Primary study on relationship between dendritic cells and differentiation of CD_4~+Th cell sin patients with chronic hepatitis B

ObjectiveTo survey the relationship between dendritic cells (DCs) and differentiation of CD4+ cell subgroups in patients with chronic hepatitis B (CHB). MethodsThe peripheral blood mononuclear cells (PBMCs) were isolated and then cultured with rhIL-4 (50 ng/mL), rhGM-CSF (10 ng/mL) and rhTNF-α (100 μ/mL) to derive DCs. The phenotypes of DCs including CD1a, CD83, CD80, CD86 and HLA-DR were detected by flow cytometer (FCM). The CD4+ T cell subpopulation in peripheral blood was separated by immunomagnetic beads, and then Th cells were stained with fluorescent staining after PMA+Ionomycin stimulation, and the differentiation of Th1/Th2 was observed by flow cytometry detection of intracellular IFN-γ and IL-4. The contents of IL-6, IL12, IFN-γ and IL-4 in culture supernatants of DCs or Th cells were respectively detected with ELISA. Results The expression of CD1a, CD80, CD83, CD86 and HLA-DR in the DCs of the CHB patients was markedly lower than that of health adults. On the 7th culture day, the concentration of IL-12 in DC supernatants of CHB patients was lower than that of health adults, but the concentration of IL-6 was higher. In comparison with health adults, the percentages of Th1 cells in CD4+ T cells in peripheral blood of CHB patients and the contents of IFN-γ in Th1 cell supernatants of CHB patients were both lower. When the CHB patients′ DCs were co-cultured with Th cells from allogenic health adults, their capability of making Th1 cells to generate IFN-γ was remarkably weaker than that of health adults′ DCs. ConclusionsIn patients with CHB, insufficiency of DCs′ function possibly causes deficiency of Th1 cell differentiation in peripheral blood.