title: Gene transfer to the cerebellum.
abstract: There are several diseases for which gene transfer therapy to the cerebellum might be practicable. In these studies, we used recombinant Tag-deleted SV40-derived vectors (rSV40s) to study gene delivery targeting the cerebellum. These vectors transduce neurons and microglia very effectively in vitro and in vivo, and so we tested them to evaluate gene transfer to the cerebellum in vivo. Using a rSV40 vector carrying human immunodeficiency virus (HIV)-Nef with a C-terminal FLAG epitope, we characterized the distribution, duration, and cell types transduced. Rats received test and control vectors by stereotaxic injection into the cerebellum. Transgene expression was assessed 1, 2, and 4 weeks later by immunostaining of serial brain sections. FLAG epitope-expressing cells were seen, at all times after vector administration, principally detected in the Purkinje cells of the cerebellum, identified as immunopositive for calbindin. Occasional microglial cells were tranduced; transgene expression was not detected in astrocytes or oligodendrocytes. No inflammatory or other reaction was detected at any time. Thus, SV40-derived vectors can deliver effective, safe, and durable transgene expression to the cerebellum.
authors.affiliation: Department of Pathology, Jefferson Medical College, Thomas Jefferson University, 1020 Locust Street, Room 255, Philadelphia, PA 19107, USA. jplouboutin@hotmail.com
authors.forename: Jean-Pierre
authors.initials: JP
authors.lastname: Louboutin
authors.forename: Beverly A S
authors.initials: BA
authors.lastname: Reyes
authors.forename: Elisabeth J
authors.initials: EJ
authors.lastname: Van Bockstaele
authors.forename: David S
authors.initials: DS
authors.lastname: Strayer
mesh_terms: D000818:Animals; D002452:Cell Count / Q000379:methods; D002531:Cerebellum / Q000166:cytology / Q000378:metabolism*; D005260:Female; D015870:Gene Expression / Q000502:physiology; D015693:Gene Products, nef / Q000235:genetics / Q000378:metabolism; D005822:Genetic Vectors / Q000502:physiology; D009419:Nerve Tissue Proteins / Q000378:metabolism; D009474:Neurons / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D013539:Simian virus 40 / Q000235:genetics; D018709:Statistics, Nonparametric; D014161:Transduction, Genetic / Q000379:methods*
chemical_list: D015693:Gene Products, nef; D009419:Nerve Tissue Proteins