Humoral immunity to adeno-associated virus type 2 vectors following administration to murine and nonhuman primate muscle
Humoral immunity to adeno-associated virus type 2 vectors following administration to murine and nonhuman primate muscle
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DOI：
10.1128/jvi.74.5.2420-2425.2000
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发表时间：
2000-03-01
影响因子：
5.4
通讯作者：
Wilson, JM
中科院分区：
文献类型：
--
作者：
Chirmule, N;Xiao, WD;Wilson, JM
腺相关病毒(AAV)是一种能够长期表达基因的载体，在慢性病的治疗中是有用的，但在大多数治疗应用中，需要对载体进行重新管理。本研究描述了肌肉注射AAV衣壳蛋白后的体液免疫应答及其对载体管理的影响。对小鼠和恒河猴的研究表明，针对AAV衣壳蛋白的中和抗体的形成持续了一年多，然后逐渐减少，但这并没有阻止载体重新接种的效果。更详细的研究强烈表明，B细胞的反应依赖于T细胞。在一只生物相容的小鼠身上，用人类CD4的封闭抗体进一步评估了这一点，该抗体用于临床试验，其中内源性小鼠的CD4基因在功能上被人类的对应基因取代。用CD4抗体对CD4T细胞进行短暂的药物抑制可在CD4抗体的作用减弱后很长一段时间内防止抗载体反应；在不降低基因表达的情况下重新给药载体是可能的。我们的研究表明，尽管有必要暂时抑制CD4T细胞的功能以避免记忆B细胞的激活，但AAV可以在骨骼肌中实现真正持久的转基因表达(即延长基因植入和载体重新管理)。
Adeno-associated virus (AAV) is being developed as a vector capable of conferring long-term gene expression, which is useful in the treatment of chronic diseases, In most therapeutic applications, it is necessary to readminister the vector. This study characterizes the humoral immune response to AAV capsid proteins following intramuscular injection and its impact on vector readministration. Studies of mice and rhesus monkeys demonstrated the formation of neutralizing antibodies to AAV capsid proteins that persisted for over 1 year and then diminished, but this did not prevent the efficacy of vector readministration. More-detailed studies strongly suggested that the B-cell response was T cell dependent. This was further evaluated with a blocking antibody to human CD4, primatized for clinical trials, in a biologically compatible mouse in which the endogenous murine CD4 gene was functionally replaced with the human counterpart. Transient pharmacologic inhibition of CD4 T cells with CD4 antibody prevented an antivector response long after the effects of the CD4 antibody diminished; readministration of vector without diminution of gene expression was possible. Our studies suggest that truly durable transgene expression (i.e., prolonged genetic engraftment together with vector readministration) is possible with AAV in skeletal muscle, although it will be necessary to transiently inhibit CD4 T-cell function to avoid the activation of memory B cells.