Figure 3

COX-2-dependent PGs are required for functional Tr-cell induction in the gut-draining lymphoid tissue. (a) BALB/c mice enriched with CD4⁺ KJ1.26⁺ × RAG^−/− T cells were tolerized as described in Figure 1. Representative dot plot of dividing Foxp3-positive T cells in the MLN at 72 h after OVA feed. (b) BALB/c mice enriched with CD4⁺ KJ1.26⁺ cells were treated with NS-398 (black symbols) or vehicle (white symbols). At 48 h after OVA administration, MLNs were isolated and CD4⁺ KJ1.26⁺ cells within single-cell suspensions were analyzed for the percentage of Foxp3⁺ cells by flow cytometry and Foxp3 mRNA (24 and 48 h). (c) At 48 h after feeding, MLN and PP were isolated and single-cell suspensions were enriched for CD4⁺ cells. A total of 5 × 10⁵ enriched CD4⁺ cells from either NS-398-treated mice (black circles: MLN ; black diamonds: PP cells) or vehicle-treated mice (white squares: MLN cells; PP cells: white diamonds) were transferred to naive BALB/c recipients. As a control, 5 × 10⁵ enriched CD4⁺ cells isolated from MLN of saline-treated mice as a DTH control (white triangles) were transferred to naive acceptor BALB/c mice. Acceptor mice were sensitized and challenged as described in Figure 1. *Statistically significant (P<0.05). COX-2, cyclooxygenase-2; MLN, mesenteric lymph node; OVA, ovalbumin; PG, prostaglandin; PP, Peyer's Patch;

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