Stone Daniel, Xiong Weidong, Williams Judith C, David Anne, Lowenstein Pedro R, Castro Maria G
Gene Therapeutics Research Institute, Cedars-Sinai Medical Center, Research Pavilion, Suite 5090, 8700 Beverly Boulevard, Los Angeles, California 90048, USA.
Mol Ther. 2003 Sep;8(3):400-11. doi: 10.1016/s1525-0016(03)00178-3.
Despite their ability to provide long-term transgene expression in the central nervous system of naïve hosts, the use of first-generation adenovirus (Ad) vectors for the treatment of chronic neurological disorders is limited by peripheral immunization, which stimulates anti-adenovirus immune responses and causes severe inflammation in the central nervous system (CNS) and elimination of transgene expression. The purpose of this study was to investigate the roles of NF-kappaB and interleukin-1 (IL-1) during inflammatory responses to Ads in the CNS of naïve and preimmunized rats. We assessed activation of macrophages/microglia, up-regulation of MHC I expression, infiltration of leukocytes, and transgene expression following delivery of Ads to the rat striatum. After delivery of increasing doses of adenoviral vectors expressing various anti-inflammatory agents (e.g., NF-kappaB or IL-1 inhibitors) to naïve rats, no reduction in Ad-mediated CNS inflammation was seen 1 week after delivery of Ads, compared to a control Ad.hCMV.beta-galactosidase (RAd.35) virus. We then assessed CNS inflammation and transgene expression at a time when control transgene expression would be completely eliminated, i.e., 1 month post-vector injection into the brain. This would optimize the assessment of an anti-inflammatory agent expressed by an adenoviral vector that could either delay or diminish immune system-mediated elimination of transgene expression. As expected, at 1 month postinfection, control preimmunized rats receiving Ad.mCMV.beta-galactosidase (RAd.36)/saline or RAd.36/Ad.null (RAd.0) showed complete elimination of beta-galactosidase expression in the brain and levels of inflammation comparable to those of naïve animals. However, animals injected with RAd.36 in combination with Ads expressing NF-kappaB or IL-1 inhibitors showed a delayed elimination of beta-galactosidase compared to controls. As predicted, the extended presence of transgene expression was accompanied by increased levels of CNS inflammation. This suggests that blocking NF-kappaB or IL-1 delays, albeit partially, transgene elimination in the presence of a preexisting systemic immune response. Prolonged transgene expression is predicted to extend concurrent brain inflammation, as noted earlier. Taken together these data demonstrate a role for NF-kappaB and IL-1 in immune system-mediated elimination of Ad-mediated CNS transgene expression.
尽管第一代腺病毒（Ad）载体能够在未经免疫的宿主中枢神经系统中提供长期转基因表达，但用于治疗慢性神经疾病时却受到外周免疫的限制，外周免疫会刺激抗腺病毒免疫反应，导致中枢神经系统（CNS）严重炎症并消除转基因表达。本研究的目的是调查在未经免疫和预先免疫的大鼠中枢神经系统中，核因子κB（NF-κB）和白细胞介素-1（IL-1）在对腺病毒的炎症反应中的作用。我们评估了将腺病毒递送至大鼠纹状体后巨噬细胞/小胶质细胞的激活、MHC I表达的上调、白细胞浸润以及转基因表达情况。向未经免疫的大鼠递送表达各种抗炎剂（如NF-κB或IL-1抑制剂）的腺病毒载体，剂量逐渐增加，在递送腺病毒1周后，与对照腺病毒hCMV.β-半乳糖苷酶（RAd.35）病毒相比，未观察到Ad介导的中枢神经系统炎症减轻。然后，我们在对照转基因表达将被完全消除的时间点，即向脑内注射载体1个月后，评估中枢神经系统炎症和转基因表达。这将优化对腺病毒载体表达的抗炎剂的评估，该抗炎剂可能延迟或减少免疫系统介导的转基因表达消除。正如预期的那样，在感染后1个月，接受Ad.mCMV.β-半乳糖苷酶（RAd.36）/生理盐水或RAd.36/Ad.null（RAd.0）的预先免疫的对照大鼠，脑内β-半乳糖苷酶表达完全消除，炎症水平与未经免疫的动物相当。然而，注射RAd.36与表达NF-κB或IL-1抑制剂的腺病毒组合的动物，与对照相比，β-半乳糖苷酶的消除延迟。正如所预测的，转基因表达的延长伴随着中枢神经系统炎症水平的增加。这表明在存在预先存在的全身免疫反应的情况下，阻断NF-κB或IL-1尽管只是部分地延迟了转基因的消除。如前所述，预计转基因表达的延长会使并发的脑部炎症延长。综合这些数据表明，NF-κB和IL-1在免疫系统介导的Ad介导的中枢神经系统转基因表达消除中发挥作用。
