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Osteopontin expression is essential for interferon-α production by plasmacytoid dendritic cells


The observation that the T-bet transcription factor allows tissue-specific upregulation of intracellular osteopontin (Opn-i) in plasmacytoid dendritic cells (pDCs) suggests that Opn might contribute to the expression of interferon-α (IFN-α) in those cells. Here we show that Opn deficiency substantially reduced Toll-like receptor 9 (TLR9)–dependent IFN-α responses but spared expression of transcription factor NF-κB–dependent proinflammatory cytokines. Shortly after TLR9 engagement, colocalization of Opn-i and the adaptor molecule MyD88 was associated with induction of transcription factor IRF7–dependent IFN-α gene expression, whereas deficient expression of Opn-i was associated with defective nuclear translocation of IRF7 in pDCs. The importance of the Opn–IFN-α pathway was emphasized by its essential involvement in cross-presentation in vitro and in anti–herpes simplex virus 1 IFN-α response in vivo. The finding that Opn-i selectively coupled TLR9 signaling to expression of IFN-α but not to that of other proinflammatory cytokines provides new molecular insight into the biology of pDCs.

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We thank S. Turley for critical reading; D. Laznik for technical assistance; and A. Angel for assistance with the manuscript and figures. Supported by the National Institutes of Health (AI48125 and AI12184 to H.C.; T32 CA70083 to M.L.S; and CA48126 and AI56296 to L.H.G.), the Ellison Medical Foundation (L.H.G.) and the Juvenile Diabetes Research Foundation (M.L.S.).

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Correspondence to Harvey Cantor.

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Supplementary information

Supplementary Fig. 1

Activation marker expression on pDC from Opn WT and Opn-deficient mice. (PDF 82 kb)

Supplementary Fig. 2

In vitro cross-presentation by pDC and cDC. (PDF 50 kb)

Supplementary Fig. 3

IFNAR signaling licenses pDC for antigen cross-presentation. (PDF 51 kb)

Supplementary Fig. 4

Evaluation of Opn-s and Opn-i in antigen cross-presentation. (PDF 54 kb)

Supplementary Methods (PDF 15 kb)

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Figure 1: Opn expression in pDCs and cDCs 24 h after stimulation.
Figure 2: Cytokine production by pDCs.
Figure 3: Intracellular Opn is required for IFN-α production.
Figure 4: Biochemical analysis of Opn-i.
Figure 5: Localization of Opn together with MyD88 and TLR9.
Figure 6: In vitro OVA cross-presentation by pDCs.
Figure 7: IFN-α-dependent cross-presentation.
Figure 8: Opn-dependent in vivo response to HSV-1 infection.