In vitro Differentiation of Thymic Treg Cell Progenitors to Mature Thymic Treg Cells

David L Owen; Michael A Farrar

doi:10.21769/BioProtoc.3335

In vitro Differentiation of Thymic T_reg Cell Progenitors to Mature Thymic T_reg Cells

Bio Protoc. 2019 Aug 20;9(16):e3335. doi: 10.21769/BioProtoc.3335.

Authors

David L Owen¹, Michael A Farrar¹

Affiliation

¹ Center for Immunology, Masonic Cancer Center, and the Department of Lab Medicine and Pathology, University of Minnesota, Minneapolis, MN, USA.

Abstract

Thymic T_reg cell differentiation occurs via a two-step process. Step one generates T_reg cell progenitors (T_regP) and is driven by strong TCR interactions with antigens presented in the thymus. Step two is initiated by activation of STAT5 via IL-2, or IL-15, leading to mature T_reg cells capable of emigrating from the thymus and mediating immune tolerance and homeostasis in peripheral tissues. Herein we describe an in vitro T_regP cell differentiation assay that models the second, cytokine dependent, step of thymic T_reg cell development. It can be utilized with relative ease to determine if a population of thymocytes represents a potential progenitor population for T_reg cells as well as test how different cytokines or chemical inhibitors modulate the differentiation of known T_regP cell populations into mature T_reg cells.

Keywords: Regulatory T (Treg) cells; Treg cell development; Cytokine stimulation; Foxp3; STAT5; T cell tolerance; Thymic selection; Thymus.