Temperature Sensitivity: A Potential Method for the Generation of Vaccines against the Avian Coronavirus Infectious Bronchitis Virus
The <i>Gammacoronavirus</i> infectious bronchitis virus (IBV) is a highly contagious economically important respiratory pathogen of domestic fowl. Reverse genetics allows for the molecular study of pathogenic determinants to enable rational vaccine design. The recombinant IBV (rIBV) Beau...
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| Format: | Article |
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MDPI AG
2020-07-01
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| Series: | Viruses |
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| Online Access: | https://www.mdpi.com/1999-4915/12/7/754 |
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| author | Sarah Keep Phoebe Stevenson-Leggett Angela Steyn Michael S. Oade Isobel Webb Jamie Stuart Lonneke Vervelde Paul Britton Helena J. Maier Erica Bickerton |
| author_facet | Sarah Keep Phoebe Stevenson-Leggett Angela Steyn Michael S. Oade Isobel Webb Jamie Stuart Lonneke Vervelde Paul Britton Helena J. Maier Erica Bickerton |
| author_sort | Sarah Keep |
| collection | DOAJ |
| description | The <i>Gammacoronavirus</i> infectious bronchitis virus (IBV) is a highly contagious economically important respiratory pathogen of domestic fowl. Reverse genetics allows for the molecular study of pathogenic determinants to enable rational vaccine design. The recombinant IBV (rIBV) Beau-R, a molecular clone of the apathogenic Beaudette strain, has previously been investigated as a vaccine platform. To determine tissues in which Beau-R could effectively deliver antigenic genes, an in vivo study in chickens, the natural host, was used to compare the pattern of viral dissemination of Beau-R to the pathogenic strain M41-CK. Replication of Beau-R was found to be restricted to soft tissue within the beak, whereas M41-CK was detected in beak tissue, trachea and eyelid up to seven days post infection. In vitro assays further identified that, unlike M41-CK, Beau-R could not replicate at 41 °C, the core body temperature of a chicken, but is able to replicate a 37 °C, a temperature relatable to the very upper respiratory tract. Using a panel of rIBVs with defined mutations in the structural and accessory genes, viral replication at permissive and non-permissive temperatures was investigated, identifying that the Beau-R replicase gene was a determinant of temperature sensitivity and that sub-genomic mRNA synthesis had been affected. The identification of temperature sensitive allelic lesions within the Beau-R replicase gene opens up the possibility of using this method of attenuation in other IBV strains for future vaccine development as well as a method to investigate the functions of the IBV replicase proteins. |
| first_indexed | 2024-03-10T18:30:05Z |
| format | Article |
| id | doaj.art-e1ebe16b8a834077b186ccfe50678ec8 |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-03-10T18:30:05Z |
| publishDate | 2020-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj.art-e1ebe16b8a834077b186ccfe50678ec82023-11-20T06:42:46ZengMDPI AGViruses1999-49152020-07-0112775410.3390/v12070754Temperature Sensitivity: A Potential Method for the Generation of Vaccines against the Avian Coronavirus Infectious Bronchitis VirusSarah Keep0Phoebe Stevenson-Leggett1Angela Steyn2Michael S. Oade3Isobel Webb4Jamie Stuart5Lonneke Vervelde6Paul Britton7Helena J. Maier8Erica Bickerton9The Pirbright Institute, Pirbright, Surrey GU24 0NF, UKThe Pirbright Institute, Pirbright, Surrey GU24 0NF, UKThe Pirbright Institute, Pirbright, Surrey GU24 0NF, UKThe Pirbright Institute, Pirbright, Surrey GU24 0NF, UKThe Pirbright Institute, Pirbright, Surrey GU24 0NF, UKThe Pirbright Institute, Pirbright, Surrey GU24 0NF, UKDivision of Infection and Immunity, The Roslin Institute and Royal (Dick), School of Veterinary Studies, University of Edinburgh, Easter Bush, Midlothian EH25 9RG, UKThe Pirbright Institute, Pirbright, Surrey GU24 0NF, UKThe Pirbright Institute, Pirbright, Surrey GU24 0NF, UKThe Pirbright Institute, Pirbright, Surrey GU24 0NF, UKThe <i>Gammacoronavirus</i> infectious bronchitis virus (IBV) is a highly contagious economically important respiratory pathogen of domestic fowl. Reverse genetics allows for the molecular study of pathogenic determinants to enable rational vaccine design. The recombinant IBV (rIBV) Beau-R, a molecular clone of the apathogenic Beaudette strain, has previously been investigated as a vaccine platform. To determine tissues in which Beau-R could effectively deliver antigenic genes, an in vivo study in chickens, the natural host, was used to compare the pattern of viral dissemination of Beau-R to the pathogenic strain M41-CK. Replication of Beau-R was found to be restricted to soft tissue within the beak, whereas M41-CK was detected in beak tissue, trachea and eyelid up to seven days post infection. In vitro assays further identified that, unlike M41-CK, Beau-R could not replicate at 41 °C, the core body temperature of a chicken, but is able to replicate a 37 °C, a temperature relatable to the very upper respiratory tract. Using a panel of rIBVs with defined mutations in the structural and accessory genes, viral replication at permissive and non-permissive temperatures was investigated, identifying that the Beau-R replicase gene was a determinant of temperature sensitivity and that sub-genomic mRNA synthesis had been affected. The identification of temperature sensitive allelic lesions within the Beau-R replicase gene opens up the possibility of using this method of attenuation in other IBV strains for future vaccine development as well as a method to investigate the functions of the IBV replicase proteins.https://www.mdpi.com/1999-4915/12/7/754coronavirusIBVtemperature sensitivityreplicaseRNA synthesis |
| spellingShingle | Sarah Keep Phoebe Stevenson-Leggett Angela Steyn Michael S. Oade Isobel Webb Jamie Stuart Lonneke Vervelde Paul Britton Helena J. Maier Erica Bickerton Temperature Sensitivity: A Potential Method for the Generation of Vaccines against the Avian Coronavirus Infectious Bronchitis Virus Viruses coronavirus IBV temperature sensitivity replicase RNA synthesis |
| title | Temperature Sensitivity: A Potential Method for the Generation of Vaccines against the Avian Coronavirus Infectious Bronchitis Virus |
| title_full | Temperature Sensitivity: A Potential Method for the Generation of Vaccines against the Avian Coronavirus Infectious Bronchitis Virus |
| title_fullStr | Temperature Sensitivity: A Potential Method for the Generation of Vaccines against the Avian Coronavirus Infectious Bronchitis Virus |
| title_full_unstemmed | Temperature Sensitivity: A Potential Method for the Generation of Vaccines against the Avian Coronavirus Infectious Bronchitis Virus |
| title_short | Temperature Sensitivity: A Potential Method for the Generation of Vaccines against the Avian Coronavirus Infectious Bronchitis Virus |
| title_sort | temperature sensitivity a potential method for the generation of vaccines against the avian coronavirus infectious bronchitis virus |
| topic | coronavirus IBV temperature sensitivity replicase RNA synthesis |
| url | https://www.mdpi.com/1999-4915/12/7/754 |
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