Transfer of the nerve growth factor gene into cell lines and cultured neurons using a defective herpes simplex virus vector. Transfer of the NGF gene into cells by a HSV-1 vector.
Nerve growth factor (NGF) can be expressed in cells by gene transfer using a defective Herpes Simplex virus type 1 (HSV-1) vector. In this report, the defective HSV-1 vector, pHSVngf, is used to infect established cell lines and cultured neurons. Infection of cell lines with pHSVngf results in gene transcription, correct RNA processing, and production of biologically active NGF. Infection of the PC12 neuronal cell line results in the production of biologically active NGF and infection of NGF-dependent neonatal sympathetic neurons in primary culture with pHSVngf leads to neuronal survival in the absence of exogenously-added NGF. NGF expressed by pHSVngf-infected cells does not appear to work through an autocrine intracellular pathway since NGF antibody added to culture media of infected cells could block NGF action. Infection with pHSVngf of cholinergic striatal or septal neurons in dissociated cell culture resulted in an increase in choline acetyltransferase activity. These studies demonstrate the efficacy of defective HSV-1 vectors for delivery and expression of neurotrophin genes in cultured neural cells.
Geschwind
M D
MD
Department of Medicine, Albert Einstein College of Medicine, Bronx, NY 10461.
Kessler
J A
JA
Geller
A I
AI
Federoff
H J
HJ
D016475:3T3 Cells; D000818:Animals; D001483:Base Sequence; D001921:Brain / Q000378:metabolism*; D002478:Cells, Cultured; D002795:Choline O-Acetyltransferase / Q000378:metabolism; D003342:Corpus Striatum / Q000378:metabolism; D017931:DNA Primers; D003673:Defective Viruses; D004622:Embryo, Mammalian; D005822:Genetic Vectors; D013006:Growth Hormone / Q000235:genetics; D018259:Herpesvirus 1, Human; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D009414:Nerve Growth Factors / Q000096:biosynthesis* / Q000235:genetics; D009474:Neurons / Q000378:metabolism*; D016716:PC12 Cells; D016133:Polymerase Chain Reaction; D011134:Polysaccharides / Q000096:biosynthesis* / Q000235:genetics; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis; D014162:Transfection / Q000379:methods*
D017931:DNA Primers; D009414:Nerve Growth Factors; D011134:Polysaccharides; D011994:Recombinant Proteins; C010491:neurotropin; D013006:Growth Hormone; D002795:Choline O-Acetyltransferase