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. 2017 Apr 13;7:46408. doi: 10.1038/srep46408

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Copyright © 2017, The Author(s)

This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

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PBMCs were collected from three protected piglets from each group on the 18^th dpc (n = 3). Spleen and mesenteric lymph node (MLN) were collected from the two surviving piglets of the unvaccinated control group, two randomly selected piglets of the PO group, and the two protected piglets of the IM group (n = 2), and processed at necropsy on the 33^th dpc. IL-2, IFN-γ or TNF-α positive T cells (CD3⁺) in PBMCs (A) were measured by flow cytometry after stimulation with inactivated S. Choleraesuis antigen and staining with labelled anti-pig antibodies. IFN-γ positive CD4⁺ and CD8⁺ T cells in spleen and MLN (B) were also measured.