Respiratory syncytial virus infection of human airway epithelial cells is polarized, specific to ciliated cells, and without obvious cytopathology
Respiratory syncytial virus infection of human airway epithelial cells is polarized, specific to ciliated cells, and without obvious cytopathology
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DOI：
10.1128/jvi.76.11.5654-5666.2002
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发表时间：
2002-06-01
影响因子：
5.4
通讯作者：
Pickles, RJ
中科院分区：
文献类型：
--
作者：
Zhang, LQ;Peeples, ME;Pickles, RJ
囊性纤维化（CF）肺疾病的基因治疗需要在腔内递送后将基因有效地转移到气道上皮细胞。到目前为止，大多数基因转移载体都没有提供所需的效率。尽管已知人呼吸道合胞病毒（RSV）（一种常见的呼吸道病毒）感染呼吸道上皮，但尚未确定感染机制和RSV靶向的上皮细胞类型。我们已经利用人原代气道上皮细胞培养，产生分化良好的假复层粘膜纤毛上皮细胞，以研究RSV是否通过内腔（顶端）表面感染气道上皮。当将表达绿色荧光蛋白（rgRSV）的重组RSV应用于顶端表面时，以高基因转移效率感染上皮细胞培养物，但在基底外侧接种后则不然。对RSV感染的细胞类型的分析表明，管腔柱状细胞，特别是纤毛上皮细胞，是RSV的靶向细胞，并且培养物在分化成纤毛表型时变得易受感染。除了通过顶端膜感染纤毛细胞外，RSV仅从顶端表面脱落，并通过纤毛搏动的运动传播到邻近的纤毛细胞。感染RSV的培养物的大体组织学检查显示没有明显的细胞病理学证据，表明在没有免疫应答的情况下RSV感染可以耐受>3个月。因此，rgRSV通过管腔表面有效转导气道上皮，并特异性靶向纤毛气道上皮细胞。由于rgRSV似乎突破了气道中其他基因转移载体遇到的内腔屏障，因此该病毒可能是开发用于CF肺病的基因转移载体的良好候选者。
Gene therapy for cystic fibrosis (CF) lung disease requires efficient gene transfer to airway epithelial cells after intralumenal delivery. Most gene transfer vectors so far tested have not provided the efficiency required. Although human respiratory syncytial virus (RSV), a common respiratory virus, is known to infect the respiratory epithelium, the mechanism of infection and the epithelial cell type targeted by RSV have not been determined. We have utilized human primary airway epithelial cell cultures that generate a well-differentiated pseudostratified mucociliary epithelium to investigate whether RSV infects airway epithelium via the lumenal (apical) surface. A recombinant RSV expressing green fluorescent protein (rgRSV) infected epithelial cell cultures with high gene transfer efficiency when applied to the apical surface but not after basolateral inoculation. Analyses of the cell types infected by RSV revealed that lumenal columnar cells, specifically ciliated epithelial cells, were targeted by RSV and that cultures became susceptible to infection as they differentiated into a ciliated phenotype. In addition to infection of ciliated cells via the apical membrane, RSV was shed exclusively from the apical surface and spread to neighboring ciliated cells by the motion of the cilial beat. Gross histological examination of cultures infected with RSV revealed no evidence of obvious cytopathology, suggesting that RSV infection in the absence of an immune response can be tolerated for >3 months. Therefore, rgRSV efficiently transduced the airway epithelium via the lumenal surface and specifically targeted ciliated airway epithelial cells. Since rgRSV appears to breach the lumenal barriers encountered by other gene transfer vectors in the airway, this virus may be a good candidate for the development of a gene transfer vector for CF lung disease.