Improved short-term engraftment of lentivirally versus gammaretrovirally transduced allogeneic canine repopulating cells.
Gammaretroviral vectors require cell division for efficient transduction. Thus, extended cell culture times are necessary for efficient transduction with gammaretroviral vectors, which in turn can lead to stem cell loss and impaired engraftment. Lentiviral vectors transduce nondividing cells and are therefore able to transduce stem cells in short transduction protocols. Here, we compared the short-term engraftment of lentivirally and gammaretrovirally transduced canine allogeneic DLA-matched littermate cells. A reduced conditioning regimen of 400 cGy total body irradiation was used in preparation for clinical studies. Two dogs received a graft of gammaretrovirally transduced CD34-selected cells. CD34(+) cells were prestimulated for 30 h and then exposed twice to concentrated RD114 pseudotype vector. Three dogs received lentivirally transduced CD34-selected cells. Cells were transduced overnight with concentrated VSV-G pseudotype lentiviral vector. The animals in the lentiviral group showed a significantly faster granulocyte recovery. VNTR analysis 40-50 days after transplantation revealed higher donor chimerism for the lentiviral group compared to the retroviral group. These data suggest that short lentiviral transduction protocols may be superior to extended gammaretroviral transduction protocols with respect to engraftment potential of transduced CD34(+) hematopoietic repopulating cells.
Neff
Tobias
T
Clinical Research Division, Fred Hutchinson Cancer Research Center, Seattle, WA, USA.
Gerull
Sabine
S
Peterson
Laura J
LJ
Kiem
Hans-Peter
HP
D000818:Animals; D018952:Antigens, CD34; D002478:Cells, Cultured; D004285:Dogs; D017988:Gammaretrovirus / Q000235:genetics*; D006085:Graft Survival; D006098:Granulocytes / Q000502:physiology; D018380:Hematopoietic Stem Cell Transplantation / Q000379:methods* / Q000592:standards; D007700:Kinetics; D016086:Lentivirus / Q000235:genetics*; D014161:Transduction, Genetic / Q000379:methods*; D018183:Transplantation Chimera; D019172:Transplantation Conditioning / Q000379:methods; D014184:Transplantation, Homologous
D018952:Antigens, CD34