Efficient intrathymic gene transfer following in situ administration of a rAAV serotype 8 vector in mice and nonhuman primates.
Efficient intrathymic gene transfer following in situ administration of a rAAV serotype 8 vector in mice and nonhuman primates.
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在小鼠和非人灵长类动物中原位施用 rAAV 血清型 8 载体后进行有效的胸腺内基因转移。
DOI：
10.1038/mt.2008.272
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发表时间：
2009
期刊：
影响因子：
--
通讯作者：
Zimmermann,ValérieS
中科院分区：
文献类型：
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作者：
Moreau,Aurélie;Vicente,Rita;Dubreil,Laurence;Adjali,Oumeya;Podevin,Guillaume;Jacquet,Chantal;Deschamps,JackYves;Klatzmann,David;Cherel,Yan;Taylor,Naomi;Moullier,Philippe;Zimmermann,ValérieS
胸腺是T细胞发育的主要部位，在自我耐受的诱导中起着关键作用。我们之前已经证明，在小鼠胸腺内注射转基因表达的慢病毒载体(LV)可以纠正T细胞特异性的遗传缺陷。然而，胸腺细胞的转导效率不超过0.1-0.3%，我们在猕猴体内检测不到任何胸腺转导。因此，我们开始评估重组腺相关病毒(RAAV)载体转导小鼠和灵长类胸腺细胞的能力。在AAV血清2型衍生的单链AAV(SsAAV)和假型为1、2、4、5和8型衣壳蛋白的自身互补AAV(ScAAV)载体的活体管理中，scAAV2/8显著增强了小鼠胸腺的转导。此外，IT通过内窥镜引导将scAAV2/8颗粒注射到猕猴体内，导致了显著的基因转移。因此，在健康的动物中，胸腺基因转移不提供选择性优势，scAAV2/8是一种独特的工具，促进胸腺细胞的原位转移，随后将基因修饰的淋巴细胞输出到外周。
The thymus is the primary site of T-cell development and plays a key role in the induction of self-tolerance. We previously showed that the intrathymic (IT) injection of a transgene-expressing lentiviral vector (LV) in mice can result in the correction of a T cell–specific genetic defect. Nevertheless, the efficiency of thymocyte transduction did not exceed 0.1–0.3% and we were unable to detect any thymus transduction in macaques. As such, we initiated studies to assess the capacity of recombinant adeno-associated virus (rAAV) vectors to transduce murine and primate thymic cells.In vivoadministration of AAV serotype 2–derived single-stranded AAV (ssAAV) and self-complementary AAV (scAAV) vectors pseudotyped with capsid proteins of serotypes 1, 2, 4, 5, and 8 demonstrated that murine thymus transduction was significantly enhanced by scAAV2/8. Transgene expression was detected in 5% of thymocytes and, notably, transduced cells represented 1% of peripheral T lymphocytes. Moreover, IT administration of scAAV2/8 particles in macaques, by endoscopic-mediated guidance, resulted in significant gene transfer. Thus, in healthy animals, where thymic gene transfer does not provide a selective advantage, scAAV2/8 is a unique tool promoting thein situtransduction of thymocytes with the subsequent export of gene-modified lymphocytes to the periphery.