The Immunologic Mechanism Of Dendritic Cells Vaccine In Hepatitis B Infected Person

chapter 1HBV core and nuclear antigen can enhance the differation and function of human dendritic cellsObjectiveMonocyte-derived dendritic cells (MoDCs) are a promising cellular adjuvant for effector immune responses against tumors and chronic viral infections, including HBV. Several factors can induce DC maturation and activation including microorganisms, bacterial and viral products and cytokines. HBV core and nuclear antigen(HBcAg and HBeAg) are the most important antigens of HBV. We thus endeavored to analyze the ability of these two antigens to enhance the maturition of human DCs and the secretion of IFN-γand IL-12p70 by activated DCs, and to evaluate the influence of HBcAg and HBeAg on Human DCs in the anti-HBV pathoimmunological mechanisms.MethodsThe DC populations were generated from peripheral blood mononuclear cells(PBMCs) derived from healthy donors and then pulsed with HBcAg and HBeAg respectively.Cell surface molecules, including CD83, CD86 and HLA-ABC, were analyzed with flow cytometry. Concentrations of IFN-γ,IL-12p70 in the supernatant were assayed by ELISA methods.Results Upregulation of maturation-associated phenotypic markers ( human leucocyte antigen HLA-ABC, CD83, CD86 ) on Maturity Dendritic Cell(mDC) after 72 h HBcAg/HBeAg pulsing was observed. The lymphocyte proliferation, the cytotoxic activity,as well as the IFN-γand IL-12p70 secretion, were observed more sronger in the HBcAg and HBeAg pulsed DCs than in the mDCs non-stimulated. And, HBcAg is more powerful than HBeAg.ConclusionHBcAg and HBeAg can enhance the maturation and secretion of human dendritic cells,and can improve lympholeukocyte multiplication and cytotoxic T lymphocyte activity.chapter 2The influence of IL-10 and TNF-αon the regulation of human dendritic cells in chronic hepatitis B patientsObjectiveTo investigate the influence of IL-10 and TNF-αon the regulation of human dendritic cells in chronic hepatitis B(CHB) patients.MethodsMonocytes were isolated from fresh peripheral blood of CHB patients by Ficoll-Hypaque density gradient centrifugation and cultured with plastic-adherence method. DCs were induced and proliferated from the monocytes with granulocyte-macrophage clony stimulating factor(GM-CSF) and interleukin-4(IL-4)for Seven days, and then cocultured with TNF-αand IL-10 respectively. Elements reflected the maturity of DC were detected. The lymphocyte proliferation, as well as the IL-12p70 secretion were observed.ResultsUp-regulation of maturation-associated phenotypic markers ( human leukocyte antigen: HLA-A, B, C, HLA-DR, CD86)on Maturity Dendritic Cells(mDC) after TNF-αpulsing was observed.The lymphocyte proliferation, as well as the IL-12p70 ,IL-21 secretion were observed more stronger in the TNF-αpulsed DCs than in the DCs non-stimulated(P<0.01).The secretion and expression of human leukocyte antigens, as well as the lymphocyte proliferation by DCs were inhibited while treated with IL-10.ConclusionThe number, maturation and function of DCs from CHB patients were all lower than from healthy donors. But the DCs from CHB patients can be regulated effectively in vitro. TNF-αcan promote DC ripe for mDC, while IL-10 inhibit DCs'maturation.