Fig. 4: Intranasal immunization with CTB-T2544 directs DC and lymphocyte migration.

a Twelve days after the primary immunization, MLN cells were surface-stained with CD11c and analyzed by flow cytometry. b, c Gut homing receptor expression on B and T lymphocytes. Twelve days after the primary immunization, lymphocytes isolated from the MLN (b) and spleen (c) of the experimental mice were stained with fluorochrome-tagged antibodies against cell lineage markers (CD4 and B220) and gut homing receptors (α4β7 and CCR9) and analyzed by flow cytometry. Data presented as percentages of cells out of the total T and B cells, expressing the homing receptors. d RT-qPCR analysis of gene expressions in DCs, isolated from the MLN of experimental mice were plotted. GAPDH-normalized expression of all the genes was represented as fold changes of the PBS-immunized group. All experiments were repeated three times, and data from a representative one are presented in (a) and mean ( ± SD) of the data from all three experiments are shown under (b–d). Significance was calculated using one way ANOVA (a) and two-way ANOVA (b,c) with post hoc analysis using Tukey’s multiple comparison test. Error bar represents SD. *p = 0.0281, ***p = 0.0008 and ****p < 0.0001 (T2544 vs. CTB-T2544).