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Purification and Characterisation of Badger IgA and Its Detection in the Context of Tuberculosis

European badgers are a wildlife reservoir of bovine tuberculosis in parts of Great Britain. Accurate diagnosis of tuberculosis in badgers is important for the development of strategies for the control of the disease. Sensitive serological tests for badger TB are needed for reasons such as cost and s...

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Autores principales: Dalley, Deanna, Lesellier, Sandrine, Salguero, Francisco J., Chambers, Mark A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6958344/
https://www.ncbi.nlm.nih.gov/pubmed/31684068
http://dx.doi.org/10.3390/vetsci6040089
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author Dalley, Deanna
Lesellier, Sandrine
Salguero, Francisco J.
Chambers, Mark A.
author_facet Dalley, Deanna
Lesellier, Sandrine
Salguero, Francisco J.
Chambers, Mark A.
author_sort Dalley, Deanna
collection PubMed
description European badgers are a wildlife reservoir of bovine tuberculosis in parts of Great Britain. Accurate diagnosis of tuberculosis in badgers is important for the development of strategies for the control of the disease. Sensitive serological tests for badger TB are needed for reasons such as cost and simplicity. Assay of mucosal IgA could be useful for diagnosing respiratory pathogens such as Mycobacterium bovis and for monitoring the response to mucosal vaccination. To develop an IgA assay, we purified secretory IgA from badger bile, identifying secretory component (SC), heavy chain (HC) and light chain (LC), at 66, 46 and 27 Kda, respectively, on the basis of size comparison with other species. Monoclonal antibodies (mAbs) were generated to purified IgA. We selected two for ELISA development. The detection limit of the IgA-specific mAbs was found to be approximately 20 ng/mL when titrated against purified badger bile. One monoclonal antibody specific for badger IgA was used to detect IgA in serum and tracheal aspirate with specificity to an immunodominant antigen of M. bovis. An M. bovis infection dose-dependent IgA response was observed in experimentally infected badgers. IgA was also detected by immunohistochemistry in the lungs of bTB-infected badgers. With further characterisation, these represent new reagents for the study of the IgA response in badgers.
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spelling pubmed-69583442020-01-23 Purification and Characterisation of Badger IgA and Its Detection in the Context of Tuberculosis Dalley, Deanna Lesellier, Sandrine Salguero, Francisco J. Chambers, Mark A. Vet Sci Article European badgers are a wildlife reservoir of bovine tuberculosis in parts of Great Britain. Accurate diagnosis of tuberculosis in badgers is important for the development of strategies for the control of the disease. Sensitive serological tests for badger TB are needed for reasons such as cost and simplicity. Assay of mucosal IgA could be useful for diagnosing respiratory pathogens such as Mycobacterium bovis and for monitoring the response to mucosal vaccination. To develop an IgA assay, we purified secretory IgA from badger bile, identifying secretory component (SC), heavy chain (HC) and light chain (LC), at 66, 46 and 27 Kda, respectively, on the basis of size comparison with other species. Monoclonal antibodies (mAbs) were generated to purified IgA. We selected two for ELISA development. The detection limit of the IgA-specific mAbs was found to be approximately 20 ng/mL when titrated against purified badger bile. One monoclonal antibody specific for badger IgA was used to detect IgA in serum and tracheal aspirate with specificity to an immunodominant antigen of M. bovis. An M. bovis infection dose-dependent IgA response was observed in experimentally infected badgers. IgA was also detected by immunohistochemistry in the lungs of bTB-infected badgers. With further characterisation, these represent new reagents for the study of the IgA response in badgers. MDPI 2019-11-02 /pmc/articles/PMC6958344/ /pubmed/31684068 http://dx.doi.org/10.3390/vetsci6040089 Text en © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Dalley, Deanna
Lesellier, Sandrine
Salguero, Francisco J.
Chambers, Mark A.
Purification and Characterisation of Badger IgA and Its Detection in the Context of Tuberculosis
title Purification and Characterisation of Badger IgA and Its Detection in the Context of Tuberculosis
title_full Purification and Characterisation of Badger IgA and Its Detection in the Context of Tuberculosis
title_fullStr Purification and Characterisation of Badger IgA and Its Detection in the Context of Tuberculosis
title_full_unstemmed Purification and Characterisation of Badger IgA and Its Detection in the Context of Tuberculosis
title_short Purification and Characterisation of Badger IgA and Its Detection in the Context of Tuberculosis
title_sort purification and characterisation of badger iga and its detection in the context of tuberculosis
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6958344/
https://www.ncbi.nlm.nih.gov/pubmed/31684068
http://dx.doi.org/10.3390/vetsci6040089
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