Infection of A549 cells with a recombinant adenovirus vector induces ICAM-1 expression and increased CD-18-dependent adhesion of activated neutrophils.
Infection of A549 cells with a recombinant adenovirus vector induces ICAM-1 expression and increased CD-18-dependent adhesion of activated neutrophils.
复制标题
用重组腺病毒载体感染 A549 细胞可诱导 ICAM-1 表达并增加活化中性粒细胞的 CD-18 依赖性粘附。
DOI：
10.1089/hum.1996.7.14-1669
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发表时间：
1996
影响因子：
4.2
通讯作者：
Trapnell,BC
中科院分区：
文献类型：
--
作者：
Stark,JM;Amin,RS;Trapnell,BC
囊性纤维化（CF）和哮喘患者的大量肺部急性加重与呼吸道病毒感染有关。这一过程的分子介导者开始被理解。病毒感染呼吸道上皮细胞可导致细胞间粘附分子-1（ICAM-1）（炎性细胞粘附和活化的配体）和多种促炎细胞因子的产生。目前正在评估来源于人腺病毒（AV）的人基因治疗载体将CF跨膜调节因子（CFTR）基因递送至CF患者的气道上皮。然而，使用这些AV载体的动物模型研究表明AV暴露后肺部炎症。使用一个体外模型，我们研究了这一假设，即呼吸道上皮细胞暴露于AV载体导致ICAM-1基因表达上调。使用复制缺陷型第一代AV载体进行感染。将A549细胞（人肺腺癌细胞系）以50-150空斑形成单位/细胞的感染复数暴露于AV（导致在暴露后48小时>90%的细胞表达报告基因）。采用酶免疫分析、流式细胞术和北方印迹分析，在病毒暴露后的时间间隔测量ICAM-1表达。在载体暴露后96小时，细胞结合的ICAM-1显著增加，是对照的2至4倍，p< 0.001）。AV暴露的A549细胞还支持AV暴露后96小时活化的中性粒细胞的粘附水平增加（四倍于对照，p< 0.001），其被CD 18抗体阻断。A549单层的AV暴露增加生物活性ICAM-1的表达。减少宿主细胞对复制缺陷型AV载体的促炎反应的策略可能会提高其基因治疗的安全性。
A significant number of pulmonary exacerbations in patients with cystic fibrosis (CF) and asthma are associated with respiratory virus infections. The molecular mediators of this process are beginning to be understood. Viral infection of respiratory epithelial culturesin vitroleads to the production of intercellular adhesion molecule-1 (ICAM-1) (a ligand for inflammatory cell adhesion and activation) and a number of proinflammatory cytokines. Human gene therapy vectors derived from human adenoviruses (AV) are currently under evaluation for CF transmembrane regulator (CFTR) gene delivery to the airway epithelium of CF patients. However, studies in animal models using these AV vectors demonstrate pulmonary inflammation following AV exposure. Using anin vitromodel, we examined the hypothesis that exposure of respiratory epithelial cells to AV vectors results in upregulation of ICAM-1 gene expression. Infections were performed using a replication-deficient, first-generation AV vector. A549 cells (a human pulmonary adenocarcinoma cell line) were exposed to AV at multiplicity of infection of 50–150 plaque-forming units/cell (resulting in >90% of cells expressing the reporter gene by 48 hr following exposure). Measurements of ICAM-1 expression were made at time intervals following virus exposure using enzyme immunoassay, flow cytometry, and Northern blot analysis. Cell-bound ICAM-1 was significantly increased 96 hr following vector exposure, two to four times control,p< 0.001). The AV-exposed A549 cells also supported increased levels of adhesion of activated neutrophils 96 hr following AV exposure (four times control,p< 0.001) that was blocked by antibody to CD18. AV exposure of A549 monolayers increases expression of biologically active ICAM-1. Strategies to minimize host cellular proinflammatory responses to the replication-deficient AV vectors may improve their safety for gene therapy.