Grouping Pig-Specific Responses to Mitogen with Similar Responder Animals may Facilitate the Interpretation of Results Obtained in an Out-Bred Animal Model
Date
2014-04
Authors
Wilson, Heather
Pasternak, J. Alex
Ng, Siew Hon
Kaeser, Tobias
Meurens, Francois
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J Vacc Vaccinol
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Article
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Abstract
Pig peripheral blood-derived mononuclear cells (PBMCs) and lamina propria mononuclear cells (LPMCs) stimulated with mitogens ex vivo can show significant animal-to-animal variation lead to difficulty in interpreting responses in an out-bred animal species. Mixed-cell populations were stimulated ex vivo with 2.5 μg/ml Con A or 2.5 ng/ml PMA plus 250 ng/ml ionomycin (PMAi; (LPCMs only)) or media alone for 72 hours. Supernatants were then tested for cytokine production using a Bioplex assay for porcine IFNα, IFNγ, IL-10, and IL-12. Unstimulated PBMCs
had significant levels of IL-10 and the median value for this group decreased in the presence of Con A. Con A did,
however, induce production of IFNα and IFNγ, but not IL-12 in this cell population. In contrast, unstimulated and Con
A-stimulated LPMCs produced negligible IL-10, IFNα, IFNγ, and the majority of animals’ LPMCs showed negligible
IL-12 production in response to Con A. In contrast, LPMCs stimulated with PMAi produced IFNγ suggesting cytokine
production is mitogen–specific response. When we tracked animal-specific responses, we observed that discrete
subsets of animal’s PBMCs responded to Con A with significantly increased or decreased IL-10 production relative
to unstimulated cells. Further, in the LPMCs, some cells produced no IL-12 in response to Con A but showed
augmented production in response to PMAi, while others showed production of IL-12 in response to Con A but no
response to PMAi. Flow cytometric analysis showed that the PBMCs were a mixture of CD3+ T cells>CD21+ B cells>CD172+ myeloid cells whereas the LPMCs consisted of mainly Cytotoxic T cells and Natural Killer cells. The percentage of CD8α+CD4+ antigen-experienced T cells was greater in the LPMCs relative to the PBMCs. As expected in an out-bred species, animal-specific differences in cytokine production in response to stimulants exist and may confound interpretation of results unless tracked individually.
Description
Copyright: © 2014 J. Alex Pasternak, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Keywords
Pig, PBMC, lymphocyte, mitogen
Citation
[1] Pasternak JA, Ng S, Käser T, Meurens F, Wilson HL. Grouping Pig-Specific Responses to Mitogen with Similar Responder Animals may Facilitate the Interpretation of Results Obtained in an Out-Bred Animal Model. J Vacc Vaccinol. 2014;5: .
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DOI
dx.doi.org/10.4172/2157-7560.1000242