Endocytosis of adeno-associated virus type 5 leads to accumulation of virus particles in the Golgi compartment
Endocytosis of adeno-associated virus type 5 leads to accumulation of virus particles in the Golgi compartment
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DOI：
10.1128/jvi.76.5.2340-2349.2002
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发表时间：
2002-03-01
影响因子：
5.4
通讯作者：
Kartenbeck, J
中科院分区：
文献类型：
--
作者：
Bantel-Schaal, U;Hub, B;Kartenbeck, J
在讨论作为基因治疗载体的腺相关病毒（AAV）血清型中，AAV5型（AAV5）具有独特的优势。为了进一步了解aav5的特异性，我们利用免疫荧光、电镜和Western blot分析方法研究了在没有辅助病毒的情况下，野生型病毒在HeLa细胞中的进入途径。我们发现病毒在顶端细胞表面结合，特别是在微绒毛上，并且随着孵育时间的增加，病毒在细胞-细胞边界上积累。不同的结合动力学表明不同的结合性质在顶端和外侧的质膜。病毒的内吞作用主要由两个膜域的网格蛋白包被的囊泡完成；然而，在未涂覆的坑中也检测到颗粒。AAV5颗粒主要到达高尔基区，在反式高尔基网的池内、池内相关的囊泡内以及高尔基体的双染色体堆内均可检测到AAV5颗粒。这些数据表明，AAV5利用迄今尚未被描述为病毒进入途径的内吞途径。
Among the adeno-associated virus (AAV) serotypes which are discussed as vectors for gene therapy AAV type 5 (AAV5) represents a candidate with unique advantages. To further our knowledge on AAV5-specific characteristics, we studied the entry pathway of wild-type virus in HeLa cells in the absence of helper virus by immunofluorescence and electron microscopy and by Western blot analysis. We found virus binding at the apical cell surface, especially at microvilli and, with increasing incubation time, virus accumulation at cell-cell boundaries. The different binding kinetics suggest different binding properties at apical versus lateral plasma membranes. Endocytosis of viruses was predominantly by clathrin-coated vesicles from both membrane domains; however, particles were also detected in noncoated pits. AAV5 particles were mainly routed to the Golgi area, where they could be detected within cisternae of the trans-Golgi network and within vesicles associated with cisternae and with the dictyosomal stacks of the Golgi apparatus. These data suggest that AAV5 makes use of endocytic routes that have hitherto not been described as pathways for virus entry.