中华医学杂志（英文版）Issue(9)：1370-1376,7.
Expression of human nerve growth factor β gene in central nervous system mediated by recombinant adeno-associated viruses type-2 vector
Expression of human nerve growth factor β gene in central nervous system mediated by recombinant adeno-associated viruses type-2 vector
高凯 1吴勇杰 2吴小兵 3饶春明 1王军志1
作者信息
- 1. National Institute for the Control of Pharmaceutical and Biological Products, Beijing 100050, China
- 2. Department of Pharmacology, Lanzhou Medical University, Lanzhou 730000, China
- 3. National 863 R&D Base of Viral Gene Vector, Beijing 100176, China
- 折叠
摘要
Abstract
Background Neurone atrophy and loss are major causes of chronic neurodegenerative disorders such as Alzheimer's disease. Despite many pharmacotherapies for neurodegeneration, there are no accepted treatments. We investigated the feasibility of human nerve growth factor β (hNGFβ) gene expression mediated by recombinant adeno-associated viruses type-2 (rAAV-2) vector in the central nervous system (CNS) after blood brain barrier (BBB) disruption.Methods rAAV-2 containing hNGFβ gene was constructed. The ability of hNGFβ gene mediated by rAAV-2 vector (rAAV-2/hNGFβ) to transfect cells in vitro was confirmed by both ELISA and bioassay of hNGFβ in the culture supernatant of BHK-21 cells infected by rAAV-2/hNGFβ. rAAV-2/hNGFβ and rAAV-2/green fluorescence protein (GFP) were administrated separately to rat brains through internal carotid intubation after BBB disruption with hypertonic mannitol. Brain hNGFβ concentration was measured by ELISA and GFP in brain sections was examined by laser scan confocal microscope.Results After 48 hours, hNGFβ content in supernatant was up to (188.0±28.6) pg/ml when BHK-21 cells were infected by rAAV-2/hNGFβ at multiplicity of infection (MOI)1.0×106 vector genome. Neurone fibre outgrowths were obvious in dorsal root ganglion neurone assays by adding serum free culture medium harvested from BHK-21 cells exposed to rAAV-2/hNGFβ. Whole brain hNGFβ content in rAAV-2/hNGFβ transferred group was up to (636.2±140.6) pg/ml. hNGFβ content of BBB disruption in rAAV-2/hNGFβ infused group increased significantly compared to the control group (P<0.05). GFP expression was clearly observed in brain sections of rAAV-2/GFP transferred group.Conclusion rAAV-2/hNGFβ successfully expresses in the CNS after BBB disruption induced by hypertonic mannitol.关键词
human nerve growth factor β/adeno-associated virus/gene therapy/blood brain barrierKey words
human nerve growth factor β/adeno-associated virus/gene therapy/blood brain barrier分类
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高凯,吴勇杰,吴小兵,饶春明,王军志..Expression of human nerve growth factor β gene in central nervous system mediated by recombinant adeno-associated viruses type-2 vector[J].中华医学杂志（英文版）,2004,(9):1370-1376,7.
