Tropism and toxicity of adeno-associated viral vector serotypes 1, 2, 5, 6, 7, 8, and 9 in rat neurons and glia in vitro
Tropism and toxicity of adeno-associated viral vector serotypes 1, 2, 5, 6, 7, 8, and 9 in rat neurons and glia in vitro
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DOI：
10.1016/j.virol.2007.10.007
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发表时间：
2008-03-01
期刊：
影响因子：
3.7
通讯作者：
Harvey, Brandon K.
中科院分区：
文献类型：
--
作者：
Howard, Douglas B.;Powers, Kathleen;Harvey, Brandon K.
重组腺相关病毒（rAAV）载体经常用于向中枢神经系统的基因递送，并且能够在体外转导神经元和神经胶质。在这项研究中，七个血清型的rAAV载体表达绿色荧光蛋白（GFP）的特点是在原代皮层细胞来源于胚胎大鼠脑的向性和毒性。在转导后2天，血清型1和5至8主要在神经胶质中表达GFP，但在转导后6天，除了AAV 5外，表达是神经元的。AAV 2和9产生最小GFP表达。使用细胞活力测定，对于除AAV 2和9之外的所有血清型，在较高的感染复数（MOI）下观察到毒性。AAV 1和5-8的毒性主要影响神经胶质，如神经胶质标志物免疫反应性的丧失所示。GFP基因中的移码突变降低了血清型1、5和6的总体毒性，但不降低血清型7和8的总体毒性，这表明毒性不仅仅是由于GFP的过表达。总的来说，在原代皮质培养物上的AAV血清型之间观察到不同的向性和毒性，具有总体优先的胶质细胞转导和毒性。爱思唯尔公司出版
Recombinant adeno-associated viral (rAAV) vectors are frequently used for gene delivery to the central nervous system and are capable of transducing neurons and glia in vitro. In this study, seven serotypes of a rAAV vector expressing green fluorescent protein (GFP) were characterized for tropism and toxicity in primary cortical cells derived from embryonic rat brain. At 2 days after transduction, serotypes 1 and 5 through 8 expressed GFP predominately in glia, but by 6 days post-transduction expression was neuronal except for AAV5. AAV2 and 9 produced minimal GFP expression. Using cell viability assays, toxicity was observed at higher multiplicities of infection (MOI) for all serotypes except AAV2 and 9. The toxicity of AAV1 and 5-8 affected mostly glia as indicated by a loss of glial-marker immunoreactivity. A frameshift mutation in the GFP gene reduced overall toxicity for serotypes 1, 5 and 6, but not 7 and 8 suggesting that the toxicity was not solely due to the overexpression of GFP. Collectively, a differential tropism and toxicity was observed among the AAV serotypes on primary cortical cultures with an overall preferential glial transduction and toxicity. Published by Elsevier Inc.