HIV-1-derived lentiviral vectors and fetal route of administration on transgene biodistribution and expression in rhesus monkeys.
The gene transfer efficiency of lentiviral vectors pseudotyped with vesicular stomatitis virus-glycoprotein (VSV-G) driven by the MND or CMV promoters and expressing the enhanced green fluorescent protein (EGFP) was investigated in fetal rhesus monkeys (Macaca mulatta) (N=21). Fetuses (50+/-10 days gestation; term 165+/-10 days) were injected under ultrasound guidance using an intraperitoneal (i.p.) or intrahepatic (i.h.) approach with a range of 1 x 10(7)-2 x 10(8) infectious particles/fetus. Analysis of transgene biodistribution and expression was performed in multiple tissues at 3-7 months postgene delivery using quantitative techniques. Overall, results indicated the following: (1) i.p. gene transfer at 40 days gestation resulted in a more diffuse distribution of the vector compared to administration at 60 days gestation; (2) vector biodistribution was similar after administration by the i.p. or i.h. routes; and (3) gene expression analysis in transduced tissues showed the presence of mRNA transcripts that correlated with the level of gene transfer. These studies suggest that fetal gene transfer using the i.p. and i.h. routes results in prolonged transduction and expression of the transgene in multiple tissues.
Jimenez
D F
DF
California National Primate Research Center, University of California, Davis, CA 95616-8542, USA.
Lee
C I
CI
O'Shea
C E
CE
Kohn
D B
DB
Tarantal
A F
AF
D000818:Animals; D001853:Bone Marrow / Q000378:metabolism; D003587:Cytomegalovirus / Q000235:genetics; D005260:Female; D005315:Fetal Diseases / Q000378:metabolism / Q000628:therapy*; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000008:administration & dosage*; D005865:Gestational Age; D006023:Glycoproteins / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000097:blood / Q000235:genetics / Q000378:metabolism; D015497:HIV-1 / Q000235:genetics*; D007274:Injections, Intraperitoneal; D009052:Leukemia Virus, Murine / Q000235:genetics; D008099:Liver / Q000378:metabolism; D008253:Macaca mulatta; D008856:Microscopy, Fluorescence; D011401:Promoter Regions, Genetic; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014161:Transduction, Genetic / Q000379:methods*; D019076:Transgenes; D014721:Vesicular stomatitis Indiana virus / Q000235:genetics
D006023:Glycoproteins; D049452:Green Fluorescent Proteins