Induction Culture Of Tolerogenic Dendritic Cells From Human Peripheral Blood And Their DC-STMP Expression

［Objectives］To induce and culture the tolerogenic dendritic cells(TolDCs)from human peripheral blood, obverse the dendritic cells-specific transmembraneprotein (DC-STAMP) mRNA expression levelduring the TolDCs inductionculture.［Methods］(1)CD14+monocytes were purified from Peripheral blood mononuclearcells (PBMNC) in immunomagnetic bead sorting system and cultured in themedium containing Granulocyte-macrophage colony-stimulating factor (GM-CSF)，Interleukin-4(IL-4)，Dexamethasone sodium phosphate (Dex)， Lipopolysaccharide(LPS),inducing them into TolDCs.(2) Cultured cells morphology andCD83,CD86and CD14flow cytometry were followed. Mixed lymphocyte reaction(MLR) was performed to measure their capacity inducing T lymphocyteproliferation.(3) Real time RT-PCR was used to track DC-STAMP mRNAexpression level during TolDCs induction culture.［Results］(1) CD14+monocytes induction culture with Granulocyte-macrophagecolony-stimulating factor (GM-CSF)，Interleukin-4(IL-4)，Lipopolysaccharide (LPS)exhibited typical morphologic of mature dendritic cells, the cells wasshort of the typical morphologic when treated with Dex.(2) CD14+monocytesinduction culture with GM-CSF， IL-4led to higher expression of CD83,CD86and lower expression of CD14. When they were treated with Dex theexpression of CD83,CD86was downregulation and the expression of CD14wasupregulation, their capacity to stimulate allogeneic T cells wassignificantly decreased,exhibited typical functional characteristics andimmunological phenotype of TolDCs.(3) The expression level of DC-STAMPmRNA decreased during dendritic cells culture. However, the expressionincreased when they were treated with Dex and induced to from TolDCs.［Conclusions］Culture of CD14+monocytes from human peripheral blood in the medium containing GM-CSF,IL-4,Dex and LPS can successful induce TolDCsformation.DC-STAMP may be associated with the tolerance function of DC.