Lopez-Souza N, Dolganov G, Dubin R, Sachs L A, Sassina L, Sporer H, Yagi S, Schnurr D, Boushey H A, Widdicombe J H
Department of Human Physiology, University of California-Davis, Davis CA 95616-8664, USA.
Am J Physiol Lung Cell Mol Physiol. 2004 Feb;286(2):L373-81. doi: 10.1152/ajplung.00300.2003.
Virtually all in vitro studies of the effects of rhinovirus on human airway epithelium have used cells grown under conditions known to produce low levels of differentiation. The relevance of the results to native epithelium is questionable. Here we grew primary cultures of human tracheal or nasal epithelium under three conditions. One condition produced pseudostratified, mucociliary cells virtually indistinguishable from native epithelium. The other two conditions produced undifferentiated squamous cells lacking cilia. Cells were infected for 6 h with rhinovirus-16. After a 24-h incubation period, we determined levels of viral RNA in the cells, numbers of infectious viral particles released in the mucosal medium, expression of a variety of epithelial cytokines and other proteins, release of IL-6 and IL-8, and transepithelial electrical resistance and voltage. After infection, levels of viral RNA in the poorly differentiated cells were 30 or 130 times those in the differentiated. Furthermore, expression of mRNA for inflammatory cytokines, release of infectious particles, and release of IL-6 and IL-8 were closely correlated with the degree of viral infection. Thus well-differentiated cells are much more resistant to viral infection and its functional consequences than are poorly differentiated cells from the same source.
几乎所有关于鼻病毒对人气道上皮细胞影响的体外研究都使用了在已知会产生低分化水平的条件下培养的细胞。这些结果与天然上皮细胞的相关性值得怀疑。在这里，我们在三种条件下培养了人气管或鼻上皮的原代培养物。一种条件产生了几乎与天然上皮细胞无法区分的假复层、具有黏液纤毛的细胞。另外两种条件产生了缺乏纤毛的未分化鳞状细胞。用鼻病毒-16感染细胞6小时。在24小时的孵育期后，我们测定了细胞中病毒RNA的水平、在黏膜培养基中释放的感染性病毒颗粒的数量、多种上皮细胞因子和其他蛋白质的表达、IL-6和IL-8的释放以及跨上皮电阻和电压。感染后，低分化细胞中病毒RNA的水平是分化细胞中的30倍或130倍。此外，炎性细胞因子mRNA的表达、感染性颗粒的释放以及IL-6和IL-8的释放与病毒感染程度密切相关。因此，与来自同一来源的低分化细胞相比，分化良好的细胞对病毒感染及其功能后果的抵抗力要强得多。
