Monitoring and Modulation of Inducible Foxp3 + Regulatory T-Cell Differentiation in the Lymph Nodes Draining the Small Intestine and Colon

• Published in: Methods in molecular biology (Clifton, N.J.) Vol. 1559; p. 241
• Main Authors: Veenbergen, S, van Berkel, L A, du Pré, M F, Kozijn, A E, Samsom, Janneke N
• Format: Journal Article
• Language: English
• Published: United States 2017
• Subjects: Adjuvants, Immunologic - administration & dosage; Adoptive Transfer - methods; Animals; Biomarkers - metabolism; Cell Differentiation - immunology; Cell Lineage - immunology; Cholera Toxin - administration & dosage; Colon - cytology; Colon - immunology; Flow Cytometry; Forkhead Transcription Factors - genetics; Forkhead Transcription Factors - immunology; Gene Expression; Immunity, Mucosal; Intestine, Small - cytology; Intestine, Small - immunology; Lymph Nodes - cytology; Lymph Nodes - immunology; Mice; Mice, Inbred BALB C; Mice, Transgenic; Ovalbumin - administration & dosage; Receptors, Antigen, T-Cell - genetics; Receptors, Antigen, T-Cell - immunology; Staining and Labeling - methods; T-Lymphocytes, Regulatory - cytology; T-Lymphocytes, Regulatory - immunology; T-Lymphocytes, Regulatory - transplantation; Tregs; Mucosal immune regulation; Clonal competition; Small intestine and colon; iTregs; Foxp3; OT-II TCR transgenic; Adoptive transfer
• DOI: 10.1007/978-1-4939-6786-5_16

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.