Guibinga G H, Lochmuller H, Massie B, Nalbantoglu J, Karpati G, Petrof B J
Department of Medicine, Royal Victoria Hospital, and Meakins-Christie Laboratories, McGill University, Montreal, Quebec, Canada H3A 1A1.
J Virol. 1998 Jun;72(6):4601-9. doi: 10.1128/JVI.72.6.4601-4609.1998.
Recombinant adenovirus vectors (AdV) have been considered a potential vehicle for performing gene therapy in patients suffering from Duchenne muscular dystrophy but are limited by a cellular and humoral immune response that prevents long-term transgene expression as well as effective transduction after AdV readministration. Conventional immunosuppressive agents such as cyclosporine and FK506, which act by interfering with CD3-T-cell receptor-mediated signaling via calcineurin, are only partially effective in reversing these phenomena and may also produce substantial organ toxicity. We hypothesized that activation of redundant T-cell activation pathways could limit the effectiveness of these drugs at clinically tolerable doses. Therefore, we have tested the ability of immunomodulatory immunoglobulins (Ig) with different modes of action to facilitate AdV-mediated gene transfer to adult dystrophic (mdx) mice. When used in isolation, immunomodulatory Ig (anti-intercellular adhesion molecule-1, anti-leukocyte function-associated antigen-1, anti-CD2, and CTLA4Ig) were only mildly effective in mitigating cellular and/or humoral immunity against adenovirus capsid proteins and the therapeutic transgene product, dystrophin. However, the combination of FK506 plus CTLA4Ig abrogated the immune response against adenovirus proteins and dystrophin to a degree not achievable with the use of either agent alone. At 30 days after AdV injection, >90% of myofibers could be found to express dystrophin with little or no evidence of a cellular immune response against transduced fibers. In addition, the humoral immune response was markedly suppressed, and this was associated with increased transduction efficiency following vector readministration. These data suggest that by facilitating both primary and secondary transduction after AdV administration, combined targeting of CD3-T-cell receptor-mediated signaling via calcineurin and the B7:CD28 costimulatory pathway could greatly increase the potential utility of AdV-mediated gene transfer as a therapeutic modality for genetic diseases such as Duchenne muscular dystrophy that will require long-term transgene expression and repeated vector delivery.
重组腺病毒载体（AdV）被认为是对患有杜氏肌营养不良症的患者进行基因治疗的一种潜在载体，但受到细胞和体液免疫反应的限制，这种免疫反应会阻止长期转基因表达以及AdV再次给药后的有效转导。传统的免疫抑制剂，如环孢素和FK506，它们通过钙调神经磷酸酶干扰CD3 - T细胞受体介导的信号传导起作用，在逆转这些现象方面仅部分有效，并且还可能产生严重的器官毒性。我们推测冗余T细胞激活途径的激活可能会限制这些药物在临床可耐受剂量下的有效性。因此，我们测试了具有不同作用方式的免疫调节性免疫球蛋白（Ig）促进AdV介导的基因转移至成年营养不良（mdx）小鼠的能力。单独使用时，免疫调节性Ig（抗细胞间黏附分子 - 1、抗白细胞功能相关抗原 - 1、抗CD2和CTLA4Ig）在减轻针对腺病毒衣壳蛋白和治疗性转基因产物肌营养不良蛋白的细胞和/或体液免疫方面仅具有轻微效果。然而，FK506与CTLA4Ig的联合使用在一定程度上消除了针对腺病毒蛋白和肌营养不良蛋白的免疫反应，这是单独使用任何一种药物都无法实现的。在AdV注射后30天，>90%的肌纤维可被发现表达肌营养不良蛋白，几乎没有或没有针对转导纤维的细胞免疫反应的证据。此外，体液免疫反应明显受到抑制，这与载体再次给药后转导效率的提高有关。这些数据表明，通过促进AdV给药后的初次和二次转导，联合靶向通过钙调神经磷酸酶的CD3 - T细胞受体介导的信号传导和B7:CD28共刺激途径，可以大大增加AdV介导的基因转移作为杜氏肌营养不良症等需要长期转基因表达和重复载体递送的遗传疾病治疗方式的潜在效用。
