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Characterization of Monoclonal Antibodies against HA Protein of H1N1 Swine Influenza Virus and Protective Efficacy against H1 Viruses in Mice

机译:H1N1猪流感病毒HA蛋白单克隆抗体的表征及其对H1病毒的保护作用

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H1N1 swine influenza viruses (SIV) are prevalent in pigs globally, and occasionally emerge in humans, which raises concern about their pandemic threats. To stimulate hemagglutination (HA) of A/Swine/Guangdong/LM/2004 (H1N1) (SW/GD/04) antibody response, eukaryotic expression plasmid pCI-neo-HA was constructed and used as an immunogen to prepare monoclonal antibodies (mAbs). Five mAbs (designed 8C4, 8C6, 9D6, 8A4, and 8B1) against HA protein were obtained and characterized. Western blot showed that the 70 kDa HA protein could be detected by all mAbs in MDCK cells infected with SW/GD/04. Three mAbs—8C4, 8C6, and 9D6—have hemagglutination inhibition (HI) and neutralization test (NT) activities, and 8C6 induces the highest HI and NT titers. The protection efficacy of 8C6 was investigated in BALB/c mice challenged with homologous or heterologous strains of the H1 subtype SIV. The results indicate that mAb 8C6 protected the mice from viral infections, especially the homologous strain, which was clearly demonstrated by the body weight changes and reduction of viral load. Thus, our findings document for the first time that mAb 8C6 might be of potential therapeutic value for H1 subtype SIV infection.
机译:H1N1猪流感病毒(SIV)在全球的猪中普遍存在,并偶尔在人类中出现,这引起了人们对其大流行性威胁的担忧。为了刺激A /猪/广东/ LM / 2004(H1N1)(SW / GD / 04)抗体的血凝反应(HA),构建了真核表达质粒pCI-neo-HA,并用作免疫原来制备单克隆抗体(mAbs) )。获得并鉴定了针对HA蛋白的五种mAb(设计为8C4、8C6、9D6、8A4和8B1)。 Western印迹显示,被SW / GD / 04感染的MDCK细胞中的所有mAb均可检测到70 kDa HA蛋白。三种mAb(8C4、8C6和9D6)具有血凝抑制(HI)和中和测试(NT)活性,而8C6诱导出最高的HI和NT效价。在用H1亚型SIV的同源或异源菌株攻击的BALB / c小鼠中研究了8C6的保护功效。结果表明,mAb 8C6可以保护小鼠免受病毒感染,尤其是同源菌株的感染,这可以通过体重变化和病毒载量的降低得到明确证明。因此,我们的发现首次证明了mAb 8C6可能对H1亚型SIV感染具有潜在的治疗价值。

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