本文關(guān)鍵詞：歐洲型PRRSV重組腺病毒疫苗的構(gòu)建及免疫效果評價　出處：《中國病原生物學(xué)雜志》2015年11期 　論文類型：期刊論文

  更多相關(guān)文章： EU-PRRSV 重組腺病毒 構(gòu)建 免疫評價

【摘要】：目的構(gòu)建歐洲型PRRSV重組腺病毒疫苗并對其免疫原性進(jìn)行分析,為歐洲型PRRSV的感染預(yù)防提供依據(jù)。方法 PCR擴(kuò)增歐洲型藍(lán)耳病病毒ORF3、ORF5目的基因,將其連接到腺病毒穿梭載體,構(gòu)建重組腺病毒穿梭質(zhì)粒pacad-EU-ORF3、pacad-EU-ORF5、pacad-EU-ORF3-ORF5。將重組腺病毒穿梭質(zhì)粒與骨架質(zhì)粒共同轉(zhuǎn)染至HEK 293細(xì)胞,構(gòu)建重組腺病毒,并對構(gòu)建的病毒用Western blot方法進(jìn)行驗(yàn)證。用構(gòu)建的重組腺病毒免疫BALB/c小鼠,通過淋巴細(xì)胞增殖試驗(yàn)對重組疫苗免疫效果進(jìn)行初步評價。結(jié)果 Western blot顯示重組腺病毒可穩(wěn)定表達(dá)目的蛋白ORF3和ORF5,分子質(zhì)量單位分別為30ku和22ku,與預(yù)期值相符。免疫小鼠淋巴細(xì)胞增殖試驗(yàn)顯示重組腺病毒刺激指數(shù)均為PBS組1.5倍以上(P0.05),且rAd-EU-ORF3-ORF5組顯著高于rAd-EU-ORF3組和rAd-EU-ORF5組,表明重組腺病毒疫苗能夠誘導(dǎo)機(jī)體淋巴細(xì)胞增殖,增強(qiáng)機(jī)體的細(xì)胞免疫應(yīng)答水平。結(jié)論成功包裝出重組腺病毒rAdEU-ORF3、rAd-EU-ORF5和rAd-EU-ORF3-ORF5,動物實(shí)驗(yàn)顯示其具有良好的免疫原性。
[Abstract]:Objective to construct a European type PRRSV recombinant adenovirus vaccine and analyze its immunogenicity to provide a basis for the prevention of infection in European type PRRSV. Methods the target genes of ORF3 and ORF5 were amplified by PCR, and then connected to adenovirus shuttle vector. The Recombinant Adenovirus Shuttle Plasmid pacad-EU-ORF3, pacad-EU-ORF5 and pacad-EU-ORF3-ORF5 were constructed. The Recombinant Adenovirus Shuttle Plasmid and skeleton plasmid were co transfected into HEK 293 cells, and then the recombinant adenovirus was constructed, and the constructed virus was verified by Western blot method. The recombinant adenovirus was used to immunise BALB/c mice, and the immune effect of the recombinant vaccine was evaluated by lymphocyte proliferation test. Results Western blot showed that recombinant adenovirus could stabilize the expression of target protein ORF3 and ORF5, and the molecular weight units were 30ku and 22ku, which were consistent with the expected value. Immunized mice lymphocyte proliferation test showed that the recombinant adenovirus stimulation index was more than 1.5 times for the PBS group (P0.05), and rAd-EU-ORF3-ORF5 group was significantly higher than that of rAd-EU-ORF3 group and rAd-EU-ORF5 group, showed that the recombinant adenovirus vaccine can induce lymphocyte proliferation in the body, enhance the cellular immune response of organism. Conclusion the recombinant adenovirus rAdEU-ORF3, rAd-EU-ORF5 and rAd-EU-ORF3-ORF5 were successfully packaged, and the animal experiments showed that the recombinant adenovirus had good immunogenicity.
【作者單位】： 吉林農(nóng)業(yè)大學(xué)動物科學(xué)技術(shù)學(xué)院;軍事醫(yī)學(xué)科學(xué)院軍事獸醫(yī)研究所;廣西大學(xué)動物科學(xué)技術(shù)學(xué)院;
【基金】：國家863計(jì)劃項(xiàng)目(No.2011AA10A208) 吉林省科技廳項(xiàng)目(No.20140309024NY)
【分類號】：R392
【正文快照】： ***ORF5,and rAd-EU-ORF3-ORF5were successfully constructed.Animal experiments indicated that the recombinant ade-novirus vaccines had good immunogenicity.豬繁殖與呼吸綜合征(porcine reproductive andrespiratory syndrome,PRRS)是由豬繁殖與呼吸綜合征病毒(porci

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