Protection of chickens against A/H5N1 virus by plant-based haemagglutinin fused IGMFC

Pham Thi Van, Ho Thi Huong, Nguyen Thu Giang, Pham Bich Ngoc, Vu Huyen Trang, Phan Trong Hoang, Tran Xuan Hanh, Udo Conrad, Chu Hoang Ha

Abstract


Vaccination is one of the most effective and cost-beneficial interventions for protection of animals against the highly pathogenic A/H5N1 avian influenza virus. Haemagglutinin (HA) is a transmembrane glycoprotein of A/H5N1 virus and is a critical antigen for development of the influenza vaccine. The haemagglutinin-based vaccine produced in plants was demonstrated as a candidate vaccine since it elicited neutralizing antibodies against A/H5N1 virus. In this study, immunogenicity and protective ability of a plant-based recombinant HA antigen which was fused to IgMFc to form oligomerized HA antigen (H5TG oligomer) had been evaluated by vaccination in chickens. Chicken sera after each vaccination were collected for Western blot, ELISA and HI assays. Ten days after the second vaccination, the chickens have been challenged with A/duck/TG/NAVET(3)/2013 virus, clade 1.1. The analysis results showed that the oligomeric recombinant H5TG antigen elicited stronger H5TG-specific IgY antibodies and A/H5N1 clade 1.1 virus-neutralizing antibodies than the H5TGpII trimeric recombinant antigen without fusing IgMFc in vaccinated chickens. Notably, the chicken protection rate against A/H5N1 clade 1.1 virus of the H5TG oligomer antigen was 80% that is not significantly lower than that of a commercial vaccine as a positive control from National Veterinary Joint Stock company NAVETCO, Vietnam with the chicken protective rate of 90%. Whereas the chicken protection rate against A/H5N1 clade 1.1 virus of the H5TG trimer antigen was 50%. These results suggest that the IgMFc motif plays an important role in the forming of oligomeric proteins which had been proved for enhancing immunogenicity and protection ability in this study. Therefore, the plant-based oligomerized recombinant H5TG antigen is a potential vaccine candidate against A/H5N1 influenza virus in the future.


Keywords


Haemagglutinin, recombinant antigen, vaccination, virus challenge, neutralizing antibodies, protective rate.

Full Text:

PDF

References


Floss DM, Conrad U (2012) Plant molecular pharming – veterinary applications. In: Meyers R A (Ed.): Encyclopedia of sustainability science and technology. Springer 11: 8073-8080.

Gerhard W (2001). The role of the antibody response in influenza virus infection. Curr Top Microbiol Immunol 260: 171-190.

Kalthoff D, Giritch A, Geisler K, Bettmann U, Klimyuk V, Hehnen HR, Gleba Y, Beer M (2010) Immunization with plant-expressed hemagglutinin protects chickens from lethal highly pathogenic avian influenza Virus H5N1 challenge infection. J Virol 84: 12002-12010.

Klenk HD (2015). Influenza Viruses En Route from Birds to Man. Cell Host Microb 15(6): 653-4.

O'Connor, T.P. SNAP Assay Technology. Top Companion Anim. Med. 2015, 30, 132-8. DOI: 10.1053/j.tcam.2015.12.002.

Office International des Epizooties (OIE) - World Organization for Animal Health. Highly pathogenic avian influenza, in manual of diagnostic tests and vaccines for terrestrial animals (mammals, birds, and bees). 2004, pp. 258–269. http://www.oie.int/en/international-standard-setting/terrestrial-manual/access-online/.

Office International des Epizooties (OIE) - World Organization for Animal Health. Update on avian influenza in animals. 2019. http://www.oie.int/en/animal-health-in-the-world/update-on-avian-influenza/.

Pham NB, Ho TT, Nguyen GT, Le TT, Le NT, Chang HC, Pham MD, Conrad U, Chu HH (2017) Nanodiamond enhances immune responses in mice against recombinant HA/H7N9 protein. J. nanobiotechnology 15: 69, doi:10.1186/s12951-017-0305-2.

Phan HT, Floss DM, Conrad U (2013) “Veterinary vaccines from transgenic plants: highlights of two decades of research and a promising example”. Curr. Pharm. Design 19: 5601-5611.

Phan HT (2012) ELPylated avian flu vaccines from plants: Improvement of expression and development of a new purification strategy. Thesis of PhD, Martin-Luther-Universität Halle-Wittenberg, Halle (Saale), 02 October 2012.

Phan HT, Pohl J, Floss DM, Rabenstein F, Veits J, Le BT, Chu HH, Hause G, Mettenleiter T, Conrad U (2013) ELPylated haemagglutinins produced in tobacco plants induce potentially neutralizing antibodies against H5N1 viruses in mice. Plant Biotechnol J 11: 582–593, doi:10.1111/pbi.12049.

Shoji Y, Bi H, Musiychuk K, Rhee A, Horsey A, Roy G, Green B, Shamloul M, Farrance CE, and Taggart B (2009). Plant-derived hemagglutinin protects ferrets against challenge infection with the A/Indonesia/05/05 strain of avian influenza. Vaccine 27: 1087-1092.

World Health Organization (WHO). Cumulative number of confirmed human cases for avian influenza A(H5N1) reported to WHO, 2003-2019. https://www.who.int/influenza/human_animal_interface/2019_04_09_tableH5N1.pdf?ua=1.




DOI: https://doi.org/10.15625/1811-4989/18/3/14483 Display counter: Abstract : 142 views. PDF : 96 views.