Pseudomonas aeruginosa induction of apoptosis in respiratory epithelial cells -: Analysis of the effects of cystic fibrosis transmembrane conductance regulator dysfunction and bacterial virulence factors
Pseudomonas aeruginosa induction of apoptosis in respiratory epithelial cells -: Analysis of the effects of cystic fibrosis transmembrane conductance regulator dysfunction and bacterial virulence factors
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DOI：
10.1165/ajrcmb.23.3.4098
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发表时间：
2000-09-01
影响因子：
6.4
通讯作者：
Prince, A
中科院分区：
文献类型：
--
作者：
Rajan, S;Cacalano, G;Prince, A
气道上皮细胞可通过激活几种信号通路对感染作出反应。我们检测了铜绿假单胞菌PAO1在正常细胞和几种囊性纤维化（CF）和校正细胞系中的诱导凋亡作用。共聚焦显微镜显示，顶端ZO-1染色证实，连接紧密的单层上皮细胞完全抵抗pao1诱导的细胞凋亡。相反，不形成紧密连接的细胞系如9HTEo（-）细胞易受影响，暴露于PAO1 6小时后，50%的细胞凋亡，不同机制（运输突变、调节结构域过表达或反义结构）引起的CF跨膜电导调节（CFTR）功能障碍没有改变凋亡率。在末端脱氧核糖核苷转移酶介导的脱氧尿苷三磷酸-生物素镍末端标记检测PAO1感染cftr -/-或对照小鼠气道细胞凋亡方面也没有明显差异。即使在易感上皮细胞中，细菌表达特异性粘附素、完整的脂多糖和功能性III型分泌系统都是引起细胞凋亡所必需的。与其他粘膜表面不同，气道上皮对细胞凋亡具有高度的抗性，并且只有当适当的上皮条件存在以及具有完全毒力的铜绿假单胞菌能够协调表达粘附素和细胞毒素时，这种反应才会被激活。
Airway epithelial cells can respond to infection by activating several signaling pathways. We examined the induction of apoptosis in response to Pseudomonas aeruginosa PAO1 in normal cells and several cystic fibrosis (CF) and corrected cell lines. Epithelial cells in monolayers with tight junctions, confirmed by apical ZO-1 staining demonstrated by confocal microscopy, were entirely resistant to PAO1-induced apoptosis. In contrast, cell lines such as 9HTEo(-) cells that do not form tight junctions were susceptible, with 50% of the population apoptotic after 6 h of exposure to PAO1, CF transmembrane conductance regulator (CFTR) dysfunction caused by different mechanisms (trafficking mutations, overexpression of the regulatory domain or antisense constructs) did not alter rates of apoptosis, nor were differences apparent in terminal deoxyribonucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick-end labeling detection of apoptotic airway cells from PAO1 infected cftr -/- or control mice. Bacterial expression of specific adhesins, complete lipopolysaccharide, and a functional type III secretion system were all necessary to evoke apoptosis even in susceptible epithelial cells. Unlike other mucosal surfaces, the airway epithelium is highly resistant to apoptosis, and this response is activated only when the appropriate epithelial conditions are present as well as fully virulent P. aeruginosa capable of coordinately expressing both adhesins and cytotoxins.