Anti-CD3 preconditioning reduced CD103⁺ DCs in MLN and reduced MLN DC capacity to induce donor T-cell expression of α4β7 receptor and CCR9. Spleen and MLN cells of BALB/c mice with or without anti-CD3 preconditioning were harvested and enriched for CD11c⁺ DCs by micromagnetic beads. The CD11c⁺-enriched cells were further analyzed with flow cytometry or used for in vitro culture. (A) A representative FACS pattern of CD103 expression among CD11c⁺ DCs. (B) Mean plus or minus SE of CD103⁺ cells among CD11c⁺ DCs and the yield of CD103⁺CD11c⁺ DCs in spleen and MLNs of 4 mice with or without anti-CD3 preconditioning. (C) Sorted CD4⁺/CD8⁺ T cells (0.2 × 10⁶) from C57BL/6 spleen were cocultured with enriched CD11c⁺ DCs (0.1 × 10⁶) from the MLNs of host BALB/c mice with or without anti-CD3 preconditioning for 4 days. Thereafter, donor CD4⁺ or CD8⁺ T cells were analyzed for the expression of α4β7 receptor and CCR9. One representative of 4 replicate experiments is shown. (D) Mean plus or minus SE of the percentage of α4β7⁺ or CCR9⁺ cells among donor CD4⁺ or CD8⁺ T cells in the culture of the 4 experiments. (E) Sorted donor CD8⁺ T cells (0.2 × 10⁶) were cocultured with CD103⁺ DCs (0.05 × 10⁶) from MLN and spleen of the host mice, and then donor CD8⁺ T-cell expression of α4β7 receptor and CCR9 were analyzed. The α4β7⁺ or CCR9⁺ CD8⁺ T cells were gated. One representative of 4 replicate experiments is shown. (F) Mean plus or minus SE of the α4β7⁺ receptor or CCR9⁺ cells among donor CD8⁺ T cells of the 4 experiments.