Objective To detect the expression of type Ⅰ interferon in monocyte-derived dendritic cells(MoDCs)after Toll like receptor(TLR)3 triggered in patients with chronic hepatitis B(CHB),and to evaluate immune responses of CHB patients and its roles in the mechanisms of persistent infection of hepatitis B virus(HBV)and chronicity of hepatitis.Methods Peripheral blood mononuclear cells(PBMCs)were isolated and purified using magnetic beads(plasma was saved simultaneously)from 26 CHB patients and 18 healthy volunteers(HV).Dendritic cells(DCs)were induced and proliferated in a culture medium with recombinant human granulocyte macrophage colony stimulating factor(rhGM-CSF)and recombinant human interleukin(rhIL-4).EX3s were stimulated with Poly Ⅰ:C and the supernatants were collected at 0 h and 24 h after stimulation.Type Ⅰ interferon(IFN-α and IFN-β)in plasma and supernatants were examined by enzyme linked immunosorbent assay (ELISA).Results The levels of type Ⅰ interferon in plasma were not significantly different in groups of HV and CH B.IFN-α and IFN-β expressions in supernatants before Poly Ⅰ:C stimulation were(80.00±16.15)ng/L,(36.39±13.90)ng/L in CHB group and(76.76±15.90)ng/L,(37.14±13.68)ng/L in HV group,respectively.And there were no statistical differences between two groups(t=1.651,t=0.178;both P>0.05).IFN-α expressions in supernatants at 24 h after stimulation in two groups were both higher than those before stimulation(at 0 h),but there were no statistical differences(t=1.534,t=1.243;both P>0.05).IFN-β expressions in supernatants at 24 h after stimulation in HV group was(54.57±16.80)ng/L,which was significantly higher than that at 0 h(37.14±13.68)ng/L(t=4.061,P<0.05).However,there was no significant difference at 24 h than tht at 0 h in CHB group(t=1.796,P>0.05).At 24 h after stimulation.IFN-β level was(54.57±16.80)ng/L in HV group,which was significantly higher than that[(41.64±12.57)ng/L]in CHB group(t=2.921,P<0.05).Conclusions Functions of MoDCs from CHB patients are impaired and MoDCs could not express type Ⅰ interferon normally.Expression of type Ⅰ interferon after TLR3 triggered in CHB patients is mainly IFN-β. 相似文献
Objective To detect the secretions of type Ⅰ interferon and the expressions of phospho-IRF3 in murine liver dendritic cells intervened by HBV.Methods The murine liver dendritic cells were isolated via anti-CD11c microbeads and were incubated in the presence of GM-CSF and IL-4 to induce the DC generation and proliferation in 24-well cell culture plates.HBV virions were isolated via ultracentrifugation and were detected by quantitative Realtime-PCR.The DCs were divided into two groups:one group was cultured with HBV virions for 24 hours,the other group was cultured without HBV as control group.The cells were harvested at Oh,1h,2h,6h and 24h after being stimulated with poly I:C and the expressions of pIRF3 and the concentration of IFN β in supematants were detected with western blot and ELISA respectively.Results The IFN β concentrations at 0h,6h and 24h in the supernatants of the RBV group and the control group were(12.38±3.71)pg/ml,(88.67±9.01)pg/ml and(69.89±5.80)pg/ml vs(10.83±4.11)pg/ml,(137.68 ± 12.28) pg/ml and (72.25 ± 8.61) pg/ml,respectively. No statistical differences found at 0 h (t =0.8398,P > 0.05) and 24 h (t = 0.6820,P > 0.05) between the two groups except that at 6 h (t = 9.653,P <0.01). The expressions of phospho-IRF3 in HBV group were lower than that in control group. Conclusions The type Ⅰ interferon secretion and the phospho-IRF3 expression were decreased in murine liver dendritic cells when intervened by HBV. 相似文献