Kajiwara K, Byrnes A P, Charlton H M, Wood M J, Wood K J
Nuffield Department of Surgery, University of Oxford, Johm Radcliffe Hospital, UK.
Hum Gene Ther. 1997 Feb 10;8(3):253-65. doi: 10.1089/hum.1997.8.3-253.
We have investigated the immune response to E1-deleted adenovirus vectors encoding the lacZ gene introduced into the brains of adult mice. Injection of these nonreplicating vectors caused a marked inflammatory response in the brain as assessed by immunocytochemistry and flow cytometry of leukocytes. Infiltrating leukocytes were detectable within 2 days of injection and reached a maximum by 9 days. Thereafter, the number of infiltrating cells decreased, but a small number persisted in the brain until day 60. Between 2 and 4 days after injection, the percentage of CD8+ cells detectable increased whereas the percentage of CD4+ cells present in the infiltrating population did not significantly increase until day 6, peaking on day 15. Activated CD25+ T cells were detectable between days 6 and 15. beta-Galactosidase (beta-Gal), the product of the lacZ gene encoded by the vector, was also detected, both at the injection site in the striatum and also in the substantia nigra. Expression peaked between 4 and 6 days but a small number of beta-Gal+ cells was still seen at 60 days after injection. This study demonstrates that a quantitative analysis of the immune responses caused by a nonreplicating adenovirus vector is possible in the brain. E1-deleted adenoviral vectors trigger a strong inflammatory response in the brain, but this immune response is not sufficient to eliminate completely expression of genes encoded by the adenoviral construct.
我们研究了成年小鼠脑内引入编码lacZ基因的E1缺失腺病毒载体后的免疫反应。通过白细胞免疫细胞化学和流式细胞术评估，注射这些非复制型载体可在脑内引发明显的炎症反应。注射后2天内可检测到浸润的白细胞，9天时达到峰值。此后，浸润细胞数量减少，但少量细胞在脑内持续存在至第60天。注射后2至4天，可检测到的CD8 +细胞百分比增加，而浸润群体中CD4 +细胞百分比直到第6天才显著增加，在第15天达到峰值。在第6天至15天之间可检测到活化的CD25 + T细胞。载体编码的lacZ基因产物β-半乳糖苷酶（β-Gal）在纹状体注射部位和黑质中均有检测到。表达在4至6天达到峰值，但注射后60天时仍可见少量β-Gal +细胞。本研究表明，在脑内对非复制型腺病毒载体引起的免疫反应进行定量分析是可行的。E1缺失腺病毒载体在脑内引发强烈的炎症反应，但这种免疫反应不足以完全消除腺病毒构建体编码基因的表达。
