Purification and Characterisation of Badger IgA and Its Detection in the Context of Tuberculosis
1
Department of Bacteriology, Animal and Plant Health Agency, Addlestone, Surrey KT15 3NB, UK
2
Laboratoire de la Rage et de la Faune Sauvage de Nancy (LRFSN), ANSES, 54220 Malzéville, France
3
Public Health England, Porton Down, Salisbury SP4 0JG, UK
4
School of Veterinary Medicine, Faculty of Health and Medical Sciences, University of Surrey, Guildford GU2 7AL, UK
*
Author to whom correspondence should be addressed.
Vet. Sci. 2019, 6(4), 89; https://doi.org/10.3390/vetsci6040089
Received: 30 September 2019 / Revised: 28 October 2019 / Accepted: 31 October 2019 / Published: 2 November 2019
(This article belongs to the Special Issue Mycobacterial Diseases in Animals)
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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Keywords:
immunoglobulin A; badger; tuberculosis
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MDPI and ACS Style
Dalley, D.; Lesellier, S.; Salguero, F.J.; Chambers, M.A. Purification and Characterisation of Badger IgA and Its Detection in the Context of Tuberculosis. Vet. Sci. 2019, 6, 89.
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