Lentivirus Mediated Blimp1-shRNA Gene Therapy Regulates The Differentiation Of Bone Marrow-derived Dendritic Procursors(Pre-DCs)

Objective: Lenti virus mediated Blimp1shRNA gene therapy regulates Blimp1geneexpression on dendritic cells (DCs) differentiated from bone marrow cells(BMCs)Method: BMSCs from Balb/c mice were induced differentiation to DCs in a8day cellculture system with GM CSF/IL4incubation and LPS overnight stimulation at day7. TheDCs were identified by CD11c and quantitied CD86/MHC II for maturation using flowcytometry at day8. At day2, lenti Blimp1gene therapy was given in the culture system,while no therapy and lenti virus GFP therapy were given as empty control and lenti control,respectively. From day4, transfection efficiency was measured by fluorescent.72h and96hafter transfection, Blimp1mRNA was analyzed by real time RT PCR and AXL protein wasdetected by Western blot.Result: GFP fluorescent showed lenti virus transfection efficiency of3day is30.4±4.8%,there are obvious differences when compare two transfected groups with the empty controlgroup in cell culture curves. The Blimp1mRNA and protein in the lenti Blimp1group wereexpressed1%and74%compared with the empty control group. At day8, in theempty control group, CD11c positive cells and CD86/MHC II positive cells were69.24±4.98%,51.08±4.93%and56.28±7.30%. While68.56±5.86%,49.52±4.28%and69.38±4.52%in the lenti control group, and72.76±5.52%,50.20±6.01%and46.48±5.70%in the lenti Blimp1group.Conclusion: Antisense Blimp1gene therapy was down regulated Blimp1expression onDCs to moderate the following allograft immune reaction. lentivirus mediatedBlimp1shRNA gene therapy depress DC maturation during the differentiation progressfrom BMSCs. The ex vivo and in vivo experiment for the allograft immune reaction needfurther investigation.