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S1 domain of the porcine epidemic diarrhea virus spike protein as a vaccine antigen

BACKGROUND: Porcine epidemic diarrhea virus (PEDV) is a highly contagious virus infecting pigs of all ages with high morbidity and mortality among newborn piglets. Currently, there is no effective vaccine available to protect the pigs from PEDV. The N-terminal subunit of spike protein (S1) is respon...

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Autores principales: Makadiya, Niraj, Brownlie, Robert, van den Hurk, Jan, Berube, Nathalie, Allan, Brenda, Gerdts, Volker, Zakhartchouk, Alexander
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4818391/
https://www.ncbi.nlm.nih.gov/pubmed/27036203
http://dx.doi.org/10.1186/s12985-016-0512-8
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author Makadiya, Niraj
Brownlie, Robert
van den Hurk, Jan
Berube, Nathalie
Allan, Brenda
Gerdts, Volker
Zakhartchouk, Alexander
author_facet Makadiya, Niraj
Brownlie, Robert
van den Hurk, Jan
Berube, Nathalie
Allan, Brenda
Gerdts, Volker
Zakhartchouk, Alexander
author_sort Makadiya, Niraj
collection PubMed
description BACKGROUND: Porcine epidemic diarrhea virus (PEDV) is a highly contagious virus infecting pigs of all ages with high morbidity and mortality among newborn piglets. Currently, there is no effective vaccine available to protect the pigs from PEDV. The N-terminal subunit of spike protein (S1) is responsible for virus binding to the cellular receptor and contains a number of neutralizing antibody epitopes. Thus, we expressed and produced recombinant S1 protein to protect newborn piglets by immunization of sows. METHODS: Affinity tagged PEDV S1 protein was expressed in a secretory form in yeast, insect and mammalian cells to identify the most suitable production system. Purified recombinant protein was analysed by SDS-PAGE, Western blot and deglycosylation assay. A pregnant sow was intramuscularly immunized three times with adjuvanted recombinant protein prior to farrowing. PEDV-specific immune responses in sera and colostrum of the sow and piglets were assayed by ELISA and virus neutralization assays. Piglets were challenged orally with PEDV, and clinical parameters were monitored for 6 days post-challenge. RESULTS AND CONCLUSION: Of three eukaryotic expression systems tested (yeast, insect-cell, and mammalian), expression by HEK-293 T cells gave the highest yield of protein that was N-glycosylated and was the most appropriate candidate for vaccination. Administration of the subunit vaccine in a sow resulted in induction of S1-specific IgG and IgA that were passively transferred to the suckling piglets. Also, high virus neutralization titres were observed in the serum of the vaccinated sow and its piglets. After PEDV challenge, piglets born to the vaccinated sow exhibited less severe signs of disease and significantly lower mortality compared to the piglets of a control sow. However, there were no significant differences in diarrhea, body weight and virus shedding. Thus, vaccination with S1 subunit vaccine failed to provide complete protection to suckling piglets after challenge exposure, and further improvements are needed for the development of a subunit vaccine that fully protects against PEDV infection.
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spelling pubmed-48183912016-04-03 S1 domain of the porcine epidemic diarrhea virus spike protein as a vaccine antigen Makadiya, Niraj Brownlie, Robert van den Hurk, Jan Berube, Nathalie Allan, Brenda Gerdts, Volker Zakhartchouk, Alexander Virol J Research BACKGROUND: Porcine epidemic diarrhea virus (PEDV) is a highly contagious virus infecting pigs of all ages with high morbidity and mortality among newborn piglets. Currently, there is no effective vaccine available to protect the pigs from PEDV. The N-terminal subunit of spike protein (S1) is responsible for virus binding to the cellular receptor and contains a number of neutralizing antibody epitopes. Thus, we expressed and produced recombinant S1 protein to protect newborn piglets by immunization of sows. METHODS: Affinity tagged PEDV S1 protein was expressed in a secretory form in yeast, insect and mammalian cells to identify the most suitable production system. Purified recombinant protein was analysed by SDS-PAGE, Western blot and deglycosylation assay. A pregnant sow was intramuscularly immunized three times with adjuvanted recombinant protein prior to farrowing. PEDV-specific immune responses in sera and colostrum of the sow and piglets were assayed by ELISA and virus neutralization assays. Piglets were challenged orally with PEDV, and clinical parameters were monitored for 6 days post-challenge. RESULTS AND CONCLUSION: Of three eukaryotic expression systems tested (yeast, insect-cell, and mammalian), expression by HEK-293 T cells gave the highest yield of protein that was N-glycosylated and was the most appropriate candidate for vaccination. Administration of the subunit vaccine in a sow resulted in induction of S1-specific IgG and IgA that were passively transferred to the suckling piglets. Also, high virus neutralization titres were observed in the serum of the vaccinated sow and its piglets. After PEDV challenge, piglets born to the vaccinated sow exhibited less severe signs of disease and significantly lower mortality compared to the piglets of a control sow. However, there were no significant differences in diarrhea, body weight and virus shedding. Thus, vaccination with S1 subunit vaccine failed to provide complete protection to suckling piglets after challenge exposure, and further improvements are needed for the development of a subunit vaccine that fully protects against PEDV infection. BioMed Central 2016-04-01 /pmc/articles/PMC4818391/ /pubmed/27036203 http://dx.doi.org/10.1186/s12985-016-0512-8 Text en © Makadiya et al. 2016 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research
Makadiya, Niraj
Brownlie, Robert
van den Hurk, Jan
Berube, Nathalie
Allan, Brenda
Gerdts, Volker
Zakhartchouk, Alexander
S1 domain of the porcine epidemic diarrhea virus spike protein as a vaccine antigen
title S1 domain of the porcine epidemic diarrhea virus spike protein as a vaccine antigen
title_full S1 domain of the porcine epidemic diarrhea virus spike protein as a vaccine antigen
title_fullStr S1 domain of the porcine epidemic diarrhea virus spike protein as a vaccine antigen
title_full_unstemmed S1 domain of the porcine epidemic diarrhea virus spike protein as a vaccine antigen
title_short S1 domain of the porcine epidemic diarrhea virus spike protein as a vaccine antigen
title_sort s1 domain of the porcine epidemic diarrhea virus spike protein as a vaccine antigen
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4818391/
https://www.ncbi.nlm.nih.gov/pubmed/27036203
http://dx.doi.org/10.1186/s12985-016-0512-8
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