摘要目的 研究HBV干预后小鼠肝脏树突状细胞P-干扰素调节因子3及其下游Ⅰ型干扰素表达的变化.方法用免疫磁珠分选法分离肝脏树突状细胞,并用粒细胞-巨噬细胞集落刺激因子和白细胞介素4在24孔板中诱导培养.用超离心技术获得HBV,并用实时荧光定量PCR检测.第5天将培养的树突状细胞分为2组:HBV组与HBV共同培养,对照组加入等量的细胞培养液.24 h后,两组均加入聚肌胞刺激,收集0、1、2、6、24 h的细胞和上清液,用Western blot检测P-干扰素调节因子3的表达情况,用酶联免疫吸附法检测上清液中干扰素β的浓度.组间比较用t检验.结果 0h时HBV组培养液上清液干扰素β浓度[(12.38 ±3.71)pg/ml]与对照组[(10.83±4.11)pg/ml]比较,差异无统计学意义(P>0.05).聚肌胞刺激后6 h,HBV组培养液上清液干扰素β浓度[(88.67±9.01)pg/ml]与对照组[(137.68±12.28)pg/ml]比较,t=9.653,P<0.01,差异有统计学意义.聚肌胞刺激后24h,HBV组培养液上清液干扰素β浓度[(69.89±5.80)pg/ml]与对照组[(72.25±8.61)pg/ml]比较,差异无统计学意义(P>0.05).聚肌胞刺激小鼠肝脏树突状细胞后P-干扰素调节因子3表达逐渐增强.经HBV干预在感染时病毒与细胞数量的比值为25,24 h后再用聚肌胞刺激2 h,HBV组P-干扰素调节因子3表达较对照组减弱.结论 HBV干预后小鼠肝脏树突状细胞P-干扰素调节因子3及其下游Ⅰ型干扰素表达均下调.

abstractsObjective 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.