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. 2019 Mar 11;10:396. doi: 10.3389/fimmu.2019.00396

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Copyright © 2019 Rodríguez-Gómez, Talker, Käser, Stadler, Reiter, Ladinig, Milburn, Hammer, Mair, Saalmüller and Gerner.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Influence of aging on the expression of T-bet in blood-derived γδ T cells. (A) Following exclusion of dead cells (not shown), TCR-γδ⁺ T cells were gated within live lymphocytes (zebra plot) and further subgated to analyze the expression of T-bet. The scatter diagram shows the frequency of T-bet⁺ γδ T cells within live peripheral blood lymphocytes (PBL) isolated from pigs at 3 weeks, 25 weeks, 26 months, and 4 years of age. Each symbol represents data from one individual animal (for 3 weeks, 25 weeks, and 26 months of age animals (n = 5) were bled repeatedly; for 4 years of age, blood was obtained from different animals (n = 4); change in animals is indicated by a dashed line). Green bars indicate mean values. Data were subjected to Kruskal-Wallis test with Bonferroni correction as post-hoc test for pairwise comparison. (B) γδ T cells were gated for CD2⁺ and CD2⁻ subsets and further analyzed for co-expression of perforin and T-bet. Perforin⁺ and perforin⁻ T-bet⁺ γδ T cells were then gated and investigated for expression of CD27 and CD8α. Representative data for the four investigated stages of life are shown. Numbers located in gates and quadrants indicate percentage of cells for one particular phenotype. For T-bet/perforin-defined subpopulations below 10%, CD8α/CD27 expression is shown in dot-plots, for T-bet/perforin-defined subpopulations below 2%, CD8α/CD27 expression is not shown.