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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:对小鼠H1N1猪HA蛋白的单克隆抗体的表征及对小鼠H1病毒的保护效应

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摘要

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)在全球猪普遍存在,偶尔出现在人类中,这引起了对流行性威胁的关注。为了刺激A / Swine / Guangdong / LM / 2004(H1N1)(SW / GD / 04)抗体反应的血凝(HA),构建了真核表达质粒pCI-Neo-Ha并用作免疫原制备单克隆抗体(mAbs )。获得了对HA蛋白的5mAb(设计的8C4,8C​​6,9D6,8A4和8B1)。 Western印迹表明,MDCK细胞中的所有MAB可以检测到70kDa的HA蛋白在感染的MDCK细胞中检测到SW / GD / 04。三种MAb-8C4,8C​​6和9D6 - 具有血粒子凝集抑制(HI)和中和试验(NT)活性,8C6诱导最高HI和NT滴度。在H1亚型SIV的同源或异源菌株攻击的BALB / C小鼠中研究了8C6的保护效果。结果表明,MAB 8C6保护了来自病毒感染的小鼠,特别是同源菌株,其通过体重变化清楚地证明了病毒载荷。因此,我们的调查结果是第一次MAB 8C6可能具有H1亚型SIV感染的潜在治疗价值。

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