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

Abstract

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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References

  1. 1

    Iwasaki, A. & Medzhitov, R. Toll-like receptor control of the adaptive immune responses. Nat. Immunol. 5, 987–995 (2004).

  2. 2

    Liu, Y.J. IPC: professional type 1 interferon-producing cells and plasmacytoid dendritic cell precursors. Annu. Rev. Immunol. 23, 275–306 (2005).

  3. 3

    Krug, A. et al. TLR9-dependent recognition of MCMV by IPC and DC generates coordinated cytokine responses that activate antiviral NK cell function. Immunity 21, 107–119 (2004).

  4. 4

    Colonna, M., Trinchieri, G. & Liu, Y.J. Plasmacytoid dendritic cells in immunity. Nat. Immunol. 5, 1219–1226 (2004).

  5. 5

    Banchereau, J., Pascual, V. & Palucka, A.K. Autoimmunity through cytokine-induced dendritic cell activation. Immunity 20, 539–550 (2004).

  6. 6

    Theofilopoulos, A.N., Baccala, R., Beutler, B. & Kono, D.H. Type I interferons (α/β) in immunity and autoimmunity. Annu. Rev. Immunol. 23, 307–336 (2005).

  7. 7

    Le Bon, A. et al. Cross-priming of CD8+ T cells stimulated by virus-induced type I interferon. Nat. Immunol. 4, 1009–1015 (2003).

  8. 8

    Boonstra, A. et al. Flexibility of mouse classical and plasmacytoid-derived dendritic cells in directing T helper type 1 and 2 cell development: dependency on antigen dose and differential toll-like receptor ligation. J. Exp. Med. 197, 101–109 (2003).

  9. 9

    Salio, M., Palmowski, M.J., Atzberger, A., Hermans, I.F. & Cerundolo, V. CpG-matured murine plasmacytoid dendritic cells are capable of in vivo priming of functional CD8 T cell responses to endogenous but not exogenous antigens. J. Exp. Med. 199, 567–579 (2004).

  10. 10

    Honda, K., Yanai, H., Takaoka, A. & Taniguchi, T. Regulation of the type I IFN induction: a current view. Int. Immunol. 17, 1367–1378 (2005).

  11. 11

    Kawai, T. & Akira, S. Pathogen recognition with Toll-like receptors. Curr. Opin. Immunol. 17, 338–344 (2005).

  12. 12

    Honda, K. et al. Spatiotemporal regulation of MyD88–IRF7 signalling for robust type-I interferon induction. Nature 434, 1035–1040 (2005).

  13. 13

    Ashkar, S. et al. Eta-1 (osteopontin): an early component of Type 1 (cell-mediated) immunity. Science 287, 860–864 (2000).

  14. 14

    Shinohara, M.L. et al. T-bet-dependent expression of osteopontin contributes to T cell polarization. Proc. Natl. Acad. Sci. USA 102, 17101–17106 (2005).

  15. 15

    Miyazaki, T. et al. Implication of allelic polymorphism of osteopontin in the development of lupus nephritis in MRL/lpr mice. Eur. J. Immunol. 35, 1510–1520 (2005).

  16. 16

    Nau, G.J. et al. Attenuated host resistance against Mycobacterium bovis BCG infection in mice lacking osteopontin. Infect. Immun. 67, 4223–4230 (1999).

  17. 17

    Sibalic, V., Fan, X., Loffing, J. & Wuthrich, R.P. Upregulated renal tubular CD44, hyaluronan, and osteopontin in kdkd mice with interstitial nephritis. Nephrol. Dial. Transplant. 12, 1344–1353 (1997).

  18. 18

    Yu, X.Q. et al. A functional role for osteopontin in experimental crescentic glomerulonephritis in the rat. Proc. Assoc. Am. Phys. 110, 50–64 (1998).

  19. 19

    Hudkins, K.L. et al. Osteopontin expression in human crescentic glomerulonephritis. Kidney Int. 57, 105–116 (2000).

  20. 20

    Steinman, L., Martin, R., Bernard, C., Conlon, P. & Oksenberg, J.R. Multiple sclerosis: deeper understanding of its pathogenesis reveals new targets for therapy. Annu. Rev. Neurosci. 25, 491–505 (2002).

