Immune Responses to AAV in Canine Muscle Monitored by Cellular Assays and Noninvasive Imaging
Immune Responses to AAV in Canine Muscle Monitored by Cellular Assays and Noninvasive Imaging
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DOI：
10.1038/mt.2009.294
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发表时间：
2010-03-01
影响因子：
12.4
通讯作者：
Tapscott, Stephen J.
中科院分区：
文献类型：
--
作者：
Wang, Zejing;Storb, Rainer;Tapscott, Stephen J.
我们以前在血友病B的人体试验中证明，直接肌肉注射rAAV2或rAAV6可以产生强大的T细胞对病毒衣壳蛋白的反应，其他人已经证明对腺相关病毒(AAV)衣壳蛋白的细胞免疫与肝脏毒性和消除转基因表达是一致的。在这里，我们展示了特定AAV血清型的肝素结合能力不决定在狗肌肉注射后诱导T细胞反应的能力，并确定了AAV衣壳蛋白中由AAV注射激发的T细胞识别的多个表位。我们还证明，无创磁共振成像(MRI)可以准确地检测到犬肌肉注射rAAV后的局部炎症反应。这些研究表明，假型rAAV载体去除肝素结合活性将不足以消除免疫原性，并验证了在狗的临床前研究中，酶联免疫吸附斑点(ELISpot)分析和MRI在监测rAAV载体肌肉注射后的免疫和炎症反应方面的有效性。这些检测应该被纳入未来AAV基因治疗的人类临床试验中，以监测免疫反应。
We previously demonstrated that direct intramuscular injection of rAAV2 or rAAV6 in wild-type dogs resulted in robust T-cell responses to viral capsid proteins, and others have shown that cellular immunity to adeno-associated virus (AAV) capsid proteins coincided with liver toxicity and elimination of transgene expression in a human trial of hemophilia B. Here, we show that the heparin-binding ability of a given AAV serotype does not determine the induction of T-cell responses following intramuscular injection in dogs, and identify multiple epitopes in the AAV capsid protein that are recognized by T cells elicited by AAV injection. We also demonstrate that noninvasive magnetic resonance imaging (MRI) can accurately detect local inflammatory responses following intramuscular rAAV injection in dogs. These studies suggest that pseudotyping rAAV vectors to remove heparin-binding activity will not be sufficient to abrogate immunogenicity, and validate the utility of enzyme-linked immunosorbent spot (ELISpot) assay and MRI for monitoring immune and inflammatory responses following intramuscular injection of rAAV vectors in preclinical studies in dogs. These assays should be incorporated into future human clinical trials of AAV gene therapy to monitor immune responses.