Effects of nitric oxide on Pseudomonas aeruginosa infection of epithelial cells from a human respiratory cell line derived from a patient with cystic fibrosis
Effects of nitric oxide on Pseudomonas aeruginosa infection of epithelial cells from a human respiratory cell line derived from a patient with cystic fibrosis
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DOI：
10.1128/iai.71.5.2341-2349.2003
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发表时间：
2003-05-01
影响因子：
3.1
通讯作者：
Evans, TJ
中科院分区：
文献类型：
--
作者：
Darling, KEA;Evans, TJ
囊性纤维化（CF）的特征是气道炎症和慢性细菌性肺部感染，最常见的是铜绿假单胞菌，一种机会性人类病原体。尽管在CF患者中观察到持续的气道炎症，尽管吞噬细胞诱导型一氧化氮合酶（iNOS）的产生上调，但呼吸道上皮中iNOS的表达显著降低。鉴于NO的抗菌作用，这可能有助于这种疾病的慢性气道感染。为了确定上皮来源的NO在气道防御铜绿假单胞菌中的作用，我们用该病原体的不同菌株以低感染复数感染来自CF患者的分化的人支气管上皮细胞（CFBE 410-细胞）。使用转染人iNOS cDNA的细胞，我们研究了NO对铜绿假单胞菌复制、粘附和内化的影响。与对照值相比，铜绿假单胞菌对iNOS表达细胞的粘附减少了44%至72%（P = 0.02）。铜绿假单胞菌对这些细胞的绝对摄取减少了44%，但以粘附细菌的百分比表示的摄取与对照摄取没有差异。在感染后晚期，表达iNOS的细胞内铜绿假单胞菌的存活率降低（P = 0.034）。NO的产生并不改变宿主611的生存能力。NO的产生减少了铜绿假单胞菌对人支气管上皮细胞的粘附，并增强了对内化细菌的杀伤，这表明CF患者中上皮iNOS的缺乏可能有助于铜绿假单胞菌感染和定植。
Cystic fibrosis (CF) is characterized by airway inflammation and chronic bacterial lung infection, most commonly with Pseudomonas aeruginosa, an opportunistic human pathogen. Despite the persistent airway inflammation observed in patients with CF, although phagocyte inducible nitric oxide synthase (iNOS) production is upregulated, expression of iNOS in the respiratory epithelium is markedly reduced. Given the antimicrobial action of NO, this may contribute to the chronic airway infection of this disease. To define the role of epithelium-derived NO in airway defense against P. aeruginosa, we infected differentiated human bronchial epithellial cells derived from a patient with CF (CFBE41o- cells) with different strains of this pathogen at low multiplicities of infection. Using cells transfected with human iNOS cDNA, we studied the effect of NO on P. aeruginosa replication, adherence, and internalization. P. aeruginosa adherence to iNOS-expressing cells was reduced by 44 to 72% (P = 0.02) compared with control values. Absolute P. aeruginosa uptake into these cells was reduced by 44%, but uptake expressed as a percentage of adherent bacteria did not differ from the control uptake. Survival of P. aeruginosa within iNOS-expressing cells was reduced at late times postinfection (P = 0.034). NO production did not alter host 611 viability. NO production reduced P. aeruginosa adherence to human bronchial epithellial cells and enhanced killing of internalized bacteria, suggesting that a lack of epithelial iNOS in patients with CF may contribute to P. aeruginosa infection and colonization.