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.

, , , , , , ,,
Methods in Molecular Biology
Department of Gastroenterology & Hepatology

Veenbergen, S., van Berkel, L., du Pré, F., Kozijn, A., & Samsom, J. (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