摘要
目的探讨Toll样受体3(TLR3)介导的天然免疫对Bewo细胞中乙型肝炎病毒(HBV)复制的影响。方法首先用TLR3配体polyI:C处理Bewo细胞,观察细胞TLR3 mRNA表达的动力学。然后将2μg 1.3倍HBV全基因重组质粒pcDNA3.1(+)-HBV1.3转染Bewo细胞,12h后,以polyI:C处理3d。最后,用抗TLR3处理Bewo细胞1h后,加入25μg/ml polyI:C刺激。采用荧光定量RT-PCR、微粒子酶免疫分析法(MEIA)和荧光定量PCR法分别检测细胞TLR3mRNA表达、HBsAg、HBeAg及HBV DNA水平。结果 polyI:C可显著诱导Bewo细胞TLR3 mRNA表达(P<0.05),呈时间和剂量依赖性;与对照组比较,polyI:C可显著抑制HBV复制(P<0.001),抗TLR3可显著逆转polyI:C对HBV复制的抑制作用(P<0.01)。结论 TLR3介导的天然免疫能一定程度抑制Bewo细胞中HBV复制,为防治HBV宫内感染提供了新的途径。
Objective：To explore the effect of TLR3-mediated innate immune on hepatitis B virus（HBV） replication in Bewo cells.Methods： Firstly,Bewo cells were exposed to TLR3 ligand polyI：C to observe the kinetics of TLR3 mRNA expression in Bewo cells.Secondly,2μg 1.3-fold HBV recombinant vector pcDNA3.1（＋）-HBV1.3 were transfected into Bewo cells,after 12h,the cells were treated with polyI：C for 3d.Eventually,after Bewo cells were stimulated with anti-TLR3 antibody for 1h,then treated with 25μg/ml polyI：C.To assay TLR3 mRNA expression in Bewo cells by real-time fluorescence quantitative RT-PCR（qRT-PCR）.HBsAg,HBeAg and HBV DNA level was detected by Microparticle Enzyme Immunoassay（MEIA） and fluorescence quantitative PCR,respectively.Results： PolyI：C could significantly induce TLR3 mRNA expression in Bewo cells（P0.05）,in time-and dose-dependent manners.Compared with control,polyI：C could significantly suppressed HBV replication（P0.01）.The suppressive effect of polyI：C on HBV replication was reversed by neutralizing antibodies against human TLR3（P0.01）.Conclusions：TLR3-mediated innate immune could suppressed hepatitis B virus replication in Bewo cells to some degree,and this will provide a new way for prevention and cure intrauterine HBV infection.
出处
《中国优生与遗传杂志》
2011年第10期16-18,111,共4页
Chinese Journal of Birth Health & Heredity