  21. 21

    Xu, G. et al. Role of osteopontin in amplification and perpetuation of rheumatoid synovitis. J. Clin. Invest. 115, 1060–1067 (2005).

  22. 22

    Comabella, M. et al. Plasma osteopontin levels in multiple sclerosis. J. Neuroimmunol. 158, 231–239 (2005).

  23. 23

    Gravallese, E.M. Osteopontin: a bridge between bone and the immune system. J. Clin. Invest. 112, 147–149 (2003).

  24. 24

    Zohar, R. et al. Intracellular osteopontin is an integral component of the CD44-ERM complex involved in cell migration. J. Cell. Physiol. 184, 118–130 (2000).

  25. 25

    Suzuki, K. et al. Colocalization of intracellular osteopontin with CD44 is associated with migration, cell fusion, and resorption in osteoclasts. J. Bone Miner. Res. 17, 1486–1497 (2002).

  26. 26

    Zhu, B. et al. Osteopontin modulates CD44-dependent chemotaxis of peritoneal macrophages through G-protein-coupled receptors: evidence of a role for an intracellular form of osteopontin. J. Cell. Physiol. 198, 155–167 (2004).

  27. 27

    Wilson, H.L. & O'Neill, H.C. Identification of differentially expressed genes representing dendritic cell precursors and their progeny. Blood 102, 1661–1669 (2003).

  28. 28

    Renkl, A.C. et al. Osteopontin functionally activates dendritic cells and induces their differentiation toward a Th1-polarizing phenotype. Blood 106, 946–955 (2005).

  29. 29

    Cantor, H. T-cell receptor crossreactivity and autoimmune disease. Adv. Immunol. 75, 209–233 (2000).

  30. 30

    Denhardt, D.T., Noda, M., O'Regan, A.W., Pavlin, D. & Berman, J.S. Osteopontin as a means to cope with environmental insults: regulation of inflammation, tissue remodeling, and cell survival. J. Clin. Invest. 107, 1055–1061 (2001).

  31. 31

    Diao, H. et al. Osteopontin as a mediator of NKT cell function in T cell-mediated liver diseases. Immunity 21, 539–550 (2004).

  32. 32

    Weiss, J.M. et al. Osteopontin is involved in the initiation of cutaneous contact hypersensitivity by inducing Langerhans and dendritic cell migration to lymph nodes. J. Exp. Med. 194, 1219–1230 (2001).

  33. 33

    Szabo, S.J. et al. A novel transcription factor, T-bet, directs Th1 lineage commitment. Cell 100, 655–669 (2000).

  34. 34

    Lugo-Villarino, G., Ito, S., Klinman, D.M. & Glimcher, L.H. The adjuvant activity of CpG DNA requires T-bet expression in dendritic cells. Proc. Natl. Acad. Sci. USA 102, 13248–13253 (2005).

  35. 35

    Lugo-Villarino, G., Maldonado-Lopez, R., Possemato, R., Penaranda, C. & Glimcher, L.H. T-bet is required for optimal production of IFN-γ and antigen-specific T cell activation by dendritic cells. Proc. Natl. Acad. Sci. USA 100, 7749–7754 (2003).

  36. 36

    Hemmi, H., Kaisho, T., Takeda, K. & Akira, S. The roles of Toll-like receptor 9, MyD88, and DNA-dependent protein kinase catalytic subunit in the effects of two distinct CpG DNAs on dendritic cell subsets. J. Immunol. 170, 3059–3064 (2003).

  37. 37

    Lee, S.W. et al. Effects of a hexameric deoxyriboguanosine run conjugation into CpG oligodeoxynucleotides on their immunostimulatory potentials. J. Immunol. 165, 3631–3639 (2000).

  38. 38

    Durand, V., Wong, S.Y., Tough, D.F. & Le Bon, A. Shaping of adaptive immune responses to soluble proteins by TLR agonists: a role for IFN-α/β. Immunol. Cell Biol. 82, 596–602 (2004).

  39. 39

    Haeryfar, S.M. The importance of being a pDC in antiviral immunity: the IFN mission versus Ag presentation? Trends Immunol. 26, 311–317 (2005).

  40. 40

    Zuniga, E.I., McGavern, D.B., Pruneda-Paz, J.L., Teng, C. & Oldstone, M.B. Bone marrow plasmacytoid dendritic cells can differentiate into myeloid dendritic cells upon virus infection. Nat. Immunol. 5, 1227–1234 (2004).

