Myoblast gene therapy in canine mucopolysaccharidosis. I: Abrogation by an immune response to alpha-L-iduronidase.
Myoblast gene therapy in canine mucopolysaccharidosis. I: Abrogation by an immune response to alpha-L-iduronidase.
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犬粘多糖贮积症的成肌细胞基因治疗。
DOI：
10.1089/hum.1996.7.13-1595
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发表时间：
1996
期刊：
影响因子：
--
通讯作者：
Kohn,DB
中科院分区：
文献类型：
--
作者：
Shull,RM;Lu,X;McEntee,MF;Bright,RM;Pepper,KA;Kohn,DB
对3只溶酶体酶α-L-艾杜糖醛酸酶缺乏的犬进行了针对肌肉的基因替代治疗。直接肌内注射编码α-L-艾杜糖醛酸酶基因cDNA的质粒未导致可检测的酶产生，但可能导致对艾杜糖醛酸酶蛋白的免疫致敏作用，而犬完全缺乏这种致敏作用。从骨骼肌活检组织中培养成肌细胞，并用含有肌肌酸激酶增强子控制下的犬基因的逆转录病毒载体转导。在培养的细胞中发生了数百倍的过度表达酶的生产，然而，在将培养的细胞重新引入犬中后，酶的生产迅速下降。在酶水平下降的同时，产生了针对艾杜糖醛酸酶的特异性免疫球蛋白G（IgG）抗体，这进一步与成肌细胞注射部位的细胞浸润相关。大多数炎性细胞是淋巴细胞和浆细胞，表明局部体液和细胞免疫反应的酶产生的肌肉细胞。最终给药后2-22周采集的组织的PCR分析显示，Neo和犬α-l-艾杜糖醛酸酶序列在成肌细胞百分比中持续存在，并逐渐降低。本研究结果在犬粘多糖样沉积症I模型中强调了这样一个事实，即对产生所需的、正常的但外源的蛋白质的细胞的免疫反应可能与对用于引入外源基因的病毒载体的反应一样阻碍基因治疗。
Three dogs with deficiency of the lysosomal enzymeα-l-iduronidase were treated by gene replacement therapy targeted at muscle. Direct intramuscular injections of plasmid encoding theα-l-iduronidase gene cDNA resulted in no detectable enzyme production, but may have resulted in immunologic sensitization to iduronidase protein, which the dogs lack totally. Myoblasts were grown from skeletal muscle biopsies and transduced with a retroviral vector containing the canine gene under control of the muscle creatine kinase enhancer. Several hundred-fold overexpression of enzyme production occurred in cultured cells; however, following reintroduction of the cultured cells into dogs, enzyme production declined rapidly. Concurrent with the falling enzyme levels, there was production of specific immunoglobulin G (IgG) antibody against iduronidase that was further associated with cellular infiltration of the myoblast injection sites. Most inflammatory cells were lymphocytes and plasma cells, suggesting local humoral and cellular immune responses to the enzyme-producing muscle cells. PCR analysis of tissues collected 2–22 weeks after the final treatment showed the persistence of Neo and canineα-l-iduronidase sequences in a progressively decreasing percentage of myoblasts. Results from this study in a canine model of mucopolysaccharidosis I underscore the fact that immunologic reactions to cells producing desirable, normal, but foreign, proteins may be as much an impediment to gene therapy as reactions to the viral vectors used to introduce the foreign gene.