Polarity influences the efficiency of recombinant adenoassociated virus infection in differentiated airway epithelia
Polarity influences the efficiency of recombinant adenoassociated virus infection in differentiated airway epithelia
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DOI：
10.1089/hum.1998.9.18-2761
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发表时间：
1998-12-10
影响因子：
4.2
通讯作者：
Engelhardt, JF
中科院分区：
文献类型：
--
作者：
Duan, DS;Yue, YP;Engelhardt, JF
为了更好地理解限制rAAV在肺中转导的机制，我们评估了rAAV感染在极化的原代气道上皮培养物中的几个独特特征。发现rAAV覆盖气道上皮细胞的基底外侧表面比顶膜有效200倍。膜感染的这些差异与顶端硫酸乙酰肝素蛋白聚糖（AAV-2受体）和病毒结合的丰度相关。UV照射增强rAAV转导大于20倍，只有当病毒被施加到顶端膜。紫外线照射的原代培养物的超微结构分析表明，在暴露于25 J/m2的紫外线后，微绒毛结构发生了显着变化。虽然病毒结合和硫酸乙酰肝素蛋白聚糖的丰度没有增加在紫外线照射后的顶端膜，增加受体独立的荧光珠的内吞作用被认为是在紫外线照射后的顶端膜。我们推测，与顶端膜特异性途径的病毒进入，和/或处理病毒的细胞核，可能会改变紫外线照射后的内吞过程。有趣的是，紫外线照射对rAAV从基底外侧膜的转导具有抑制作用，这与基底膜处硫酸乙酰肝素蛋白聚糖丰度的降低相关。总之，这些研究结果表明，独立的病毒转导途径可能发生在极化气道上皮细胞的顶端和基底外侧室。
To better understand mechanisms that limit rAAV transduction in the lung, we have evaluated several unique features of rAAV infection in polarized primary airway epithelial cultures. rAAV was found to transduce the basolateral surface of airway epithelia 200-fold more efficiently than the apical membrane. These differences in membrane infection correlated with the abundance of apical heparan sulfate proteoglycan (AAV-2 receptor) and virus binding. UV irradiation augmented rAAV transduction greater than 20-fold, only when virus was applied to the apical membrane. Ultrastructural analysis of UV-irradiated primary cultures demonstrated significant changes in microvilli architecture following exposure to 25 J/m(2) UV. Although virus binding and the abundance of heparan sulfate proteoglycan were not increased at the apical membrane following UV irradiation, increased receptor-independent endocytosis of fluorescent beads was seen at the apical membrane following UV irradiation. We hypothesize that endocytotic processes associated with apical membrane-specific pathways of viral entry, and/or processing of virus to the nucleus, may be altered following UV irradiation. Interestingly, UV irradiation had an inhibitory effect on rAAV transduction from the basolateral membrane, which correlated with a decrease in the abundance of heparan sulfate proteoglycan at the basal membrane. In summary, these findings suggest that independent pathways of viral transduction may occur in the apical and basolateral compartments of polarized airway epithelia.