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. 2016 Oct 17;7:421. doi: 10.3389/fimmu.2016.00421

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Copyright © 2016 Maggioli, Palmer, Thacker, Vordermeier, McGill, Whelan, Larsen, Jacobs and Waters.

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) or licensor 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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Specific Tcm responses to vaccination at 6 weeks post-vaccination. For long-term culture, PBMCs were isolated at 6 WPV and stimulated with a cocktail of Ag85A, TB10.4, and PPD for 13 days followed by transfer to 96-well round bottom plates with APCs and addition of media alone, PPD, Ag85A/TB10.4, or ESAT-6:CFP10 for an additional 16 h. Cytokine production and effector/memory phenotype were determined by ICS flow cytometry. Results are presented as average and SE. Polyfunctional responses by Tcm cells (gated on CD45RO⁺/CCR7⁺/CD4⁺ cells) were detected upon recall stimulation with PPD (A), Ag85A/TB10.4 (B), but not to ESAT-6:CFP10 (C). *Different (P < 0.05, Tukey’s) responses between non-vaccinates and both vaccinated groups.