Biological effects of RelB gene silencing on the dendritic cells derived from bone marrow / 解放军医学杂志

ABSTRACT

Objective To study the biological effects of nucleic transcript factor NF-?B/RelB gene silencing on bone marrow derived dendritic cells(BMDCs).Methods For constructing a new type of DCs,RNAi RelB DC,mouse BMDCs were cultured through IL-4 and GM-CSF cytokine inducing system and were purified with CD11c positive immunomicrobeads by flow cytometer,and RelB gene was silenced with PCR expression cassettes.The RelB RNA interference effect in DCs was proved at both mRNA and protein levels.The biological characteristics of RNAi RelB DCs were investigated by means of cell morphology,cell surface markers and mixed lymphocyte reaction ability.The cell microstructure was observed using scanning electron microscopy and transmission electron microscopy.The cell surface markers,such as MHC-Ⅱ,CD86 and CD40,were detected by flow cytometer.The mixed lymphocyte reaction ability,as well as the ability of DCs in stimulating allogeneic T cells proliferation,was detected by 3H-TdR incorporation method.Results RNAi RelB DCs seemed to have fewer and shorter spikes and many folds under scanning electron microscopy.Many phagocytic vacuoles were observed in the cytoplasm under transmission electron microscopy.DCs surface molecules,MHC-Ⅱ,CD86 and CD40,were expressed at a relatively lower level compared with the control DCs.This RNAi RelB DCs also showed weak allogeneic T cells stimulating ability in vitro in MLR.Conclusion The dendritic cells derived from bone marrow are characterized the biological features and immunological function of immature DC with silencing of RelB gene.This kind of RNAi RelB DCs may be used to induce immunotolerance as a novel tolerogenic DCs.