Figure 5. CD103⁺ DCs from neonatally SEA treated mice exhibit enhanced gut-imprinting and FoxP3-inducing ability in vitro.

Mice (n = 6 in each group) were fed staphylococcal enterotoxin A (SEA) or PBS (SHAM) perorally on six occasions during the first two weeks of life. Mesenteric lymph nodes (MLNs) were collected at four weeks after treatment and CD103⁺CD11c⁺and CD103^negCD11c⁺cells were separated by CD11c-positive MACS isolation followed by sorting for CD103⁺ cells on a flow cytometer. The DCs were pulsed with OVA and co-cultured with OVA-specific DO11.10 CD4⁺ T cells in medium containing 100 nM retinol. FoxP3 and α4β7 expression was determined in KJ1.26⁺CD4⁺ T cells, by flow cytometry, after six days of culture. (A) Representative dot plots of unsorted and CD103-sorted CD11c⁺ DCs. (B) Representative dot plot and gating strategy of the CD4⁺KJ1.26⁺ T cells. C) Proportion of FoxP3^neg α4β7^neg, FoxP3⁺ α4β7⁺, FoxP3^neg α4β7⁺and FoxP3⁺ α4β7^neg cells among the CD4⁺KJ1.26⁺ T cells cultivated with CD103⁺DCs, CD103^neg DCs and CD103^negDCs in cultures without Retinol. Error bars represent SD and horizontal line shows the median value of the group. Data shown are representative of two independent experiments. * P<0.05, analyzed with Mann-Whitney U-test.