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A pilot study evaluating the utility of commercially available antibodies for flow cytometric analysis of Panthera species lymphocytes

BACKGROUND: The immune response against tuberculosis in lions is still poorly defined and our understanding is hampered by the lack of lion specific reagents. The process for producing antibodies against a specific antigen is laborious and not available to many research laboratories. As the search f...

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Autores principales: Sylvester, Tashnica Taime, Parsons, Sven David Charles, van Helden, Paul David, Miller, Michele Ann, Loxton, Andre Gareth
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6299994/
https://www.ncbi.nlm.nih.gov/pubmed/30567560
http://dx.doi.org/10.1186/s12917-018-1717-4
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author Sylvester, Tashnica Taime
Parsons, Sven David Charles
van Helden, Paul David
Miller, Michele Ann
Loxton, Andre Gareth
author_facet Sylvester, Tashnica Taime
Parsons, Sven David Charles
van Helden, Paul David
Miller, Michele Ann
Loxton, Andre Gareth
author_sort Sylvester, Tashnica Taime
collection PubMed
description BACKGROUND: The immune response against tuberculosis in lions is still poorly defined and our understanding is hampered by the lack of lion specific reagents. The process for producing antibodies against a specific antigen is laborious and not available to many research laboratories. As the search for antibody cross-reactivity is an important strategy for immunological studies in veterinary medicine, we have investigated the use of commercially available antibodies to characterize T cell subsets in African lions (Panthera leo). RESULTS: Commercially available antibodies were screened and investigated the influence of two different sample processing methods, as well as the effect of time delay on cell surface marker expression on lion lymphocytes. Using commercially available antibodies, we were able to identify CD4(+), CD5(+), CD8(+), CD14(+), CD25(+), CD44(+) and CD45(+) T lymphocytes in samples obtained by density gradient centrifugation as well as red cell lysis of lion whole blood. Two distinct lymphocyte populations, which differed in size and phenotype, were observed in the samples processed by density gradient centrifugation. CONCLUSION: Commercially available antibodies are able to differentiate between T lymphocyte subsets including immune effector cells in African lion whole blood, and possibly give insight into unique specie phenotypes. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12917-018-1717-4) contains supplementary material, which is available to authorized users.
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spelling pubmed-62999942018-12-20 A pilot study evaluating the utility of commercially available antibodies for flow cytometric analysis of Panthera species lymphocytes Sylvester, Tashnica Taime Parsons, Sven David Charles van Helden, Paul David Miller, Michele Ann Loxton, Andre Gareth BMC Vet Res Research Article BACKGROUND: The immune response against tuberculosis in lions is still poorly defined and our understanding is hampered by the lack of lion specific reagents. The process for producing antibodies against a specific antigen is laborious and not available to many research laboratories. As the search for antibody cross-reactivity is an important strategy for immunological studies in veterinary medicine, we have investigated the use of commercially available antibodies to characterize T cell subsets in African lions (Panthera leo). RESULTS: Commercially available antibodies were screened and investigated the influence of two different sample processing methods, as well as the effect of time delay on cell surface marker expression on lion lymphocytes. Using commercially available antibodies, we were able to identify CD4(+), CD5(+), CD8(+), CD14(+), CD25(+), CD44(+) and CD45(+) T lymphocytes in samples obtained by density gradient centrifugation as well as red cell lysis of lion whole blood. Two distinct lymphocyte populations, which differed in size and phenotype, were observed in the samples processed by density gradient centrifugation. CONCLUSION: Commercially available antibodies are able to differentiate between T lymphocyte subsets including immune effector cells in African lion whole blood, and possibly give insight into unique specie phenotypes. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12917-018-1717-4) contains supplementary material, which is available to authorized users. BioMed Central 2018-12-19 /pmc/articles/PMC6299994/ /pubmed/30567560 http://dx.doi.org/10.1186/s12917-018-1717-4 Text en © The Author(s). 2018 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 Article
Sylvester, Tashnica Taime
Parsons, Sven David Charles
van Helden, Paul David
Miller, Michele Ann
Loxton, Andre Gareth
A pilot study evaluating the utility of commercially available antibodies for flow cytometric analysis of Panthera species lymphocytes
title A pilot study evaluating the utility of commercially available antibodies for flow cytometric analysis of Panthera species lymphocytes
title_full A pilot study evaluating the utility of commercially available antibodies for flow cytometric analysis of Panthera species lymphocytes
title_fullStr A pilot study evaluating the utility of commercially available antibodies for flow cytometric analysis of Panthera species lymphocytes
title_full_unstemmed A pilot study evaluating the utility of commercially available antibodies for flow cytometric analysis of Panthera species lymphocytes
title_short A pilot study evaluating the utility of commercially available antibodies for flow cytometric analysis of Panthera species lymphocytes
title_sort pilot study evaluating the utility of commercially available antibodies for flow cytometric analysis of panthera species lymphocytes
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6299994/
https://www.ncbi.nlm.nih.gov/pubmed/30567560
http://dx.doi.org/10.1186/s12917-018-1717-4
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