Grouping Pig-Specific Responses to Mitogen with Similar Responder Animals may Facilitate the Interpretation of Results Obtained in an Out-Bred Animal Model

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Date
2014-04Author
Wilson, Heather
Pasternak, J. Alex
Ng, Siew Hon
Kaeser, Tobias
Meurens, Francois
Publisher
J Vacc VaccinolType
ArticlePeer Reviewed Status
Peer ReviewedMetadata
Show full item recordAbstract
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.
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: .Subject
Pig
PBMC
lymphocyte
mitogen
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.Collections
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