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. 2008 Aug 20;105(34):12439–12444. doi: 10.1073/pnas.0806439105

Fig. 2.

Fig. 2.

pDCs in the PLNs produce IFN-α. (A) Homogenates were prepared from the PLNs of 2-, 3-, 4-, 6-, and 12-week-old NOD mice and used for IFN-α ELISA. Data were obtained from three independent experiments, each pooled from 5 mice. (B and C) Single cell suspensions were prepared from the PLNs, ILNs, and spleen of 2-, 4-, and 6-week-old NOD mice (n = 3 per group), stained with mAbs against CD11c and mPDCA1, and examined by FACS. Data for the frequency of pDCs (B; CD11cintmPDCA-1+) and CD11chigh mDCs (C; CD11chighmPDCA-1−) were obtained from 3–4 mice at each time point. (D) Single cell suspensions were prepared from the PLNs of 10-day-old (blue) and 3-week-old (red) NOD mice and stained with mAbs against mPDCA-1, CD11c, and IFN-α. The overlay histogram shows the intracellular IFN-α level on gated CD11intmPDCA1+ pDCs. (E) Frozen sections of 4-week-old NOD mouse PLNs were stained with mAbs against mPDCA1 and CD3 (left) or mAbs against mPDCA-1 and CD19 (right) and visualized on a fluorescent microscope. Most pDCs (red) were localized in the CD3+ T cell area (green), whereas very few pDCs (red) were found in the CD19+ B cell area (green). The isotype control mAb for anti-CD3, CD19, or mPDCA1 mAb did not stain any cells (data not shown). These staining results are representative of those from 4 mice.