OBJECTIVE: To explore the possibility of using the retrovirus vector (RV) in the gene therapy of neonatal neuropathy and the expression properties of target genes in the brain. METHODS: We combined human placenta alkaline phosphatase (hpAKP) cDNA obtained from plasmid DAP with RV pLXSN and transferred recombinant plasmid into packed cell line PT67 by the liposome entrapment method. We injected high titer recombinant hpAKP RV into the 1 day old mouse brain, and then determined the hpAKP expression in the brain by hybridation in situ and the enzyme histological chemistry method as well as the type of expression cells by the immunological histological chemistry method. RESULTS: The infected cells lay in the Ⅱ Ⅴ stratum of the cerebral cortex, the nuclei of the brain stem, the Purkenje cell stratum of the cerebellum, the vascular wall, and the cerebral mater. The area of positive cells increased from the 1st to the 7th day after the injection, but was smaller on the 28th day than on the 7th day after the injection, part of those being GFAP(-). RV was not found harmful to the nervous system. CONCLUSIONS: RV can transfer target genes into the newborn mouse brain effectively, including neurons. Target genes express normally. It is likely that RV could be used in the gene therapy of neonatal neuropathy."/>
重组hpAKP逆转录病毒载体的构建及其在新生小鼠脑内的表达
Abstract：OBJECTIVE: To explore the possibility of using the retrovirus vector (RV) in the gene therapy of neonatal neuropathy and the expression properties of target genes in the brain. METHODS: We combined human placenta alkaline phosphatase (hpAKP) cDNA obtained from plasmid DAP with RV pLXSN and transferred recombinant plasmid into packed cell line PT67 by the liposome entrapment method. We injected high titer recombinant hpAKP RV into the 1 day old mouse brain, and then determined the hpAKP expression in the brain by hybridation in situ and the enzyme histological chemistry method as well as the type of expression cells by the immunological histological chemistry method. RESULTS: The infected cells lay in the Ⅱ Ⅴ stratum of the cerebral cortex, the nuclei of the brain stem, the Purkenje cell stratum of the cerebellum, the vascular wall, and the cerebral mater. The area of positive cells increased from the 1st to the 7th day after the injection, but was smaller on the 28th day than on the 7th day after the injection, part of those being GFAP(-). RV was not found harmful to the nervous system. CONCLUSIONS: RV can transfer target genes into the newborn mouse brain effectively, including neurons. Target genes express normally. It is likely that RV could be used in the gene therapy of neonatal neuropathy.