  41. 41

    Baccala, R., Kono, D.H. & Theofilopoulos, A.N. Interferons as pathogenic effectors in autoimmunity. Immunol. Rev. 204, 9–26 (2005).

  42. 42

    Ballas, Z.K. et al. Divergent therapeutic and immunologic effects of oligodeoxynucleotides with distinct CpG motifs. J. Immunol. 167, 4878–4886 (2001).

  43. 43

    Bonifacino, J.S. & Traub, L.M. Signals for sorting of transmembrane proteins to endosomes and lysosomes. Annu. Rev. Biochem. 72, 395–447 (2003).

  44. 44

    Honda, K. et al. Role of a transductional-transcriptional processor complex involving MyD88 and IRF7 in Toll-like receptor signaling. Proc. Natl. Acad. Sci. USA 101, 15416–15421 (2004).

  45. 45

    Takaoka, A. et al. Integral role of IRF-5 in the gene induction programme activated by Toll-like receptors. Nature 434, 243–249 (2005).

  46. 46

    Cho, H.J. et al. IFN-αβ promote priming of antigen-specific CD8+ and CD4+ T lymphocytes by immunostimulatory DNA-based vaccines. J. Immunol. 168, 4907–4913 (2002).

  47. 47

    Datta, S.K. et al. A subset of Toll-like receptor ligands induces cross-presentation by bone marrow-derived dendritic cells. J. Immunol. 170, 4102–4110 (2003).

  48. 48

    Heit, A. et al. Cutting edge: Toll-like receptor 9 expression is not required for CpG DNA-aided cross-presentation of DNA-conjugated antigens but essential for cross-priming of CD8 T cells. J. Immunol. 170, 2802–2805 (2003).

  49. 49

    Barchet, W. et al. Dendritic cells respond to influenza virus through TLR7- and PKR-independent pathways. Eur. J. Immunol. 35, 236–242 (2005).

  50. 50

    Abel, B., Freigang, S., Bachmann, M.F., Boschert, U. & Kopf, M. Osteopontin is not required for the development of Th1 responses and viral immunity. J. Immunol. 175, 6006–6013 (2005).

  51. 51

    Hron, J.D. & Peng, S.L. Type I IFN protects against murine lupus. J. Immunol. 173, 2134–2142 (2004).

  52. 52

    Li, J. et al. Deficiency of type I interferon contributes to Sle2-associated component lupus phenotypes. Arthritis Rheum. 52, 3063–3072 (2005).

  53. 53

    Baechler, E.C. et al. Interferon-inducible gene expression signature in peripheral blood cells of patients with severe lupus. Proc. Natl. Acad. Sci. USA 100, 2610–2615 (2003).

  54. 54

    Kono, D.H., Baccala, R. & Theofilopoulos, A.N. Inhibition of lupus by genetic alteration of the interferon-α/β receptor. Autoimmunity 36, 503–510 (2003).

  55. 55

    Rittling, S.R. et al. Mice lacking osteopontin show normal development and bone structure but display altered osteoclast formation in vitro. J. Bone Miner. Res. 13, 1101–1111 (1998).

  56. 56

    Nakano, H., Yanagita, M. & Gunn, M.D. CD11c+B220+Gr-1+ cells in mouse lymph nodes and spleen display characteristics of plasmacytoid dendritic cells. J. Exp. Med. 194, 1171–1178 (2001).

  57. 57

    McCarty, N. et al. Signaling by the kinase MINK is essential in the negative selection of autoreactive thymocytes. Nat. Immunol. 6, 65–72 (2005).

  58. 58

    Gilliet, M. & Liu, Y.J. Generation of human CD8 T regulatory cells by CD40 ligand-activated plasmacytoid dendritic cells. J. Exp. Med. 195, 695–704 (2002).

  59. 59

    Hu, D. et al. Analysis of regulatory CD8 T cells in Qa-1-deficient mice. Nat. Immunol. 5, 516–523 (2004).

  60. 60

    Andrews, N.C. & Faller, D.V. A rapid micropreparation technique for extraction of DNA-binding proteins from limiting numbers of mammalian cells. Nucleic Acids Res. 19, 2499 (1991).

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Acknowledgements

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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The authors declare no competing financial interests.

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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Further reading

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.