Figure 1.
Frequency of Foxp3+ T-regs in cervical lymph node is increased in a Myd88/TICAM-1 independent manner.
(A) Cervical lymph nodes (CLN), axillary lymph nodes (ALN), inguinal lymph nodes (ILN), and mesenteric lymph nodes (MLN) from wild type B6 mice were analyzed for the expression of Foxp3. The isotype control is shown at the bottom. Plots were gated on CD4+ T cells. Representative of seven separate experiments. (B) As in (A), but CLN, ALN, ILN and MLN from wild-type B6 mice (WT) or Myd88/TICAM1 double knockout mice (Myd88/TICAM1 DKO) were analyzed for the expression of Foxp3. Summary from seven separate experiments. P value provided is by paired t-test or t-test. “n.s.” = “not significant”. (C) As in (B), but a representative of CLN is shown.
Figure 2.
Frequency of ROR-γt+CD4+ T cell in cervical lymph node is increased in a Myd88/TICAM-1 dependent manner.
(A) CLN, ALN, ILN, MLN, and spleen (Sp) from WT B6 mice were analyzed for the expression of Foxp3 and ROR-γt. The plots were gated on CD4+ T cells. Isotype staining for ROR-γt is shown at the bottom. Representative of three separate experiments. (B) As in (A), but cells from WT mice or Myd88/TICAM-1 DKO mice were analyzed for the expression of ROR-γt and CD4. The graphic shows a summary from two separate experiments. P value provided is by paired-t test or t-test. “n.s.” = “not significant”.
Figure 3.
Cervical lymph nodes are draining lymph nodes from the oral cavity.
CFSE-labeled OTII CD4+T cells were adoptively transferred into B6 mice on day −1. On day 0, 500 µg of OVA was administered sublingually (s.l.). CLN or ALN was analyzed for CFSE dilution at day 3. One of two similar experiments is shown for the FACS plots. Plots were gated on CD4+T cells.
Figure 4.
Dendritic cells from oral-cavity-draining cervical lymph nodes induce Foxp3+ T-regs.
(A) CLNs were stained with Foxp3 (red), CD4 (green) and CD11c (blue). Representative of three similar separate experiments. (B) OT II CD4+ T cells (5×104) were cultured with dendritic cells (DCs) from CLN, ALN, or MLN (5×104) with or without OVA peptide. After 5 days, cells were stained with Foxp3, CD25 and CD4. The plots were gated on CD4+ T cells. Representative of four separate experiments. (C) As in (A), but the graphic shows a summary of four separate experiments. P value provided is by paired t-test. “n.s.” = “not significant”.
Figure 5.
Phenotype of dendritic cells from cervical lymph nodes.
(A) DCs from CLN, ALN, and MLN were freshly prepared from B6 mice. mRNA was prepared and real-time PCR was performed. Expression of each sample was normalized to GAPDH mRNA expression and fold increase of each sample was calculated relative to the expression at 0 h. One of two separate experiments is shown. (B) DCs from CLN, ALN, and MLN were analyzed for the expression of CD103. The plots were gated on CD11c+ cells. The isotype control for CD103 is shown at the bottom. The graphic shows a summary of four separate experiments. Average +/− SD is shown. (C) As in (B), DCs from CLN, ALN, and MLN were analyzed for the expression of MHC class II or B220. The plots were gated on CD11c+ cells. The graphic shows a summary of four separate experiments. Average +/− SD is shown. (D) As in (B), DCs from CLN, ALN, and MLN were analyzed for the expression of CD8. The plots were gated on CD11c+ cells. The graphic shows a summary of 10 separate experiments. P value provided is by paired t-test. “n.s.” = “not significant”.