Helper-free HSV-1 amplicons elicit a markedly less robust innate immune response in the CNS.
Helper-free HSV-1 amplicons elicit a markedly less robust innate immune response in the CNS.
复制标题
无辅助 HSV-1 扩增子在中枢神经系统中引发的先天免疫反应明显较差。
DOI：
10.1016/s1525-0016(02)00036-9
复制
发表时间：
2003
期刊：
影响因子：
--
通讯作者：
Federoff,HowardJ
中科院分区：
文献类型：
--
作者：
Olschowka,JohnA;Bowers,WilliamJ;Hurley,SeanD;Mastrangelo,MichaelA;Federoff,HowardJ
为了研究和治疗慢性中枢神经系统疾病，开发和实施直接基因转移技术本身就需要考虑媒介的安全性。基于病毒的载体是最有效的方式，但在体内给药时具有诱导强大的先天和获得性免疫反应的潜力，这些反应可能是由于病毒颗粒成分、由此产生的病毒基因表达和/或转基因表达引起的。在目前的研究中，我们描述了在基于单纯疱疹病毒1型的扩增载体的立体定向递送时所引起的先天反应。给C57BL/6小鼠注射无菌生理盐水，用常规辅助病毒技术包装的β-半乳糖苷酶表达扩增子(HSVlac)或无辅助病毒的HSVlac。在转导后1天或5天处死小鼠，通过免疫细胞化学和定量RT-PCR分析各种趋化因子、细胞因子和黏附分子基因的转录。所有的注射都引起了炎症，第一天血/脑屏障开放，与生理盐水对照组相比，两种扩增子制剂都增强了这种开放。到第5天，致炎细胞因子(IL-1β、肿瘤坏死因子-α、干扰素-γ)、趋化因子(单核细胞趋化蛋白-1、IP-10)和黏附分子(ICAM-1)的基因表达水平在注射生理盐水的小鼠中已恢复到基线水平，而在辅助病毒无扩增子组则恢复到接近基线水平。相比之下，注射辅助病毒包装扩增子的小鼠即使在第5天也能诱导炎症分子的表达和免疫细胞的渗透。综上所述，我们证明了无辅助病毒扩增子制剂表现出更安全的先天免疫反应，这可能是由于辅助病毒基因表达缺失的结果，并为未来基于扩增子的中枢神经系统基因转移策略提供支持。
The development and implementation of direct gene transfer technologies for the study and treatment of chronic CNS disorders inherently requires consideration of vector safety. Virus-based vectors represent the most efficient modalities but harbor the potential to induce vigorous innate and adaptive immune responses when administeredin vivo.These responses can arise because of virus particle components, resultant viral gene expression, and/or transgene expression. In the current study, we describe the innate responses elicited upon stereotactic delivery of herpes simplex virus type 1-based amplicon vectors. C57BL/6 mice were injected with sterile saline, β-galactosidase-expressing amplicon (HSVlac) packaged by a conventional helper virus-based technique, or helper virus-free HSVlac. After killing the mice at either 1 or 5 days after transduction, we analyzed them by immunocytochemistry and quantitative RT-PCR for various chemokine, cytokine, and adhesion molecule gene transcripts. All injections induced inflammation, with blood/brain barrier opening on day 1 that was enhanced with both amplicon preparations as compared with saline controls. By day 5, mRNA levels for the pro-inflammatory cytokines (IL-1β, TNF-α, IFN-γ), chemokines (MCP-1, IP-10), and an adhesion molecule (ICAM-1) had returned to baseline in saline-injected mice and to near-baseline levels in helper virus-free amplicon groups. In contrast, mice injected with helper virus-packaged amplicon stocks elicited elevated inflammatory molecule expression and immune cell infiltration even at day 5. In aggregate, we demonstrate that helper virus-free amplicon preparations exhibit a safer innate immune response profile, presumably as a result of the absence of helper virus gene expression, and provide support for future amplicon-based CNS gene transfer strategies.