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Front Immunol. 2017 May 17;8:565. doi: 10.3389/fimmu.2017.00565. eCollection 2017.

Pathogen-Reactive T Helper Cell Analysis in the Pig.

Author information

1
Department of Veterinary Medicine, Institute of Immunology, Freie Universität Berlin, Berlin, Germany.
2
Department of Veterinary Medicine, Institute of Animal Nutrition, Freie Universität Berlin, Berlin, Germany.
3
Faculty of Veterinary Medicine, Institute of Immunology, Centre for Infectious Diseases, University of Leipzig, Leipzig, Germany.
4
Faculty of Veterinary Medicine, Institute for Bacteriology and Mycology, Centre for Infectious Diseases, University of Leipzig, Leipzig, Germany.
5
Laboratory of Parasitology, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium.

Abstract

There is growing interest in studying host-pathogen interactions in human-relevant large animal models such as the pig. Despite the progress in developing immunological reagents for porcine T cell research, there is an urgent need to directly assess pathogen-specific T cells-an extremely rare population of cells, but of upmost importance in orchestrating the host immune response to a given pathogen. Here, we established that the activation marker CD154 (CD40L), known from human and mouse studies, identifies also porcine antigen-reactive CD4+ T lymphocytes. CD154 expression was upregulated early after antigen encounter and CD4+CD154+ antigen-reactive T cells coexpressed cytokines. Antigen-induced expansion and autologous restimulation enabled a time- and dose-resolved analysis of CD154 regulation and a significantly increased resolution in phenotypic profiling of antigen-responsive cells. CD154 expression identified T cells responding to staphylococcal Enterotoxin B superantigen stimulation as well as T cells responding to the fungus Candida albicans and T cells specific for a highly prevalent intestinal parasite, the nematode Ascaris suum during acute and trickle infection. Antigen-reactive T cells were further detected after immunization of pigs with a single recombinant bacterial antigen of Streptococcus suis only. Thus, our study offers new ways to study antigen-specific T lymphocytes in the pig and their contribution to host-pathogen interactions.

KEYWORDS:

Ascaris suum; CD154; CD40 ligand; Candida albicans; Streptococcus suis; antigen-specific; pig; porcine CD4 T cell

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