The Function Of Dendritic Cells Derived From Peripheral Blood Mononuclear Cells In Patients With ITP

Objective To investigate the proliferation and functions of dendritic cells (DC) derived from PBMC in patients of idiopathic thrombocytopenic purpura(ITP), a comparison was made in the differences in phenotypes and the stiumlating capacities of allogeneic lymphocytes between patients with ITP and those of normal controls.Methods The monouclear cells were prepared from peripheral blood mononuclear cell(PBMC) with lymphoprep density gradient centrifugation method, then induced with the recombinant cytokines rh GM-CSF(100 ng/ ml), rh IL -4(500 U/ ml), and LPS (50U/ml) for six day. we observed the DCs growth under the microscope every day.And we examine the cellular construction under the transmission electron microscope and scanning electronmicroscopy and determined DCs phenotypes by flow cytometry at 6d. The capacity of DCs to activate allogeneic lymphocytes immune responses was tested by MTT.The concentrations of IL-12 in the supernatants of DC cultures was determined by ELISA at 6d.Thirty eight patients with ITP were included in this study, twenty three healthy subjects being used as a control group.Results The dendritic cells cultured in vitro turned into suspensive growth from adhesive situation at 5d. Then the number of DCs increased continuously and the cells showed the irregular morphologic appearance of DCs with veiled edges on the 6th day.It was showed by flow cytometry that the mature DCs expressed the relatively specific marker such as CD80 and CD80. The expression levels of CD80 and CD86 on DC surface from patients were significantly higher than those of normal controls. Similarly, the capacity of DC to stimulate the allogeneic T lymphocytes was apparently enhanced in ITP patients. After 6 days culturing, the concentrations of IL-12 determined by ELISA in ITP patients was much higher than those of normal controls.Conclusion â‘ The dendritic cells can be induced from human peripheral blood mononuclear cell and induced into mature through the cultivation with rh GM-CSF, rh IL -4 and LPS in vitro. â‘¡Compared to normal controls, the state of dendritic cells in ITP patients is appreciably matured. â‘¢The differentiation of monocytes into dendritic cells is upregulated in ITP, and these cells have more potent antigen presenting capacities than those of normal controls.