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An influenza A virus hemagglutinin (HA) epitope inserted in and expressed from several loci of the infectious bursal disease virus genome induces HA-specific antibodies

机译:从传染性法氏囊病病毒基因组的几个基因座插入并表达的甲型流感病毒血凝素(HA)表位诱导HA特异性抗体

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The N-terminus of infectious bursal disease virus (IBDV) VP5 has been shown to be capable of tolerating the insertion of small epitopes. The objective of the present study was to determine if IBDV genomic sites, including the 5' end of vp5, could carry an influenza A virus hemagglutinin (HA) epitope. HA-expressing IBDVs were generated when the HA epitope was fused to the N-terminus of VP5 (HA5-IBDV) or VP4 (HA4-IBDV) or the C-terminus of VP1 (1HA-IBDV). Viral titers obtained after co-transfection with cDNA from the ha-containing segment and the complementary genomic segment were 1.3 × 104, 3.7 × 103 and 3.8 × 104 pfu/ml for HA5-IBDV, HA4-IBDV and 1HA-IBDV, respectively. The HA tag expression remained stable after 10 passages when the tag gene was inserted into the vp4 and vp1 genes. HA-IBDVs did not cause pathogenicity in specific-pathogen-free (SPF) chickens. However, only HA4-IBDV and 1HA-IBDV induced HA-specific antibodies, which were measured by ELISA with a maximum optical density (OD) value of 0.701 and 0.769, respectively, at 24 days after infection. Thus, IBDV can potentially be employed as a bivalent viral vector when the epitope is fused with VP4 or VP1.
机译:传染性法氏囊病病毒(IBDV)VP5的N端已被证明能够耐受小表位的插入。本研究的目的是确定IBDV基因组位点(包括vp5的5'端)是否可以携带A型流感病毒血凝素(HA)表位。当HA表位融合到VP5(HA5-IBDV)或VP4(HA4-IBDV)的N端或VP1(1HA-IBDV)的C端时,就会产生表达HA的IBDV。用HA含片段和互补基因组片段与cDNA共转染后获得的病毒滴度,对于HA5-IBDV,HA4-IBDV和1HA-IBDV分别为1.3×104、3.7×103和3.8×104 pfu / ml。将标签基因插入vp4和vp1基因后,经过10代,HA标签表达保持稳定。 HA-IBDV不会在无特定病原体(SPF)的鸡中引起致病性。但是,只有HA4-IBDV和1HA-IBDV可以诱导HA特异性抗体,这些抗体在感染后24天时通过ELISA测得的最大光密度(OD)值分别为0.701和0.769。因此,当表位与VP4或VP1融合时,IBDV可以潜在地用作二价病毒载体。

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