Abstract
Developing new vaccine candidates is considered the best strategy for protecting poultry against artificial haemagglutinin (A/H5N1) strains. The transient expression system in plants has been a very efficient method for rapidly producing haemagglutinin-based recombinant vaccines. In this study, two novel artificial trimeric haemagglutinin constructs representing A/H5N1 strains that were detected in poultry from 2005 to 2015 in Vietnam, H5.c1 (representing all of the subclades 1.1, 1.1.1, and 1.1.2) and H5.c2 (representing all of the subclades 2.3.2.1, 2.3.2.1a, 2.3.2.1b, and 2.3.2.1c), were designed for transient expression in Nicotiana benthamiana via agroinfiltration. However, only the H5.c1 protein, which showed the best expression and biofunction via the haemagglutination test, was selected for purification by immobilized metal ion affinity chromatography (IMAC). The trimeric structure of the IMAC-purified H5.c1 protein was well characterized by cross-linking reaction and size exclusion chromatography. An indirect ELISA and Western blot analysis of vaccinated mouse sera demonstrated that the H5.c1 protein strongly induced HA-specific Immunoglobulin G (IgG) immune responses. Notably, the H5.c1 protein induced strongly neutralizing antibodies against homologous H5.c1 protein and that of three heterologous native strains of clade, 1, 1.1, and 2.3.2.1c, in haemagglutination inhibition assays. Therefore, the plant-based artificial H5.c1 protein can be a promising vaccine candidate for conferring poultry resistance against A/H5N1 viruses in Vietnam.
Highlights
Asian highly pathogenic avian influenza (HPAI) A (H5N1) virus is potentially able to cause a global pandemic due to its relatively large spread via avian hosts and its potential to directly infect humans [1]
The complete HA sequences of 90, 38, 34, 19, 143, 28, and 165 of A/H5N1 subclades 1.1, 1.1.1, 1.1.2, 2.3.2.1, 2.3.2.1a, 2.3.2.1b, and 2.3.2.1c, respectively, that were detected in poultry from 2005 to 2015 in Vietnam were collected from GenBank, National Center for Biotechnology Information (NCBI) and Global Initiative on Sharing All Influenza Data (GISAID)
Four H5 sequences were submitted to http://raptorx.uchicago.edu/ to predict the structure of these proteins, including two sequences of the clade 1 group
Summary
Asian highly pathogenic avian influenza (HPAI) A (H5N1) virus is potentially able to cause a global pandemic due to its relatively large spread via avian hosts and its potential to directly infect humans [1]. In Vietnam, over 2750 outbreaks of H5N1 occurring in poultry were reported to the World. Organisation for Animal Health (OIE) from the end of 2003 to 03 July 2019, among which one outbreak killed and disposed of 1120 birds in 2019 [2]. The viral HA protein plays a critical role in the infectivity and pathogenicity of influenza virus by binding to. Sci. 2019, 9, 4605 sialic acid on glycoconjugates of host membrane proteins and enabling virus entry into host cells [3]
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