The Experimental Study of Culturation of Dendritic Cells from Peripheral Blood in Patients with Chronic Hepatitis B

Objective To culture dendritic cells(DCs)from p er ipheral blood monocytes(PBMC)in patients with chronic hepatitis B(CHB)in vitro ,and offer the experimental background to explore the action in CHB immune toler ance and clinical application of DCs.Methods In 10 patients with CH B,PBMC was isolated from 15 ml peripheral blood of each patient.The DCs were cul tured with existences of Granulocyte/Macrophage Colony-stimulating Factor(GM- CSF)and Interleukin 4(IL0-4).Electron microscope was employed to analyse th e morphology of DCs.The expression of phenotypic molecules of DCs was detected w ith flow cytometry.Results ①A combination of GM-CSF and IL-4 pro vided about (1.2～4.7)×106 DCs from 15ml blood in patients with CHB after cul t ured for 9 days,whose morphology was tested by electron microscope.②The expres sion of phenotypic molecules of DCs was very weak,with CD83(8.02±3.99)%、CD8 0(8.77±2.06)% and MHC-DR(14.05±2.66)%.Conclusion DCs culture d from PBMC in the patients with CHB under the conditions of GM-CSF and IL-4 p resent typical dendritic morphology but are in the immature state for expressing phenotypic molecules.