Immunity to adeno-associated virus-mediated gene transfer in a random-bred canine model of Duchenne muscular dystrophy
Immunity to adeno-associated virus-mediated gene transfer in a random-bred canine model of Duchenne muscular dystrophy
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DOI：
10.1089/hum.2006.093
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发表时间：
2007-01-01
影响因子：
4.2
通讯作者：
Kuhr, Christian S.
中科院分区：
文献类型：
--
作者：
Wang, Zejing;Allen, James M.;Kuhr, Christian S.
重组腺相关病毒（rAAV）介导的基因转移已经显示出用于治疗各种动物模型中的疾病的前景，包括杜氏肌营养不良症（DMD）的mdx小鼠模型。然而，在许多情况下，近交系小鼠的临床前研究未能成功预测人类临床反应。为了评估通过AAV介导的基因递送治疗人DMD患者的潜在临床效用，我们在随机繁殖的野生型狗中进行了一系列直接肌内注射。携带不同启动子-转基因盒的AAV血清型2和6如先前针对鼠研究所述产生并肌内施用。在不同时间点对注射部位进行活检，并分析转基因表达和免疫组织化学分析。与这些载体在鼠研究中的一般非免疫原性性质相反，AAV 2和AAV 6载体都引发了强有力的细胞免疫应答，而不管所表达的转基因、启动子的细胞特异性和注射的肌肉类型。通过各种方法进行的病毒纯化没有减少T细胞介导的浸润。我们的数据表明。AAV 2和AAV 6衣壳蛋白在注射到随机繁殖的狗的骨骼肌中时可以引发初级细胞免疫应答，并且表明在人类中对AAV载体的细胞免疫的可能性。
Recombinant adeno-associated virus (rAAV)-mediated gene transfer has shown promise for treating diseases in various animal models including the mdx mouse model of Duchenne muscular dystrophy (DMD). In many cases, however, preclinical studies in inbred mice have not successfully predicted human clinical responses. To assess the potential clinical utility of treating human DMD patients by AAV-mediated gene delivery, we performed a series of direct intramuscular injections in random-bred wild-type dogs. AAV serotypes 2 and 6 carrying different promoter-transgene cassettes were produced as previously described for murine studies and administered intramuscularly. The injection sites were biopsied at various time points and analyzed for transgene expression and immunohistochemical analysis. In contrast to the generally nonimmunogenic nature of these vectors in murine studies, both AAV2 and AAV6 vectors elicited robust cellular immune responses regardless of the transgene expressed, the cellular specificity of the promoter, and the muscle type injected. Viral purification by various methods did not diminish T cell-mediated infiltration. Our data indicate that. AAV2 and AAV6 capsid proteins can elicit primary cellular immune responses when injected into the skeletal muscle of random-bred dogs, and suggest the possibility of cellular immunity to AAV vectors in humans.