The mucosa-draining lymphoid tissue favors differentiation of inducible Foxp3+ regulatory T cells. Adoptive transfer of T-cell receptor (TCR) transgenic (Tg) T cells is a powerful tool to study antigen-specific regulatory T-cell differentiation in lymphoid tissues in vivo. The kinetics and nature of the T-cell response largely depend on the route of antigen administration and degree of clonal competition. Here, we describe that adoptive transfer of CD4+ DO11.10 TCR Tg T cells can be used for monitoring Foxp3+ regulatory T-cell differentiation in the gut-draining lymph nodes. We describe two routes of mucosal antigen administration, e.g., the oral and intracolonic route known to induce T-cell responses in the small intestine-draining mesenteric lymph nodes (MLN) and distal colon-draining caudal and iliac lymph nodes (ILN), respectively. In particular, we discuss differences in frequency of inducible Foxp3+ regulatory T cells after adoptive transfer of variable numbers of Tg T cells and various amounts of orally gavaged ovalbumin (OVA), and explain how Foxp3+ regulatory T-cell differentiation can be modulated by coadministration of the adjuvant cholera toxin (CT) with OVA using this adoptive transfer system.

Additional Metadata
Keywords Adoptive transfer, Clonal competition, Foxp3, ITregs, Mucosal immune regulation, OT-II TCR transgenic, Small intestine and colon, Tregs
Persistent URL,
Series Methods in Molecular Biology
Veenbergen, S, van Berkel, L.A, du Pré, M.F, Kozijn, A.E, & Samsom, J.N. (2017). Monitoring and modulation of inducible Foxp3+ regulatory T-cell differentiation in the lymph nodes draining the small intestine and colon. In Inflammation : Methods and Protocols (pp. 241–254). doi:10.1007/978-1-4939-6786-5_16