Characterization of lentiviral vector-mediated gene transfer in adult mouse brain.
Lentiviral vectors are promising tools for gene transfer into the central nervous system. We have characterized in detail transduction with human immunodeficiency virus type 1 (HIV-1)-derived vectors encoding enhanced green fluorescent protein (eGFP) in the adult mouse brain. Different brain regions such as the striatum, hippocampus, and the lateral ventricle were targeted. The eGFP protein was transported anterogradely in the nigrostriatal pathway, but we have found no evidence of transport of the lentiviral vector particle. The performance levels of the different generations of packaging and transfer plasmid were compared. Omission of the accessory genes from the packaging plasmid resulted in a modest decrease in transgene expression. Inclusion of the woodchuck hepatitis posttranscriptional regulatory element, on the one hand, and the central polypurine tract and termination sequences, on the other hand, in the transfer vector each resulted in a 4- to 5-fold increase in transgene expression levels. Combination of both elements enhanced expression levels more than the sum of the individual components, suggesting a synergistic effect. In the serum of mice injected with lentiviral vectors a humoral response to vector proteins was detected, but this did not compromise transgene expression. Immune response to the transgene was found only in a minority of the animals.
Baekelandt
Veerle
V
Gene Therapy Program, Laboratory for Experimental Neurosurgery and Neuroanatomy, Katholieke Universiteit Leuven, Provisorium 1, Minderbroedersstraat 17, B-3000 Leuven, Belgium. veerle.baekelandt@med.keuleuven.ac.be
Claeys
Anje
A
Eggermont
Kristel
K
Lauwers
Erwin
E
De Strooper
Bart
B
Nuttin
Bart
B
Debyser
Zeger
Z
D000818:Animals; D017209:Apoptosis; D015153:Blotting, Western; D001921:Brain / Q000378:metabolism* / Q000473:pathology; D002455:Cell Division; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D017919:Hepatitis B Virus, Woodchuck / Q000235:genetics; D006624:Hippocampus / Q000378:metabolism; D016086:Lentivirus / Q000235:genetics*; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D016513:Mice, SCID; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D009457:Neuroglia / Q000473:pathology; D009474:Neurons / Q000473:pathology; D010957:Plasmids / Q000378:metabolism; D013997:Time Factors; D014158:Transcription, Genetic; D019076:Transgenes
D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins