Polyinosinic acid enhances delivery of adenovirus vectors in vivo by preventing sequestration in liver macrophages
Polyinosinic acid enhances delivery of adenovirus vectors in vivo by preventing sequestration in liver macrophages
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DOI：
10.1099/vir.0.83495-0
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发表时间：
2008-05-01
影响因子：
3.8
通讯作者：
Bellu, Anna Rita
中科院分区：
文献类型：
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作者：
Haisma, Hidde J.;Kamps, Jan A. A. M.;Bellu, Anna Rita
腺病毒因其优越的体内基因转移效率而成为基因治疗的首选载体之一。然而，在全身给药后，腺病毒被肝脏优先隔离，导致腺病毒介导的转基因在靶组织中的表达减少。在肝脏中，Kupffer细胞负责腺病毒的降解，并有助于炎症反应。由于存在于Kupffer细胞上的清道夫受体负责消除血源性病原体，我们研究了这些受体在清除腺病毒载体中的可能含义。多肌苷酸[poly(I)]是一种清道夫受体a配体，分析了其在巨噬细胞中特异性抑制腺病毒摄取的能力。在体外研究中，在病毒感染前添加poly(I)导致腺病毒诱导的基因表达在J774巨噬细胞系和原代Kupffer细胞中特异性抑制。在体内实验中，预先注射聚(I)可使血液中循环的腺病毒颗粒数量瞬间增加10倍。因此，与未接受poly(I)的动物相比，在不同组织中测量的转基因表达水平增强了（5- 15倍）。最后，通常作为全身性腺病毒给药的结果而发生的库普弗细胞坏死被聚(I)的使用所阻止。聚(I)处理后，肝酶水平无毒性。从我们的数据，我们得出结论，聚(I)可以阻止腺病毒被肝巨噬细胞隔离。这些结果表明，通过抑制腺病毒被库普弗细胞摄取，有可能减少病毒载体的剂量，以减少肝毒性作用，并提高靶组织中的转基因表达水平。在系统基因治疗应用中，这将对靶向腺病毒载体的发展产生重大影响。
Adenovirus is among the preferred vectors for gene therapy because of its superior in vivo gene-transfer efficiency. However, upon systemic administration, adenovirus is preferentially sequestered by the liver, resulting in reduced adenovirus-mediated transgene expression in targeted tissues. In the liver, Kupffer cells are responsible for adenovirus degradation and contribute to the inflammatory response. As scavenger receptors present on Kupffer cells are responsible for the elimination of blood-borne pathogens, we investigated the possible implication of these receptors in the clearance of the adenovirus vector. Polyinosinic acid [poly(I)], a scavenger receptor A ligand, was analysed for its capability to inhibit adenovirus uptake specifically in macrophages. In in vitro studies, the addition of poly(I) before virus infection resulted in a specific inhibition of adenovirus-induced gene expression in a J774 macrophage cell line and in primary Kupffer cells. In in vivo experiments, pre-administration of poly(I) caused a 10-fold transient increase in the number of adenovirus particles circulating in the blood. As a consequence, transgene expression levels measured in different tissues were enhanced (by 5- to 15-fold) compared with those in animals that did not receive poly(I). Finally, necrosis of Kupffer cells, which normally occurs as a consequence of systemic adenovirus administration, was prevented by the use of poly(I). No toxicity, as measured by liver-enzyme levels, was observed after poly(I) treatment. From our data, we conclude that poly(I) can prevent adenovirus sequestration by liver macrophages. These results imply that, by inhibiting adenovirus uptake by Kupffer cells, it is possible to reduce the dose of the viral vector to diminish the liver-toxicity effect and to improve the level of transgene expression in target tissues. In systemic gene-therapy applications, this will have great impact on the development of targeted adenoviral vectors.