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Antibody to E1 peptide of hepatitis C virus genotype 4 inhibits virus binding and entry to HepG2 cells in vitro

机译:丙型肝炎病毒基因型4的E1肽抗体在体外抑制病毒结合和进入HepG2细胞

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摘要

AIM: To analyze the neutralizing activity of antibodies against E1 region of hepatitis C virus (HCV). Specific polyclonal antibody was raised via immunization of New Zealand rabbits with a synthetic peptide that had been derived from the E1 region of HCV and was shown to be highly conserved among HCV published genotypes.METHODS: Hyper-immune HCV E1 antibodies were incubated over night at 4 °C with serum samples positive for HCV RNA, with viral loads ranging from 615 to 3.2 million IU/ mL. Treated sera were incubated with HepG2 cells for 90 min. Blocking of viral binding and entry into cells by anti E1 antibody were tested by means of RT-PCR and flow cytometry.RESULTS: Direct immunostaining using FITC conjugated E1 antibody followed by Flow cytometric analysis showed reduced mean fluorescence intensity in samples pre-incubated with E1 antibody compared with untreated samples. Furthermore, 13 out of 18 positive sera (72%) showed complete inhibition of infectivity as detected by RT-PCR.CONCLUSION: In house produced E1 antibody, blocks binding and entry of HCV virion infection to target cells suggesting the involvement of this epitope in virus binding and entry. Isolation of these antibodies that block virus attachment to human cells are useful as therapeutic reagents.
机译:目的:分析抗丙型肝炎病毒(HCV)E1区抗体的中和活性。通过用源自HCV E1区的合成肽免疫新西兰兔来产生特异性多克隆抗体,该合成肽在HCV已发表的基因型中显示出高度保守的方法。 4°C,HCV RNA阳性血清样品,病毒载量范围为615至320万IU / mL。将处理过的血清与HepG2细胞孵育90分钟。通过RT-PCR和流式细胞术检测了抗E1抗体对病毒结合的阻断和进入细胞的结果。结果:使用FITC缀合的E1抗体直接免疫染色,然后用流式细胞术分析显示,用E1预孵育的样品平均荧光强度降低抗体与未处理样品相比。此外,通过RT-PCR检测到,在18份阳性血清中有13份(72%)显示出完全抑制了感染性。结论:在内部产生的E1抗体中,阻断了HCV病毒体感染与靶细胞的结合和进入,表明该表位参与了病毒绑定和进入。阻断病毒附着于人细胞的这些抗体的分离可用作治疗剂。

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