AAV transcytosis through barrier epithelia and endothelium
AAV transcytosis through barrier epithelia and endothelium
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DOI：
10.1016/j.ymthe.2005.11.007
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发表时间：
2006-03-01
影响因子：
12.4
通讯作者：
Chiorini, JA
中科院分区：
文献类型：
--
作者：
Di Pasquale, G;Chiorini, JA
有效地转导屏障上皮（如肺）是几种基因治疗应用的目标。然而，AAV-2的实验表明，这种类型的屏障上皮的转导是有限的。相反，其他血清型AAV转导屏障上皮并在整个组织中广泛播散。胞吞作用是蛋白质和病原体克服屏障层到达相反的细胞表面的过程。为了更好地了解AAV颗粒的进入途径及其穿透屏障上皮的能力，我们验证了一些屏障上皮在体外的有限转导或一些AAV血清型在体内通过组织的传播是由于胞吞作用的假设。我们的实验表明，依赖病毒可以通过胞吞作用穿透屏障细胞。该过程快速且血清型和细胞型特异性，可通过中和抗体、温度或胞吞作用的化学抑制剂阻断。在根尖向基底侧转运后分离的颗粒仍然被包裹，它们可以在体外穿透允许细胞系。此外，AAV-5用于胞吞作用的进入途径似乎独立于用于转导的途径。重要的是，病毒胞吞作用的抑制导致细胞内载体和转导的急剧增加。
To transduce efficiently barrier epithelia such as the lung is the goal of several gene therapy applications. However, experiments with AAV-2 suggest that transduction is limited in this type of barrier epithelia. In contrast, other serotypes of AAV transduce barrier epithelia and exhibit broad dissemination throughout the tissue. Transcytosis is a process by which proteins and pathogens overcome barrier layers to reach the opposite cell surface. To understand better the entry pathway of AAV particles and their ability to penetrate barrier epithelia, we tested the hypothesis that the limited transduction of some barrier epithelia in vitro or the spread of some AAV serotypes through tissue in vivo is due to transcytosis. Our experiments demonstrate that dependoviruses can penetrate barrier cells by transcytosis. The process is rapid as well as serotype and cell-type specific and can be blocked by neutralizing antibodies, temperature, or chemical inhibitors of transcytosis. The particles isolated following apical-to-basolateral transport are still encapsulated and they can transcluce permissive cell lines in vitro. Furthermore, the entry pathway used by AAV-5 for transcytosis appears to be independent of the one used for transduction. Importantly, inhibition of virus transcytosis results in a dramatic increase in intracellular vector and transduction.