Repeat transduction in the mouse lung by using adeno-associated virus vectors with different serotypes
Repeat transduction in the mouse lung by using adeno-associated virus vectors with different serotypes
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DOI：
10.1128/jvi.74.3.1524-1532.2000
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发表时间：
2000-02-01
影响因子：
5.4
通讯作者：
Miller, AD
中科院分区：
文献类型：
--
作者：
Halbert, CL;Rutledge, EA;Miller, AD
来源于腺相关病毒2型（AAV 2：2）的载体促进基因转移和长期表达;然而，我们发现，虽然基因表达在小鼠中可以持续至少8个月，但在兔中的表达在2个月内急剧减少。AAV 2介导的基因表达在人肺中的效率和持久性尚未确定，但似乎在个体的一生中重新施用是必要的。不幸的是，我们发现通过第二次施用AAV 2载体的转导被阻断，推测是由于响应于初次载体暴露而产生的中和抗体。在此，我们探索了用来自AAV血清型2、3和6的衣壳蛋白假型化的AAV 2载体用于在小鼠肺中再施用的用途。我们发现，AAV 6载体以至少与AAV 2假型载体一样高的速率转导肺部气道上皮和肺泡细胞，而AAV 3介导的转导率要低得多，AAV 6假型载体转导不受先前施用AAV 2或AAV 3载体的影响，并且AAV 2假型载体的转导不受先前施用AAV 6载体的影响，表明在小鼠中不产生针对AAV 2和AAV 6的交叉反应性中和抗体。有趣的是，虽然先前施用AAV 2载体完全阻断了第二AAV 2假型载体的转导，但先前施用AAV 6载体仅部分抑制了第二次施用AAV 6假型载体的转导。对从小鼠和人获得的血清的分析表明，AAV 6的免疫原性低于AAV 2，这有助于解释这一发现。这些结果支持单独和与AAV 2载体组合的用于肺基因治疗的AAV 6载体的开发。
Vectors derived from adeno-associated virus type 2 (AAV2:2) promote gene transfer and expression in the long; however, we have found that while gene expression can persist for at least 8 months in mice, it was reduced dramatically in rabbits over a period of 2 months. The efficiency and persistence of AAV2-mediated gene expression in the human lung have yet to be determined, but it seems likely that readministration will be necessary over the lifetime of an individual. Unfortunately, we have found that transduction by a second administration of an AAV2 vector is blocked, presumably due to neutralizing antibodies generated in response to the primary vector exposure. Here, we have explored the use of AAV2 vectors pseudotyped with capsid proteins from AAV serotypes 2, 3, and 6 for readministration in the mouse lung. We found that an AAV6 vector transduced airway epithelial and alveolar cells in the lung at rates that were at least as high as those of AAV2 pseudotype vectors, while transduction rates mediated by AAV3 were much lower, AAV6 pseudotype vector transduction was unaffected by prior administration of an AAV2 or AAV3 vector, and transduction by an AAV2 pseudotype vector was unaffected by prior AAV6 vector administration, showing that cross-reactive neutralizing antibodies against AAV2 and AAV6 are not generated in mice. Interestingly, while prior administration of an AAV2 vector completely blocked transduction by a second AAV2 pseudotype vector, prior administration of an AAV6 vector only partially inhibited transduction by a second administration of an AAV6 pseudotype vector. Analysis of sera obtained from mice and humans showed that AAV6 is less immunogenic than AAV2, which helps explain this finding. These results support the development of AAV6 vectors for lung gene therapy both alone and in combination with AAV2 vectors.