Stability of lentiviral vector-mediated transgene expression in the brain in the presence of systemic antivector immune responses
Stability of lentiviral vector-mediated transgene expression in the brain in the presence of systemic antivector immune responses
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DOI：
10.1089/hum.2005.16.741
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发表时间：
2005-06-01
影响因子：
4.2
通讯作者：
Lowenstein, PR
中科院分区：
文献类型：
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作者：
Abordo-Adesida, E;Follenzi, A;Lowenstein, PR
慢病毒载体是中枢神经系统基因治疗的有前途的工具。因此，重要的是表征它们与CNS中免疫系统的相互作用。这项工作的特点转基因表达和脑炎症的存在或不存在的免疫反应后产生的全身免疫慢病毒载体。我们表征了在CNS中用SIN-LV载体的转导。使用SIN-LV-GFP的剂量-反应曲线表明，在慢病毒载体转导单位的剂量为10（2）时，纹状体中可检测到转基因表达，在10（6）时表达最大，在注射的载体的最低和最高剂量之间炎性标志物的增加最小。我们的研究表明，将慢病毒载体注射到CNS中不会引起可测量的炎症反应。CNS注射后的全身免疫，与慢病毒载体表达相同的转基因作为载体注射到CNS中，引起CNS中的转基因表达的减少，伴随着炎性细胞浸润到CNS实质中的注射部位。然而，用携带不同转基因的慢病毒载体进行外周免疫不会减少转基因表达，也不会引起CNS炎症。将慢病毒载体注射到CNS中之前的全身免疫确定了预先存在的抗慢病毒免疫，无论转基因如何，都不影响转基因表达。此外，我们表明，转基因，而不是病毒粒子或载体成分，是负责提供抗原表位激活的免疫系统，对系统免疫与慢病毒。低免疫原性和延长转基因表达的存在下，预先存在的慢病毒免疫是令人鼓舞的数据，为未来使用慢病毒载体在中枢神经系统基因治疗。总之，测试的慢病毒载体诱导了不可检测的先天免疫应答的激活，并且仅当免疫应答针对转基因时，刺激针对慢病毒载体的适应性免疫应答才有效地引起转基因表达的降低。单独针对载体组分的全身性免疫应答不会引起脑炎症，这可能是因为载体衍生的表位未在CNS中呈递。
Lentiviral vectors are promising tools for gene therapy in the CNS. It is therefore important to characterize their interactions with the immune system in the CNS. This work characterizes transgene expression and brain inflammation in the presence or absence of immune responses generated after systemic immunization with lentiviral vectors. We characterized transduction with SIN-LV vectors in the CNS. A dose-response curve using SIN-LV-GFP demonstrated detectable transgene expression in the striatum at a dose of 10(2), and maximum expression at 10(6), transducing units of lentiviral vector, with minimal increase in inflammatory markers between the lowest and highest dose of vector injected. Our studies demonstrate that injection of a lentiviral vector into the CNS did not cause a measurable inflammatory response. Systemic immunization after CNS injection, with the lentiviral vector expressing the same transgene as a vector injected into the CNS, caused a decrease in transgene expression in the CNS, concomitantly with an infiltration of inflammatory cells into the CNS parenchyma at the injection site. However, peripheral immunization with a lentiviral vector carrying a different transgene did not diminish transgene expression, or cause CNS inflammation. Systemic immunization preceding injection of lentiviral vectors into the CNS determined that preexisting antilentiviral immunity, regardless of the transgene, did not affect transgene expression. Furthermore, we showed that the transgene, but not the virion or vector components, is responsible for providing antigenic epitopes to the activated immune system, on systemic immunization with lentivirus. Low immunogenicity and prolonged transgene expression in the presence of preexisting lentiviral immunity are encouraging data for the future use of lentiviral vectors in CNS gene therapy. In summary, the lentiviral vectors tested induced undetectable activation of innate immune responses, and stimulation of adaptive immune responses against lentiviral vectors was effective in causing a decrease in transgene expression only if the immune response was directed against the transgene. A systemic immune response against vector components alone did not cause brain inflammation, possibly because vector-derived epitopes were not being presented in the CNS.