Long-term expression and repeated administration of AAV type 1, 2 and 5 vectors in skeletal muscle of immunocompetent adult mice
Long-term expression and repeated administration of AAV type 1, 2 and 5 vectors in skeletal muscle of immunocompetent adult mice
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DOI：
10.1038/sj.gt.3302766
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发表时间：
2006-09-01
期刊：
影响因子：
5.1
通讯作者：
Douar, A. M.
中科院分区：
文献类型：
--
作者：
Riviere, C.;Danos, O.;Douar, A. M.
在许多动物物种中，腺相关病毒（AAV）载体促进长期基因转移到肌肉中。增加的表达水平可以通过使用替代血清型与重复给药的组合获得。在这里，我们比较了基于血清型1、2和5的AAV载体在免疫功能正常的小鼠中的作用，并评估了相同（再给药）或不同（交叉给药）基于血清型载体多次给药的可行性。一项为期1年的剂量反应研究证实了重组(r) AAV1的优越性，在小鼠骨骼肌中的转导水平分别比rAAV2和rAAV5高5至10倍。反复给药表明，在第一次给药rAAV2或rAAV5后，第二次注射rAAV1可以增加基因转移水平。用载体编码不同基因的再给药研究允许仅从第二个载体评估基因表达。当动物先前暴露于相同血清型时，所有三种raav均被抑制。相反，在交叉给药中，第二载体的基因表达没有明显变化。在初始暴露后，所有三种血清型的病毒衣壳都引起了体液免疫应答。中和抗体（NAB）水平与载体注射剂量相关。在体外没有观察到一种血清型NAB对另一种血清型的显著交叉反应性。这些数据首次提供了rAAV1、rAAV2和rAAV5在肌肉中的再给药和交叉给药的直接比较评价，并进一步表明rAAV1在交叉给药时能够传导肌肉组织。
Adeno-associated viral (AAV) vectors promote long-term gene transfer into muscle in many animal species. Increased expression levels may be obtained by using alternative serotypes in combination with repeated administrations. Here we compared AAV vectors based on serotypes 1, 2 and 5 in immunocompetent mice and assessed the feasibility of multiple administrations of either identical (readministration) or different (cross-administration) serotype-based vectors. A 1-year-long dose-response study confirmed the superiority of recombinant (r) AAV1, achieving transduction levels 5 to 10-fold higher than rAAV2 and rAAV5 in mouse skeletal muscle, respectively. Repeated administration demonstrated that increased gene transfer level was achieved with a second injection of rAAV1 following the first administration of rAAV2 or rAAV5. A readministration study with a vector encoding a different gene allowed the evaluation of gene expression from the second vector only. All three rAAVs were inhibited when the animals were previously exposed to the same serotype. In contrast, no significant change in gene expression from the second vector was observed in cross-administration. A humoral immune response was elicited against the viral capsid for all three serotypes following the initial exposure. Neutralizing antibody ( NAB) levels correlated with the vector dose injected. No significant cross-reactivity of NAB from a given serotype toward another was observed in vitro. These data provide the first direct comparative evaluation of re- and cross-administration of rAAV1, rAAV2 and rAAV5 in muscle, and further indicate that rAAV1 is capable of transducing muscle tissue when cross-administered.