Different HBsAg Concentrations Loading On Dendritic Cells From Peripheral Blood Monocytes In Patients With Chronic Hepatitis B Effects The Cell Phenotypic Molecules Expression

ObjectivesTo culture dendritic cells(DCs)from peripheral blood monocytes(PBMC) in patients with chronic hepatitis B(CHB)in vitro,and observe the changes of the phenotypic molecules on DCs loaded by different HBsAg concentrations to offer the theoretical background of CHB treatment with DCs. Methods 40 patients with CHB were divided randomly into 4 groups with 10 per group. Peripheral blood monocytes from these patients were isolated,and DCs were cultured under the conditions of Granulocyte/Macrophage Colony-stimulating Factor(GM-CSF)and Interleukin 4(IL-4)existences. DCs of the experimental groups were loaded by three different HBsAg concentrations , 2.5mg/L,5mg/L and 10mg/L, for 24 hours while the control group not. Electron microscope was employed to analyse the morphology of DCs. The expression of phenotypic molecules of DCs was detected with flow cytometry.Results 1 A combination of GM-CSF and IL-4 provided about 1.2-4.7×106 DCs from 15ml blood in patients with CHB after cultured 9 days,whoes morphology was tested by electron microscope. 2 The expression of phenotypic molecules of DCs in control group was very low,with CD83(8.02±3.99)%,CD80(8.77±2.06)% and MHC-DR(14.05±2.66)%. 3 After loaded by different HBsAg concentrations, the upregulation of phenotypic molecules of DCs was found,with CD83(18.35±2.93)%,CD80(42.63±7.15)% and MHC-DR(47.49±6.59)% in HBsAg 2.5mg/L group； CD83(17.88±3.12)%,CD80(45.24±10.93)% and MHC-DR(47.07±8.52)% in HBsAg 5mg/L group and CD83(16.74±2.86)%,CD80(44.59±6.99)% and MHC-DR(48.59±7.42)% in HBsAg 10mg/L group respectively. Compared with control group,the experimental groups are all different remarkably(p<0.01),but among them,there were no differences(p>0.05)Conclusions 1 DCs cultured from PBMC in the patients of CHB under the conditions of GM-CSF and IL-4 present on the typical dendritic morphology but are in the immature state for expressing phenotypic molecules very low.2 After loaded by different HBsAg concentrations,the immature DCs can differentiate to mature DCs with the upregulation of the expression of phenotypic molecules,which may potentialize the function of antigen presenting and activate immune reponse.3 Immature DCs take up antigens potently and become mature DC.4 HBsAg loading on DC,which may improve the function of antigen presenting,is a promising and effective way to reverse CHB specific immune tolerance and explore a novel immune therapy for CHB.