Gibbon ape leukemia virus transduction of peripheral blood CD34 + -derived dendritic cells

BACKGROUNDDendritic 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 asessed the efficacy of GALV transduction of CD34 + derived DCs as a possible approach to creating genetically modified DCs for immunotherapy.MATERIALS AND METHODSPeripheral 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 pg/mL protamine sulfate, the cells were cultured in a transforming media prior to differentiating into DCs by GM-CSF, TNF-α 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.RESULTSTitration of viral vectors indicated a transduction efficiency of 1 × 105 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 CD1 α + CD14, respectivelyCONCLUSIONThis 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.