Impairment of force generation after adenovirus-mediated gene transfer to muscle is alleviated by adenoviral gene inactivation and host CD8+ T cell deficiency.-猫眼课题宝

腺病毒介导的基因转移到肌肉后产生的力的损伤可以通过腺病毒基因失活和宿主 CD8 T 细胞缺陷得到缓解。

DOI：

发表时间：

1996

期刊：

Human Gene Therapy

影响因子：

4.2

通讯作者：

中科院分区：

医学2区

文献类型：

作者：

B. Petrof;Hanns Lochmüller;B. Massie;L. Yang;C. Macmillan;J. Zhao;J. Nalbantoglu;G. Karpati

重组腺病毒载体（AdV）有望作为一种手段，提供治疗基因的肌肉疾病，如杜氏肌营养不良症（DMD）。然而，我们以前已经表明，使用腺病毒是阻碍了发展减少的力量产生能力，这发生在1周内，并进行到至少1个月后，腺病毒交付免疫能力的动物。肌力产生的测定提供了一个敏感的和临床上重要的措施，对肌肉细胞功能的腺病毒介导的基因转移的潜在不利影响。在本研究中，我们研究了AdV相关基因表达和宿主T淋巴细胞反应在肌肉注射AdV后肌肉功能障碍发生中的作用。我们报告说，紫外线照射的AdV颗粒，这减少了AdV的转录活性，而不损害感染性（证实了原位聚合酶链反应），显着逆转早期（注射后4天）AdV诱导的收缩功能障碍的免疫小鼠以及缺乏有效的CD 8 + T细胞活性的小鼠。在免疫活性小鼠中，通常在AdV递送后4至30天发现力产生能力的叠加额外降低，沿着相关的转基因（β-半乳糖苷酶）表达丧失，这在很大程度上被不存在完整的CD 8 + T淋巴细胞应答所消除。此外，短期施用针对CD 4 + T细胞的中和抗体显著延长转基因表达，并显示出缓解AdV诱导的力产生能力降低的趋势。在CD 8+或CD 4 + T细胞缺乏的情况下，针对载体和转基因产物的细胞浸润和体液免疫应答也不同程度地减弱。我们的结论是，腺病毒相关基因的表达有一个早期的负面影响（可能是有毒的）的肌肉细胞功能，是独立的CD 8 + T细胞介导的免疫。相反，收缩障碍的进一步进展和伴随的AdV注射肌肉转基因表达的丧失在很大程度上取决于CD 8 + T细胞的活性。这些结果对未来一代载体的设计和AdV介导的基因转移到肌肉后免疫抑制治疗的潜在需求具有影响。

Recombinant adenovirus vectors (AdV) hold promise as a means of delivering therapeutic genes to muscle in diseases such as Duchenne muscular dystrophy (DMD). However, we have previously shown that the use of AdV is hampered by the development of reduced force-generating capacity, which occurs within 1 week and is progressive up to at least 1 month after AdV delivery in immune-competent animals. Determinations of muscle force production provide a sensitive and clinically important measure of potential adverse effects of AdV-mediated gene transfer on muscle cell function. In the present study, we investigated the role of AdV-related gene expression and host T lymphocyte responses in the genesis of muscle dysfunction following AdV injection of muscle. We report that UV-irradiation of AdV particles, which reduced AdV transcriptional activity without impairing infectivity (as confirmed by in situ polymerase chain reaction), significantly reversed early (4 days post-injection) AdV-induced contractile impairment in immune-competent mice as well as in mice lacking effective CD8+ T cell activity. The superimposed additional reduction in force-generating capacity normally found between 4 and 30 days post-AdV delivery in immune-competent mice, along with the associated loss of transgene (beta-galactosidase) expression, was largely abrogated by the absence of an intact CD8+ T lymphocyte response. Furthermore, short-term administration of a neutralizing antibody against CD4+ T cells significantly prolonged transgene expression and showed a trend toward mitigation of AdV-induced reductions in force-generating capacity. Cellular infiltration and humoral immune responses against the vector and transgene product were also blunted to varying degrees in the setting of CD8+ or CD4+ T cell deficiency. We conclude that AdV-related gene expression has an early negative (probably toxic) effect on muscle cell function that is independent of CD8+ T cell-mediated immunity. In contrast, further progression of contractile impairment and the accompanying loss of transgene expression from AdV-injected muscle are largely dependent upon the activity of CD8+ T cells. These results have implications for the design of future generation vectors and the potential need for immunosuppressive therapy after AdV-mediated gene transfer to muscle.