The Effection Of Hepatitis B Virus On Monocyte-Derived Dendritic Cell Phenotype And Autophagy

Objective:To investigate the effect of hepatitis B virus (HBV) infection on autophage of monocyte-derived dendritic cells (DCs), and dendritic cells phenotype changes with the change of autophagy.Materials and methods:Fresh peripheral blood was obtained from healthy donors and the patients with chronic HBV infection, and monocytes were purified by positive selection using anti-CD 14-conjugated magnetic microbeads. Immature DCs were obtained after 6-day cultured, then we used pure 1640 culture (control group) and lipopolysaccharide (LPS) (LPS group), HepG2.2.15 supernatant (HBV group), HepG2 supernatant (HepG2 group) to prompt imDC to maturity. Following stimulation of autophagy, we used flow cytometry to detect the change of DC phenotype, used western blot, immunofluorescence technique to detect autophagy related protein (LC3,Atg-5).Results:After 10-day cultured, the diameters of DCs were bigger than the precursors-mononuclear cells, and the intracellular particles of 10-day cultured DCs were significantly more than that of the precursors. Compared with control group, DCs derived from healthy donors had high expression of HLA-DR, CD80, CD83 and CD86, however, the level of CD83 was lower than LPS group and HLA-DR, CD80, CD86 were higher than LPS group. For derived from the patients with chronic HBV infection, compared with control group, DCs expressed higher level of HLA-DR, CD80, CD86 and lower level of CD83(P<0.05). According to the fluorescent confocal microscope and western blot, after stimulated by HBV, DCs derived from healthy donors and chronic HBV infection patients both had higher expression of LC3 and Atg-5 compared to control group (p<0.05).Conclusions:After stimulation, there was the same trend for autophage of DC from healthy donors and chronic HBV infection patients. Persistent HBV infection has no obvious influence on morphology and particle of DCs derived from mononuclear cells. After infected with HBV, the autophagy of DCs was enhanced and the HLA-DR, CD80, CD86 expression levels were increased, the expression level of CD83 was decreased.