In vivo gene transfer using a nonprimate lentiviral vector pseudotyped with Ross River Virus glycoproteins.
Vectors derived from lentiviruses provide a promising gene delivery system. We examined the in vivo gene transfer efficiency and tissue or cell tropism of a feline immunodeficiency virus (FIV)-based lentiviral vector pseudotyped with the glycoproteins from Ross River Virus (RRV). RRV glycoproteins were efficiently incorporated into FIV virions, generating preparations of FIV vector, which after concentration attain titers up to 1.5 x 10(8) TU/ml. After systemic administration, RRV-pseudotyped FIV vectors (RRV/FIV) predominantly transduced the liver of recipient mice. Transduction efficiency in the liver with the RRV/FIV was ca. 20-fold higher than that achieved with the vesicular stomatitis virus G protein (VSV-G) pseudotype. Moreover, in comparison to VSV-G, the RRV glycoproteins caused less cytotoxicity, as determined from the levels of glutamic pyruvic transaminase and glutamic oxalacetic transaminase in serum. Although hepatocytes were the main liver cell type transduced, nonhepatocytes (mainly Kupffer cells) were also transduced. The percentages of the transduced nonhepatocytes were comparable between RRV and VSV-G pseudotypes and did not correlate with the production of antibody against the transgene product. After injection into brain, RRV/FIV preferentially transduced neuroglial cells (astrocytes and oligodendrocytes). In contrast to the VSV-G protein that targets predominantly neurons, <10% of the brain cells transduced with the RRV pseudotyped vector were neurons. Finally, the gene transfer efficiencies of RRV/FIV after direct application to skeletal muscle or airway were also examined and, although transgene-expressing cells were detected, their proportions were low. Our data support the utility of RRV glycoprotein-pseudotyped FIV lentiviral vectors for hepatocyte- and neuroglia-related disease applications.
Kang
Yubin
Y
Program in Gene Therapy, Department of Pediatrics, University of Iowa College of Medicine, Iowa City, Iowa 52242, USA.
Stein
Colleen S
CS
Heth
Jason A
JA
Sinn
Patrick L
PL
Penisten
Andrea K
AK
Staber
Patrick D
PD
Ratliff
Kenneth L
KL
Shen
Hong
H
Barker
Carrie K
CK
Martins
Inês
I
Sharkey
C Matthew
CM
Sanders
David Avram
DA
McCray
Paul B
PB
Davidson
Beverly L
BL
D000818:Animals; D002213:Capsid / Q000235:genetics*; D036022:Capsid Proteins; D002415:Cats; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D022781:Hepatocytes / Q000378:metabolism / Q000821:virology; D016089:Immunodeficiency Virus, Feline / Q000235:genetics* / Q000378:metabolism; D008099:Liver / Q000166:cytology; D008562:Membrane Glycoproteins / Q000235:genetics*; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D018482:Muscle, Skeletal / Q000378:metabolism / Q000821:virology; D009457:Neuroglia / Q000378:metabolism / Q000821:virology; D012398:Ross River virus / Q000235:genetics* / Q000378:metabolism; D014161:Transduction, Genetic; D019076:Transgenes / Q000235:genetics / Q000502:physiology; D014759:Viral Envelope Proteins / Q000235:genetics*; D014771:Virion / Q000235:genetics / Q000378:metabolism; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
D036022:Capsid Proteins; C110266:E1 protein, Ross River virus; C085817:E2 protein, Ross River virus; D008562:Membrane Glycoproteins; D014759:Viral Envelope Proteins; D001616:beta-Galactosidase