Article | Published:

Siglec-G is a B1 cell–inhibitory receptor that controls expansion and calcium signaling of the B1 cell population

Nature Immunology volume 8, pages 695704 (2007) | Download Citation

Abstract

B1 cells are an important cell population for the production of natural antibodies and for antibacterial immunoglobulin responses. Here we identified the mouse protein Siglec-G as a B1 cell inhibitory receptor. Siglec-G was expressed in a B cell–restricted way, with large amounts present in B1 cells. When overexpressed, Siglec-G inhibited B cell receptor–mediated calcium signaling. Siglec-G-deficient mice had massive expansion of the B1a cell population, which began early in development and was B cell intrinsic. Siglec-G-deficient mice had higher titers of natural IgM antibodies but not a higher penetrance of IgG autoantibodies. Siglec-G-deficient B1 cells showed a strongly enhanced calcium signaling. Our results demonstrate that Siglec-G-dependent negative regulation exists in B1 cells, which may explain the naturally muted signaling response of B1 cells.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Accessions

Primary accessions

GenBank/EMBL/DDBJ

References

  1. 1.

    & Origins and functions of B-1 cells with notes on the role of CD5. Annu. Rev. Immunol. 20, 253–300 (2002).

  2. 2.

    & Marginal-zone B cells. Nat. Rev. Immunol. 2, 323–335 (2002).

  3. 3.

    , , & B-1a and B-1b cells exhibit distinct developmental requirements and have unique functional roles in innate and adaptive immunity to S. pneumoniae. Immunity 23, 7–18 (2005).

  4. 4.

    et al. B1b lymphocytes confer T cell-independent long-lasting immunity. Immunity 21, 379–390 (2004).

  5. 5.

    et al. Regulation of B1 cell migration by signals through Toll-like receptors. J. Exp. Med. 203, 2541–2550 (2006).

  6. 6.

    B-1 cells: the lineage question revisited. Immunol. Rev. 175, 9–22 (2000).

  7. 7.

    , & Identification of a B-1 B cell-specified progenitor. Nat. Immunol. 7, 293–301 (2006).

  8. 8.

    , & Treatment of murine CD5- B cells with anti-Ig, but not LPS, induces surface CD5: two B-cell activation pathways. Int. Immunol. 3, 467–476 (1991).

  9. 9.

    & B cell antigen receptor specificity and surface density together determine B-1 versus B-2 cell development. J. Exp. Med. 190, 471–477 (1999).

  10. 10.

    , , , & Production of immunoglobulin isotypes by Ly-1+ B cells in viable motheaten and normal mice. Science 232, 1423–1425 (1986).

  11. 11.

    , & CD72-deficient mice reveal nonredundant roles of CD72 in B cell development and activation. Immunity 11, 495–506 (1999).

  12. 12.

    et al. CD22 regulates thymus-independent responses and the lifespan of B cells. Nature 384, 634–637 (1996).

  13. 13.

    , , , & CD22 is a negative regulator of B-cell receptor signalling. Curr. Biol. 7, 133–143 (1997).

  14. 14.

    , , & Hyperresponsive B cells in CD22-deficient mice. Science 274, 798–801 (1996).

  15. 15.

    et al. CD22 is both a positive and negative regulator of B lymphocyte antigen receptor signal transduction: altered signaling in CD22-deficient mice. Immunity 5, 551–562 (1996).

  16. 16.

    , , & Negative regulation of antigen receptor-mediated signaling by constitutive association of CD5 with the SHP-1 protein tyrosine phosphatase in B-1 B cells. Eur. J. Immunol. 29, 3319–3328 (1999).

  17. 17.

    , , , & CD5-mediated negative regulation of antigen receptor-induced growth signals in B-1 B cells. Science 274, 1906–1909 (1996).

  18. 18.

    , , , & Prevention of autoimmune symptoms in autoimmune-prone mice by elimination of B-1 cells. Int. Immunol. 7, 877–882 (1995).

  19. 19.

    , , & Ability of the xid gene to prevent autoimmunity in (NZB × NZW)F1 mice during the course of their natural history, after polyclonal stimulation, or following immunization with DNA. J. Clin. Invest. 70, 587–597 (1982).

  20. 20.

    , & Siglecs and their roles in the immune system. Nat. Rev. Immunol. 7, 255–266 (2007).

  21. 21.

    , , & Large-scale sequencing of the CD33-related Siglec gene cluster in five mammalian species reveals rapid evolution by multiple mechanisms. Proc. Natl. Acad. Sci. USA 101, 13251–13256 (2004).

  22. 22.

    Molecular diversity and evolution of the Siglec family of cell-surface lectins. Mol. Divers. 10, 555–566 (2006).

  23. 23.

