Evaluation of a toxoid fusion protein vaccine produced in plants to protect poultry against necrotic enteritis

Joseph G.L. Hunter, Shyra Wilde, Amanda M. Tafoya, Jamie Horsman, Miranda Yousif, Andrew G. Diamos, Kenneth L. Roland, Hugh Mason

Research output: Contribution to journalArticle

Abstract

Background: Necrotic enteritis (NE) is caused by type A strains of the bacterium Clostridium perfringens. Total global economic losses to the poultry industry due to NE is estimated to be over two billion dollars annually. Traditionally, NE has been effectively controlled by inclusion of antibiotics in the diet of poultry. However, recent concerns regarding the impact of this practice on increasing antibiotic resistance in human pathogens have led us to consider alternative approaches, such as vaccination, for controlling this disease. NE strains of C. perfringens produce two major toxins, a-toxin and NetB. Immune responses against either toxin can provide partial protection against NE. Methods: We have developed a fusion protein combining a non-toxic carboxylterminal domain of a-toxin (PlcC) and an attenuated, mutant form of NetB (NetB-W262A) for use as a vaccine antigen to immunize poultry against NE. We utilized a DNA sequence that was codon-optimized for Nicotiana benthamiana to enable high levels of expression. The 6-His tagged PlcC-NetB fusion protein was synthesized in N. benthamiana using a geminiviral replicon transient expression system, purified by metal affinity chromatography, and used to immunize broiler birds. Results: Immunized birds produced a strong serum IgY response against both the plant produced PlcC-NetB protein and against bacterially produced His-PlcC and His-NetB. Immunized birds were significantly protected against a subsequent in-feed challenge with virulent C. perfringens when treated with the fusion protein. These results indicate that a plant-produced PlcC-NetB toxoid is a promising vaccine candidate for controlling NE in poultry.

Original languageEnglish (US)
Article numbere6600
JournalPeerJ
Volume2019
Issue number3
DOIs
StatePublished - Jan 1 2019
Externally publishedYes

Fingerprint

Toxoids
necrotic enteritis
Poultry
toxoids
Enteritis
poultry
Birds
Fusion reactions
Vaccines
vaccines
His-His-His-His-His-His
Clostridium perfringens
toxins
Proteins
proteins
Anti-Bacterial Agents
Nicotiana benthamiana
Affinity chromatography
Clostridium
Replicon

Keywords

  • Clostridium perfringens
  • Necrotic enteritis vaccine
  • Plant-based antigen production
  • Poultry vaccine

ASJC Scopus subject areas

  • Neuroscience(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Hunter, J. G. L., Wilde, S., Tafoya, A. M., Horsman, J., Yousif, M., Diamos, A. G., ... Mason, H. (2019). Evaluation of a toxoid fusion protein vaccine produced in plants to protect poultry against necrotic enteritis. PeerJ, 2019(3), [e6600]. https://doi.org/10.7717/peerj.6600

Evaluation of a toxoid fusion protein vaccine produced in plants to protect poultry against necrotic enteritis. / Hunter, Joseph G.L.; Wilde, Shyra; Tafoya, Amanda M.; Horsman, Jamie; Yousif, Miranda; Diamos, Andrew G.; Roland, Kenneth L.; Mason, Hugh.

In: PeerJ, Vol. 2019, No. 3, e6600, 01.01.2019.

Research output: Contribution to journalArticle

Hunter, JGL, Wilde, S, Tafoya, AM, Horsman, J, Yousif, M, Diamos, AG, Roland, KL & Mason, H 2019, 'Evaluation of a toxoid fusion protein vaccine produced in plants to protect poultry against necrotic enteritis' PeerJ, vol. 2019, no. 3, e6600. https://doi.org/10.7717/peerj.6600
Hunter JGL, Wilde S, Tafoya AM, Horsman J, Yousif M, Diamos AG et al. Evaluation of a toxoid fusion protein vaccine produced in plants to protect poultry against necrotic enteritis. PeerJ. 2019 Jan 1;2019(3). e6600. https://doi.org/10.7717/peerj.6600
Hunter, Joseph G.L. ; Wilde, Shyra ; Tafoya, Amanda M. ; Horsman, Jamie ; Yousif, Miranda ; Diamos, Andrew G. ; Roland, Kenneth L. ; Mason, Hugh. / Evaluation of a toxoid fusion protein vaccine produced in plants to protect poultry against necrotic enteritis. In: PeerJ. 2019 ; Vol. 2019, No. 3.
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abstract = "Background: Necrotic enteritis (NE) is caused by type A strains of the bacterium Clostridium perfringens. Total global economic losses to the poultry industry due to NE is estimated to be over two billion dollars annually. Traditionally, NE has been effectively controlled by inclusion of antibiotics in the diet of poultry. However, recent concerns regarding the impact of this practice on increasing antibiotic resistance in human pathogens have led us to consider alternative approaches, such as vaccination, for controlling this disease. NE strains of C. perfringens produce two major toxins, a-toxin and NetB. Immune responses against either toxin can provide partial protection against NE. Methods: We have developed a fusion protein combining a non-toxic carboxylterminal domain of a-toxin (PlcC) and an attenuated, mutant form of NetB (NetB-W262A) for use as a vaccine antigen to immunize poultry against NE. We utilized a DNA sequence that was codon-optimized for Nicotiana benthamiana to enable high levels of expression. The 6-His tagged PlcC-NetB fusion protein was synthesized in N. benthamiana using a geminiviral replicon transient expression system, purified by metal affinity chromatography, and used to immunize broiler birds. Results: Immunized birds produced a strong serum IgY response against both the plant produced PlcC-NetB protein and against bacterially produced His-PlcC and His-NetB. Immunized birds were significantly protected against a subsequent in-feed challenge with virulent C. perfringens when treated with the fusion protein. These results indicate that a plant-produced PlcC-NetB toxoid is a promising vaccine candidate for controlling NE in poultry.",
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