Immune responses to adeno-associated virus type 2 encoding channelrhodopsin-2 in a genetically blind rat model for gene therapy
Immune responses to adeno-associated virus type 2 encoding channelrhodopsin-2 in a genetically blind rat model for gene therapy
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DOI：
10.1038/gt.2010.140
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发表时间：
2011-03-01
期刊：
影响因子：
5.1
通讯作者：
Tomita, H.
中科院分区：
文献类型：
--
作者：
Sugano, E.;Isago, H.;Tomita, H.
我们以前曾报道过，将通道视紫红质-2（ChR 2）基因转导到视网膜神经节细胞中可以恢复遗传性失明、营养不良的皇家外科学院（RCS）大鼠的视觉功能。本研究试图揭示外源ChR 2基因表达的安全性和影响。通过玻璃体内注射向营养不良的RCS大鼠施用编码与Venus融合的ChR 2（rAAV-ChR 2 V）的腺相关病毒（AAV）2型。然而，在某些情况下，在非靶器官（肠、肺和心脏）中检测到rAAV-ChR 2基因表达。通过记录视觉诱发电位监测的ChR 2功能在整个观察期（64周）内保持稳定。未观察到视网膜组织学变化，也未观察到视网膜血管中白细胞粘附的炎症标记物。虽然检测到rAAV（0.01-12.21 μ g ml（-1））和ChR 2（0-4.77 μ g ml（-1））的抗体，但它们的水平太低，不可能被排斥。T淋巴细胞分析显示，T细胞的识别和短暂的炎症样免疫反应，直到1个月后的rAAV-ChR 2 V注射。总之，来源于莱茵衣藻的ChR 2可以在体内表达而不会产生免疫有害反应。这些发现将有助于研究ChR 2基因转移，以恢复视力的进展视网膜色素变性。Gene Therapy（2011）18，266-274; doi：10.1038/gt.2010.140; 2010年10月28日在线发表
We had previously reported that transduction of the channelrhodopsin-2 (ChR2) gene into retinal ganglion cells restores visual function in genetically blind, dystrophic Royal College of Surgeons (RCS) rats. In this study, we attempted to reveal the safety and influence of exogenous ChR2 gene expression. Adeno-associated virus (AAV) type 2 encoding ChR2 fused to Venus (rAAV-ChR2V) was administered by intra-vitreous injection to dystrophic RCS rats. However, rAAV-ChR2 gene expression was detected in non-target organs (intestine, lung and heart) in some cases. ChR2 function, monitored by recording visually evoked potentials, was stable across the observation period (64 weeks). No change in retinal histology and no inflammatory marker of leucocyte adhesion in the retinal vasculature were observed. Although antibodies to rAAV (0.01-12.21 mu g ml(-1)) and ChR2 (0-4.77 mu g ml(-1)) were detected, their levels were too low for rejection. T-lymphocyte analysis revealed recognition by T cells and a transient inflammation-like immune reaction only until 1 month after the rAAV-ChR2V injection. In conclusion, ChR2, which originates from Chlamydomonas reinhardtii, can be expressed without immunologically harmful reactions in vivo. These findings will help studies of ChR2 gene transfer to restore vision in progressed retinitis pigmentosa. Gene Therapy (2011) 18, 266-274; doi:10.1038/gt.2010.140; published online 28 October 2010