Jani A, Menichella D, Jiang H, Chbihi T, Acsadi G, Kamholz J, Shy M E
Department of Neurology, Wayne State University School of Medicine, Detroit, Michigan 48201, USA.
Ann N Y Acad Sci. 1999 Sep 14;883:397-414.
In a previous report, we demonstrated that a first generation (E1- and E3-deleted) recombinant adenovirus can transduce expression of the E. coli lacZ gene into Schwann cells, both in vitro and in vivo, suggesting that this method might be useful for future therapy of peripheral neuropathy, including CMT1. Adenoviral-mediated gene transfer was limited, however, by demyelination and Wallerian degeneration at the site of virus injection, as well as by attenuation of viral gene expression over time. In our current work we have optimized adenoviral-mediated gene expression after intraneural injection into sciatic nerve. Using an improved injection protocol, peak expression of lacZ occurs between 10 and 14 days after injection of 2-week-old animals, decreases thereafter, and there is minimal associated tissue injury. In contrast, very few adenoviral-infected Schwann cells are found in nerves of adult animals 10 days after injection, probably due to immune clearance of viral-infected cells. Consistent with this notion, high levels of lacZ are found in sciatic nerve 30 days after injection of adult SCOD mice, which have a genetic defect in both cellular and humoral immunity, of adult beta 2 microglobulin-deficient mice (beta 2 M-/-), which have a genetic defect in cellular immunity, or of adult mice treated with the immunosuppressing agent FK506. In addition, adenoviral-infected Schwann cells co-cultured with axons in vitro, in the absence of a host immune response, ensheath axons and express lacZ for at least 8 weeks. These data thus demonstrate that expression of first generation recombinant adenovirus in sciatic nerve in adult mice, as in other tissues, is limited mainly by the host cellular immune response to the virus, which can be overcome by attenuation of host cell-mediated immunity. Adenoviral vectors might thus be used to modulate Schwann cell gene expression in patients with peripheral neuropathy after appropriate immunosuppression.
在之前的一份报告中，我们证明了第一代（E1和E3缺失）重组腺病毒能够在体外和体内将大肠杆菌lacZ基因的表达转导至雪旺细胞，这表明该方法可能对包括CMT1在内的周围神经病变的未来治疗有用。然而，腺病毒介导的基因转移受到病毒注射部位的脱髓鞘和华勒变性以及病毒基因表达随时间衰减的限制。在我们当前的工作中，我们优化了将腺病毒介导的基因注射到坐骨神经后的表达。使用改进的注射方案，在对2周龄动物注射后10至14天出现lacZ的峰值表达，此后下降，并且相关的组织损伤最小。相比之下，在注射后10天的成年动物神经中发现极少的腺病毒感染的雪旺细胞，这可能是由于病毒感染细胞的免疫清除。与此观点一致，在注射成年SCOD小鼠（其在细胞免疫和体液免疫方面均有遗传缺陷）、成年β2微球蛋白缺陷小鼠（β2M-/-，其在细胞免疫方面有遗传缺陷）或用免疫抑制剂FK506处理的成年小鼠30天后，在坐骨神经中发现高水平的lacZ。此外，在体外与轴突共培养的腺病毒感染的雪旺细胞，在没有宿主免疫反应的情况下，包裹轴突并表达lacZ至少8周。因此，这些数据表明，与其他组织一样，成年小鼠坐骨神经中第一代重组腺病毒的表达主要受宿主对病毒的细胞免疫反应限制，而这种限制可通过减弱宿主细胞介导的免疫来克服。因此，在适当的免疫抑制后，腺病毒载体可用于调节周围神经病变患者的雪旺细胞基因表达。
