Systemic FIV vector administration: transduction of CNS immune cells and Purkinje neurons.
The systemic effects of gene therapy have been previously described in a variety of peripheral organs following intravenous administration or intraperitoneal inoculation of viral vectors, as well as in the brain following intracranial administration. However, limited information is available on the ability of viral vectors to cross the blood-brain barrier and infect cells located within the central nervous system (CNS). We employed a VSV-G pseudotyped FIV(lacZ) vector capable of transducing dividing, growth-arrested, as well as post-mitotic cells with the reporter gene lacZ. Adult mice were injected intraperitoneally with FIV(lacZ), and the expression of beta-galactosidase was studied 5 weeks following treatment in the brain, liver, spleen and kidney by X-gal histochemistry and immunocytochemistry. Interestingly, relatively low doses of FIV(lacZ) administered intraperitoneally lead to beta-galactosidase detection in the brain and cerebellum. The identity of these cells was confirmed by double immunofluorescence, and included CD31-, CD3- and CD11b-positive cells. Fluorescent microspheres co-injected with FIV(lacZ) virus were identified within mononuclear cells in the brain parenchyma, suggesting infiltration of peripheral immune cells in the CNS. Cerebellar Purkinje neurons were also transduced in all adult-injected mice. Our observations indicate that relatively low doses of FIV(lacZ) administered intraperitoneally resulted in the transduction of immune cells in the brain, as well as a specific subset of cerebellar neurons.
Kyrkanides
Stephanos
S
Department of Dentistry, School of Medicine and Dentistry, University of Rochester, Rochester, NY, USA. stephanos_Kyrkanides@urmc.rochester.edu
Miller
Jennie H
JH
Federoff
Howard J
HJ
D000818:Animals; D000954:Antigens, Surface / Q000276:immunology; D001812:Blood-Brain Barrier / Q000821:virology; D001921:Brain / Q000166:cytology / Q000276:immunology / Q000821:virology*; D002634:Chemotaxis, Leukocyte / Q000235:genetics; D051546:Cyclooxygenase 2; D017930:Genes, Reporter / Q000235:genetics; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism*; D016089:Immunodeficiency Virus, Feline / Q000235:genetics*; D007274:Injections, Intraperitoneal; D007527:Isoenzymes / Q000378:metabolism; D007763:Lac Operon / Q000235:genetics; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000378:metabolism / Q000821:virology*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D011451:Prostaglandin-Endoperoxide Synthases / Q000378:metabolism; D011689:Purkinje Cells / Q000166:cytology / Q000378:metabolism / Q000821:virology*; D014161:Transduction, Genetic / Q000379:methods*; D019010:Vascular Cell Adhesion Molecule-1 / Q000378:metabolism; D001616:beta-Galactosidase / Q000235:genetics
D000954:Antigens, Surface; D007527:Isoenzymes; D019010:Vascular Cell Adhesion Molecule-1; D051546:Cyclooxygenase 2; D011451:Prostaglandin-Endoperoxide Synthases; D001616:beta-Galactosidase