infection in Polarized Epithelial Cell Lines
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DOI：
10.1128/iai.01456-09
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发表时间：
2010-03
影响因子：
3.1
通讯作者：
Liisa Törmäkangas;E. Markkula;K. Lounatmaa;M. Puolakkainen
中科院分区：
文献类型：
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作者：
Liisa Törmäkangas;E. Markkula;K. Lounatmaa;M. Puolakkainen
摘要我们建立了一种极化细胞培养模型，用于研究呼吸道常见病原体肺炎衣原体的致病性。ZO-1（一种紧密连接蛋白）和Na+K+ ATP酶（一种位于极化上皮细胞基底外侧膜的蛋白泵）的免疫荧光染色以及TER测量表明，过滤生长的Calu-3细胞，而不是A549细胞，在胶原蛋白包被的膜上生长时被极化。平板培养物和过滤培养物都感染了C。肺炎。在极化的Calu-3培养物中感染产生更多的C. pneumoniae基因组当量比感染在平坦的培养物。然而，这种后代的感染性不如在平板培养物中的后代。C.在过滤器生长的A549细胞中，在感染后6天检测到肺炎，表明发育周期比在平坦的A549培养物中观察到的发育周期慢。研究了放线菌酮对C. pneumoniae在极化细胞中的作用可以忽略不计。此外，极化的Calu-3细胞中的感染对强力霉素具有抗性，并且响应于C.肺炎感染。这些发现表明，衣原体的生长被改变的过滤生长的上皮细胞培养系统。C. pneumoniae感染的Calu-3细胞对强力霉素的抗性和细胞因子的极化分泌表明，该模型可用于检测上皮细胞对C.肺炎的感染，并且它可能更好地类似于呼吸道上皮细胞中的体内感染。
ABSTRACT We set up a polarized cell culture model to study the pathogenicity of a common respiratory tract pathogen, Chlamydia pneumoniae. Immunofluorescence staining of ZO-1 (a tight junction protein) and Na+K+ ATPase (a protein pump localized at the basolateral membrane in the polarized epithelial cells), as well as TER measurements, suggested that the filter-grown Calu-3 cells, but not the A549 cells, were polarized when grown on collagen-coated membranes. Both the flat and the filter-grown cultures were infected with C. pneumoniae. Infection in the polarized Calu-3 cultures produced more C. pneumoniae genome equivalents than infection in the flat cultures. However, this progeny was not as infective as that in the flat cultures. The maximum amount of C. pneumoniae was detected at 6 days postinfection in the filter-grown A549 cells, indicating a slower developmental cycle than that observed in the flat A549 cultures. The effect of cycloheximide on the growth of C. pneumoniae in the polarized cells was negligible. Furthermore, the infection in the polarized Calu-3 cells was resistant to doxycycline, and several cytokines were released mainly on the apical side of the polarized cells in response to C. pneumoniae infection. These findings indicate that the growth of chlamydiae was altered in the filter-grown epithelial culture system. The diminished production of infective progeny of C. pneumoniae, together with the resistance to doxycycline and polarized secretion of cytokines from the infected Calu-3 cells, suggests that this model is useful for examining epithelial cell responses to C. pneumoniae infection, and it might better resemble in vivo infection in respiratory epithelial cells.