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. 2014 Oct 21;1:20. doi: 10.3389/fvets.2014.00020

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Copyright © 2014 Roussey, Steibel and Coussens.

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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Treg expansion. PBMCs are isolated and used to generate MDMs by allowing adherence of monocytes to culture dishes followed by washing away of non-adherent cells. The adherent monocytes were allowed to mature into MDMs for 4 days followed by 24 h infection with MAP. After 3 days, autologous PBMCs are again isolated and FACS is used to select CD4⁺CD25⁺ lymphocytes. The cells are then combined with MAP-infected MDMs and cultured for 8 days with an enrichment cocktail, after which time the expanded CD4⁺CD25⁺ lymphocytes are ready for use in functional studies.