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The Cholera Toxin B Subunit (CTB) Fused to the Porcine Arterivirus Matrix M and GP5 Envelope Proteins Fails to Enhance the GP5-Specific Antibody Response in Pigs Immunized with Adenovectors

Abstract

The porcine reproductive and respiratory syndrome virus (PRRSV) is an arterivirus of the Arteriviridae family. As the current commercial vaccines are incompletely protective effective against PRRSV infection, we developed a vaccine strategy using replicating but non-disseminating adenovectors (rAdVs) expressing the PRRSV M matrix protein in fusion with the neutralizing major epitope-carrying GP5 envelope protein (Roques et al. in Vet Res 44:17, 2013). Although production of GP5-specific antibodies (Abs) was observed, no PRRSV-specific neutralizing Abs (NAbs) were induced in pigs given the rAdVs expressing M-GP5 or M-GP5m (GP5m being a mutant form of GP5). Nevertheless, partial protection was observed in the M-GP5m-rAdV-inoculated pigs experimentally infected with PRRSV. Here, we determined the impact of the cholera toxin B subunit (CTB, known for its adjuvant effect) in fusion with the C-terminus of M-GP5m on the Ab response to PRRSV. Three-week-old pigs were immunized twice both intramuscularly and intranasally at 3-week intervals with rAdV-expressing the green fluorescent protein (rAdV-GFP), rAdV-M-GP5m, or rAdV-M-GP5m-CTB. Pigs immunized with rAdV-M-GP5m showed a high level of serum GP5-specific Abs (as determined by an indirect ELISA). In contrast, CTB in fusion with M-GP5m had an unexpected severe negative impact on GP5-specific Ab production. PRRSV-specific NAbs could not be detected in any pigs of all groups.

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Acknowledgments

This work was supported by grants from the Natural Sciences and Engineering Research Council of Canada (NSERC, strategic program), Agriculture and Agri-Food Canada (AAC), the “Conseil pour le Dévelopement de l’Agriculture du Québec” (CDAQ), the “Fédération des Producteurs de Porcs du Québec” (FPPQ), and “les Conseils d’Adaptation du porc de l’Alberta, du Manitoba et de la Saskatchewan”. E. Roques was supported by graduate student fellowships from the “Centre de Recherche en Infectiologie Porcine et Avicole” (CRIPA) and “La Fondation UQAM”. The authors thank Marie-Claude St-Louis, Frédéric Beaudoin, Nedzad Music, Wilfried Saron, and Aurélie Girard for technical assistance and/or helpful discussion, Steve Méthot for statistical analyses of the data, and all employees who took care of the animals during the project.

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Correspondence to Denis Archambault.

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Roques, E., Lessard, M. & Archambault, D. The Cholera Toxin B Subunit (CTB) Fused to the Porcine Arterivirus Matrix M and GP5 Envelope Proteins Fails to Enhance the GP5-Specific Antibody Response in Pigs Immunized with Adenovectors. Mol Biotechnol 57, 701–708 (2015). https://doi.org/10.1007/s12033-015-9861-6

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  • DOI: https://doi.org/10.1007/s12033-015-9861-6

Keywords

  • CTB
  • PRRSV
  • GP5 and M proteins
  • Adenovectors
  • Pigs
  • Antibody response