Chen Juan, Guo Xiaojuan, Zou Xiaohui, Wang Min, Yang Chunlei, Hou Wenzhe, Sprindzuk Matvey V, Lu Zhuozhuang
NHC Key Laboratory of Medical Virology and Viral Diseases, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, China.
School of Public Health, Baotou Medical College, Inner Mongolia University of Science and Technology, Baotou 014040, China.
Viruses. 2024 Mar 31;16(4):550. doi: 10.3390/v16040550.
The administration route affects the biodistribution of a gene transfer vector and the expression of a transgene. A simian adenovirus 1 vector carrying firefly luciferase and GFP reporter genes (SAdV1-GFluc) were constructed, and its biodistribution was investigated in a mouse model by bioluminescence imaging and virus DNA tracking with real-time PCR. Luciferase activity and virus DNA were mainly found in the liver and spleen after the intravenous administration of SAdV1-GFluc. The results of flow cytometry illustrated that macrophages in the liver and spleen as well as hepatocytes were the target cells. Repeated inoculation was noneffective because of the stimulated serum neutralizing antibodies (NAbs) against SAdV-1. A transient, local expression of low-level luciferase was detected after intragastric administration, and the administration could be repeated without compromising the expression of the reporter gene. Intranasal administration led to a moderate, constant expression of a transgene in the whole respiratory tract and could be repeated one more time without a significant increase in the NAb titer. An immunohistochemistry assay showed that respiratory epithelial cells and macrophages in the lungs were transduced. High luciferase activity was restricted at the injection site and sustained for a week after intramuscular administration. A compromised transgene expression was observed after a repeated injection. When these mice were intramuscularly injected for a third time with the human adenovirus 5 (HAdV-5) vector carrying a luciferase gene, the luciferase activity recovered and reached the initial level, suggesting that the sequential use of SAdV-1 and HAdV-5 vectors was practicable. In short, the intranasal inoculation or intramuscular injection may be the preferred administration routes for the novel SAdV-1 vector in vaccine development.
给药途径会影响基因转移载体的生物分布以及转基因的表达。构建了携带萤火虫荧光素酶和绿色荧光蛋白报告基因的猿猴腺病毒1载体(SAdV1-GFluc),并通过生物发光成像和实时PCR病毒DNA追踪在小鼠模型中研究其生物分布。静脉注射SAdV1-GFluc后,荧光素酶活性和病毒DNA主要存在于肝脏和脾脏中。流式细胞术结果表明,肝脏、脾脏中的巨噬细胞以及肝细胞是靶细胞。由于刺激产生了针对SAdV-1的血清中和抗体(NAbs),重复接种无效。胃内给药后检测到低水平荧光素酶的短暂局部表达,并且可以重复给药而不影响报告基因的表达。鼻内给药导致转基因在整个呼吸道中适度、持续表达,并且可以再重复一次而不会使NAb滴度显著增加。免疫组织化学分析表明,肺中的呼吸道上皮细胞和巨噬细胞被转导。肌肉注射后,高荧光素酶活性局限于注射部位并持续一周。重复注射后观察到转基因表达受损。当这些小鼠第三次肌肉注射携带荧光素酶基因的人腺病毒5(HAdV-5)载体时,荧光素酶活性恢复并达到初始水平,这表明依次使用SAdV-1和HAdV-5载体是可行的。简而言之,鼻内接种或肌肉注射可能是新型SAdV-1载体在疫苗开发中的首选给药途径。