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

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. 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 micro g/mL protamine sulfate, the cells were cultured in a transforming media prior to differentiating into DCs by GM-CSF, TNF-alpha 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. Titration of viral vectors indicated a transduction efficiency of 1 x10[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 forCD86+ HLA-DRand CD1alpha+CD14, respectively. 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

Study on the frequent expression of rodgers and chido, red blood cell antigens, in iranian healthy population

Rodgers [Rg] and Chido [Ch] are blood-group antigens and they determine the fourthcomponent of human complement C4. Rodgers and Chido are associated with two C4 isotypes [C4A and C4B]. In addition to genotype determination, study on expression of Rg and Chido could be useful in disease studies. DNA was extracted from the whole blood of 60 normal individuals. Then, PCR amplification of C4d gene fragment was followed by restriction digestion. This study demonstrated that the frequency of Ch and Rg in Iranian healthy population was 98.3 and 93.4 percent, respectively. Additionally, 6.6 percent of the studied population showed Chido-positive, Rodger-negative and 1.7 percent showed Rodger-positive, Chido-negative genotype. It may be concluded that upon receiving blood transfusion, 6.6 and 1.7 percent of individuals could produce anti-Rg and anti-Ch antibodies, respectively