Abstract
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.
Highlights
Eurasian H1N1 swine influenza virus (SIV) was first reported in pigs in 1979 [1] and circulated in the European pig population [2]
DNA-based immunization can solve these problems because native proteins can be expressed in vivo when they are delivered in the form of DNA as an immunogen, which does not require the process of protein production or purification
The cytopathic effect (CPE) was observed daily and the number of wells showing more than 50% pathological changes were recorded
Summary
Eurasian H1N1 swine influenza virus (SIV) was first reported in pigs in 1979 [1] and circulated in the European pig population [2]. Vaccinations may not be effective in preventing against diverse viral strains, manifesting as less immunogenic, or acting with inadequate speed, to combat newly-emerging seasonal or potentially pandemic strains [16]. Studies have proved that mAbs could be used as an effective and preventive treatment against influenza virus infection [23,24,25,26,27]. Until now, there are no effective neutralizing mAbs available in preventing or controlling H1N1 SIV infection. DNA-based immunization can solve these problems because native proteins can be expressed in vivo when they are delivered in the form of DNA as an immunogen, which does not require the process of protein production or purification. We prepared and characterized five mAbs and evaluated 8C6 protective efficacy in mice against infection with homologous and heterologous H1 subtype viruses
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