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基础科学方向。犬冠状病毒在极化上皮细胞中的进入与释放。

Basic science track. Entry and release of canine coronavirus from polarized epithelial cells.

作者信息

Pratelli Annamaria

机构信息

Department of Public Health and Animal Sciences, Faculty of Veterinary Medicine, Valenzano, Bari, Italy.

出版信息

New Microbiol. 2011 Jan;34(1):25-32. Epub 2011 Jan 15.

Abstract

Virus entry into and release from epithelial cells are polarized as a result of the distribution of the viral receptors. In order to establish the polarity of entry and release of CCoV from epithelial cells, the interactions of the virus with A72 and CrFK cells grown on permeable supports was evaluated, and the amount of infective virus in the apical and in the basolateral media was determined and compared. Infection of A72 cells after different times post seeding demonstrated that CCoV grow after infection from both apical and basolateral sides. In CrFK cells, CCoV was observed in both compartments only in the later phase of the infection. To establish the reciprocal binding of CCoV on plasma membrane, A72 cells on a permeable support were preincubated with a mAb specific for CCoV. Infection from the apical side was blocked by mAb applied to that side; in contrast, such treatment on the basolateral side had no effect on the infectious process. Similarly, the low levels of CCoV observed after basolateral exposure to virus was abolished following mAb treatment of that side. The identification of CCoV into the basolateral medium could play an important role in the viral pathogenesis.

摘要

由于病毒受体的分布,病毒进入上皮细胞以及从上皮细胞释放的过程是极化的。为了确定犬冠状病毒(CCoV)从上皮细胞进入和释放的极性,评估了病毒与生长在可渗透支持物上的A72细胞和猫肾传代细胞(CrFK细胞)的相互作用,并测定和比较了顶端和基底外侧培养基中感染性病毒的量。接种后不同时间对A72细胞进行感染,结果表明CCoV感染后从顶端和基底外侧都能生长。在CrFK细胞中,仅在感染后期在两个区室中都观察到了CCoV。为了确定CCoV在质膜上的相互结合情况,将生长在可渗透支持物上的A72细胞与针对CCoV的单克隆抗体(mAb)进行预孵育。应用于顶端侧的mAb可阻断从该侧的感染;相反,在基底外侧进行这种处理对感染过程没有影响。同样,在基底外侧暴露于病毒后观察到的低水平CCoV在对该侧进行mAb处理后消失。CCoV进入基底外侧培养基可能在病毒发病机制中起重要作用。

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