Lentiviral vectors containing the human immunodeficiency virus type-1 central polypurine tract can efficiently transduce nondividing hepatocytes and antigen-presenting cells in vivo.
High-titer self-inactivating human immunodeficiency virus type-1 (HIV-1)-based vectors expressing the green fluorescent protein reporter gene that contained the central polypurine and termination tract and the woodchuck hepatitis virus posttranscriptional regulatory element were constructed. Transduction efficiency and biodistribution were determined, following systemic administration of these improved lentiviral vectors. In adult severe combined immunodeficiency (SCID) mice, efficient stable gene transfer was achieved in the liver (8.0% +/- 6.0%) and spleen (24% +/- 3%). Most transduced hepatocytes and nonhepatocytes were nondividing, thereby obviating the need to induce liver cell proliferation. In vivo gene transfer with this improved lentiviral vector was relatively safe since liver enzyme concentration in the plasma was only moderately and transiently elevated. In addition, nondividing major histocompatibility complex class II-positive splenic antigen-presenting cells (APCs) were efficiently transduced in SCID and normal mice. Furthermore, B cells were efficiently transduced, whereas T cells were refractory to lentiviral transduction in vivo. However, in neonatal recipients, lentiviral transduction was more widespread and included not only hepatocytes and splenic APCs but also cardiomyocytes. The present study suggests potential uses of improved lentiviral vectors for gene therapy of genetic blood disorders resulting from serum protein deficiencies, such as hemophilia, and hepatic disease. However, the use of liver-specific promoters may be warranted to circumvent inadvertent transgene expression in APCs. In addition, these improved lentiviral vectors could potentially be useful for genetic vaccination and treatment of perinatal cardiac disorders.
Center for Transgene Technology and Gene Therapy, Flanders Interuniversity Institute for Biotechnology-University of Leuven, Belgium. thierry.vandendriessche@med.kuleuven.ac.be
Thierry
T
VandenDriessche
Lieven
L
Thorrez
Luigi
L
Naldini
Antonia
A
Follenzi
Lieve
L
Moons
Zwi
Z
Berneman
Desire
D
Collen
Marinee K L
MK
Chuah
D000818:Animals; D000938:Antigen-Presenting Cells / Q000378:metabolism*; D001682:Biological Availability; D002455:Cell Division; D004247:DNA / Q000737:chemistry; D005809:Genes, Regulator; D005822:Genetic Vectors / Q000008:administration & dosage / Q000235:genetics* / Q000493:pharmacokinetics*; D015497:HIV-1 / Q000235:genetics*; D017919:Hepatitis B Virus, Woodchuck / Q000235:genetics; D022781:Hepatocytes / Q000378:metabolism*; D006801:Humans; D016086:Lentivirus / Q000737:chemistry / Q000235:genetics; D008099:Liver / Q000166:cytology / Q000378:metabolism; D051379:Mice; D016513:Mice, SCID; D011687:Purines / Q000737:chemistry*; D012191:Retroviridae Proteins / Q000032:analysis; D013154:Spleen / Q000166:cytology / Q000378:metabolism; D014018:Tissue Distribution; D014161:Transduction, Genetic / Q000379:methods*
D011687:Purines; D012191:Retroviridae Proteins; D004247:DNA