Figure 4.
Figure 4. pDCs recruited to LNs during EAE establish Ag-specific interactions with CD4+ T cells. (A) WT and pIII+IV−/− mice were injected with CFA and dLNs were harvested after 2 d. dLN sections from untreated and CFA-treated mice were stained with antibodies against PNAd to label HEVs and PDCA-1 to label pDCs. Histograms show the density (percentage of surface) of pDCs in regions situated near to (<20 µm) or distant from (>20 µm) HEVs. Results are representative of three independent experiments. The means and SEM derived from three independent mice per group are shown. ns, not significant; *, P < 0.05; **, P < 0.01. 7–12 confocal images were analyzed for each genotype and region. Representative images are shown. (B) CMTMR-labeled 2D2 CD4+ T cells were adoptively transferred into WT and pIII+IV−/− mice that had been injected 1 d previously with MOG35–55+CFA or CFA alone. dLNs were harvested 1 d after the T cell transfer, and sections were stained with antibodies against PDCA-1. Histograms show the percentages of 2D2 T cells that interact with pDCs and show the mean and SEM derived from at least two independent experiments. ns, not significant; *, P < 0.05; ***, P < 0.001. The numbers of confocal images analyzed (N) are indicated at the bottom of each bar. The ratios above the bars represent the number of interacting 2D2 T cells over the total number of 2D2 T cells. Representative images are shown for MOG35–55+CFA-treated WT and pIII+IV−/− mice. Arrowheads indicate 2D2 T cells that interact with pDCs. (C and D) CMTMR-labeled 2D2 or OT-II T cells were adoptively transferred into WT:WT and pIII+IV−/−:WT chimeras that had been immunized 1 d previously with MOG35–55+CFA. dLNs were harvested 1 d after the T cell transfer, and sections were stained with antibodies against PDCA-1. (C) The histogram shows the percentages of 2D2 or OT-II T cell that interact with pDCs and shows the mean and SEM derived from at least two independent experiments. ***, P < 0.001; ns, not significant. The numbers of confocal images analyzed (N) are indicated at the bottom of each bar. The ratios above the bars represent the number of interacting CD4+ T cells over the total number of CD4+ T cells. Representative images from WT:WT and pIII+IV−/−:WT chimeras are shown. Arrowheads indicate 2D2 T cells interacting with pDCs. (D) The histogram shows the distribution of pDCs in contact with one, two, three, or four 2D2 T cells. The numbers of pDCs that were analyzed are indicated above the bars. A representative image of a pDC interacting with three different 2D2 T cells in the LN of a WT:WT chimera is shown.

pDCs recruited to LNs during EAE establish Ag-specific interactions with CD4+ T cells. (A) WT and pIII+IV−/− mice were injected with CFA and dLNs were harvested after 2 d. dLN sections from untreated and CFA-treated mice were stained with antibodies against PNAd to label HEVs and PDCA-1 to label pDCs. Histograms show the density (percentage of surface) of pDCs in regions situated near to (<20 µm) or distant from (>20 µm) HEVs. Results are representative of three independent experiments. The means and SEM derived from three independent mice per group are shown. ns, not significant; *, P < 0.05; **, P < 0.01. 7–12 confocal images were analyzed for each genotype and region. Representative images are shown. (B) CMTMR-labeled 2D2 CD4+ T cells were adoptively transferred into WT and pIII+IV−/− mice that had been injected 1 d previously with MOG35–55+CFA or CFA alone. dLNs were harvested 1 d after the T cell transfer, and sections were stained with antibodies against PDCA-1. Histograms show the percentages of 2D2 T cells that interact with pDCs and show the mean and SEM derived from at least two independent experiments. ns, not significant; *, P < 0.05; ***, P < 0.001. The numbers of confocal images analyzed (N) are indicated at the bottom of each bar. The ratios above the bars represent the number of interacting 2D2 T cells over the total number of 2D2 T cells. Representative images are shown for MOG35–55+CFA-treated WT and pIII+IV−/− mice. Arrowheads indicate 2D2 T cells that interact with pDCs. (C and D) CMTMR-labeled 2D2 or OT-II T cells were adoptively transferred into WT:WT and pIII+IV−/−:WT chimeras that had been immunized 1 d previously with MOG35–55+CFA. dLNs were harvested 1 d after the T cell transfer, and sections were stained with antibodies against PDCA-1. (C) The histogram shows the percentages of 2D2 or OT-II T cell that interact with pDCs and shows the mean and SEM derived from at least two independent experiments. ***, P < 0.001; ns, not significant. The numbers of confocal images analyzed (N) are indicated at the bottom of each bar. The ratios above the bars represent the number of interacting CD4+ T cells over the total number of CD4+ T cells. Representative images from WT:WT and pIII+IV−/−:WT chimeras are shown. Arrowheads indicate 2D2 T cells interacting with pDCs. (D) The histogram shows the distribution of pDCs in contact with one, two, three, or four 2D2 T cells. The numbers of pDCs that were analyzed are indicated above the bars. A representative image of a pDC interacting with three different 2D2 T cells in the LN of a WT:WT chimera is shown.

This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal