Gibbon ape leukemia virus transduction of peripheral blood CD34+-derived dendritic cells.
BACKGROUND
Dendritic cells (DCs) play a critical role in the immune response and are a candidate for immunotherapy in cancer. Since gibbon ape leukemia virus (GALV) transduction of CD34+ cells is reasonably efficacious, we assessed the efficacy of GALV transduction of CD34+ derived DCs as a possible approach to creating genetically modified DCs for immunotherapy.


METHODS
Peripheral blood CD34+ cells were transduced with retroviruses obtained from the PG13/LN C8 cell line, with the neomycin gene as a marker gene. After prestimulation of hematopoietic cells for 24 hours with 10 ng/mL interleukin (IL)-3, 10 ng/mL IL-6, 100 ng/mL stem cell factor, 100 ng/mL granulocyte-macrophage colony stimulating factor and 8 Amicrog/mL protamine sulfate, the cells were cultured in a transforming media prior to differentiating into DCs by GM-CSF, TNF-a and IL-4. Immunophenotyping analyses for confirmation of the generated DCs, colony formation assay and PCR were done for the expression of neomycin gene in the transduced cells.


RESULTS
Titration of viral vectors indicated a transduction efficiency of 1×10(5) CFU/mL. Transduction efficiency for the CD34+ cells transformed to DCs was 45% and 38% before and after DC differentiation, respectively. Additionally, a mean (SEM) of 26.9% (11.4%) and 41.4% (11.8%) of the genetically modified DCs were positive for CD86+ HLA-DR and CD1a+CD14, respectively


CONCLUSIONS
This study showed that the majority of transduced CD34+ cells were successfully differentiated into cells identical to DCs according to morphology and immunophenotyping features, which could be a potential application in immunotherapy.
Faculty of Paramedical Sciences, Tehran University of Medical Sciences, Tehran, Iran.
Leili
L
Moezzi
Kamran
K
Alimoghaddam
Alireza J
AJ
Saleh
Somayeh
S
Shahrokhi
Seyes Hamidolah
SH
Ghaffari
Nadia
N
Bagheri
Bahram
B
Chardouli
Ardeshir
A
Ghavamzadeh
D018952:Antigens, CD34; D001773:Blood Cells; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation; D003713:Dendritic Cells / Q000166:cytology / Q000378:metabolism*; D006412:Hematopoietic Stem Cells / Q000166:cytology; D006801:Humans; D007167:Immunotherapy / Q000379:methods*; D017989:Leukemia Virus, Gibbon Ape / Q000235:genetics*; D014161:Transduction, Genetic / Q000379:methods*
D018952:Antigens, CD34