    , , & Efficient targeting of the IL-4 gene in a BALB/c embryonic stem cell line. Transgenic Res. 5, 487–491 (1996).

  24. 24.

    & B1 cells: similarities and differences with other B cell subsets. Curr. Opin. Immunol. 13, 195–201 (2001).

  25. 25.

    , & The antigen recognized by MOMA-I is sialoadhesin. Immunol. Lett. 106, 96–98 (2006).

  26. 26.

    , , & Cloning and characterization of a novel mouse Siglec, mSiglec-F: differential evolution of the mouse and human (CD33) Siglec-3-related gene clusters. J. Biol. Chem. 276, 45128–45136 (2001).

  27. 27.

    et al. Identification, characterization and leucocyte expression of Siglec-10, a novel human sialic acid-binding receptor. Biochem. J. 355, 489–497 (2001).

  28. 28.

    et al. A new siglec family member, siglec-10, is expressed in cells of the immune system and has signaling properties similar to CD33. Eur. J. Biochem. 268, 6083–6096 (2001).

  29. 29.

    et al. Cloning and characterization of Siglec-10, a novel sialic acid binding member of the Ig superfamily, from human dendritic cells. J. Biol. Chem. 276, 28106–28112 (2001).

  30. 30.

    et al. A gene atlas of the mouse and human protein-encoding transcriptomes. Proc. Natl. Acad. Sci. USA 101, 6062–6067 (2004).

  31. 31.

    , , & Generation of long-lived B cells in germ-free mice. Eur. J. Immunol. 21, 1779–1782 (1991).

  32. 32.

    , & Bcmd decreases the life span of B-2 but not B-1 cells in A/WySnJ mice. J. Immunol. 160, 3743–3747 (1998).

  33. 33.

    , , & B-1a B cells that link the innate and adaptive immune responses are lacking in the absence of the spleen. J. Exp. Med. 195, 771–780 (2002).

  34. 34.

    et al. B cell-specific SHP1 deletion promotes B1a cell development and causes systemic autoimmunity. Immunity (in the press).

  35. 35.

    et al. Differentially regulated expression and function of CD22 in activated B- 1 and B-2 lymphocytes. J. Immunol. 168, 6078–6083 (2002).

  36. 36.

    , , , & The ligand-binding domain of CD22 is needed for inhibition of the B cell receptor signal, as demonstrated by a novel human CD22-specific inhibitor compound. J. Exp. Med. 195, 1207–1213 (2002).

  37. 37.

    , , & Sialic acid binding domains of CD22 are required for negative regulation of B cell receptor signaling. J. Exp. Med. 195, 1199–1205 (2002).

  38. 38.

    , , & Ablation of CD22 in ligand-deficient mice restores B cell receptor signaling. Nat. Immunol. 7, 199–206 (2006).

  39. 39.

    , & Regulation of B cell development and B cell signalling by CD22 and its ligands alpha2,6-linked sialic acids. Int. Immunol. 18, 603–611 (2006).

  40. 40.

    The role of CD22 and other inhibitory co-receptors in B-cell activation. Curr. Opin. Immunol. 17, 290–297 (2005).

  41. 41.

    et al. CD22 attenuates calcium signaling by potentiating plasma membrane calcium-ATPase activity. Nat. Immunol. 5, 651–657 (2004).

  42. 42.

    & Protein tyrosine phosphatase 1C negatively regulates antigen receptor signaling in B lymphocytes and determines thresholds for negative selection. Immunity 2, 13–24 (1995).

  43. 43.

    , & Marginal zone and B1 B cells unite in the early response against T-independent blood-borne particulate antigens. Immunity 14, 617–629 (2001).

  44. 44.

    et al. Reduction of marginal zone B cells in CD22-deficient mice. Eur. J. Immunol. 32, 561–567 (2002).

  45. 45.

    et al. The role of helper T cell products in mouse B cell differentiation and isotype regulation. Immunol. Rev. 102, 5–28 (1988).

  46. 46.

    et al. B cell defects in SLP65/BLNK-deficient mice can be partially corrected by the absence of CD22, an inhibitory coreceptor for BCR signaling. Eur. J. Immunol. 33, 3418–3426 (2003).

  47. 47.

    et al. Monoclonal antibodies to mouse complement receptor type 1 (CR1). Their use in a distribution study showing that mouse erythrocytes and platelets are CR1-negative. J. Immunol. 140, 3066–3072 (1988).

  48. 48.

    et al. Monoclonal antibodies specific for murine IgM I. Characterization of antigenic determinants on the four constant domains of the mu heavy chain. Eur. J. Immunol. 14, 534–542 (1984).

  49. 49.

    , & Altered selection processes of B lymphocytes in autoimmune NZB/W mice, despite intact central tolerance against DNA. Eur. J. Immunol. 31, 2800–2810 (2001).

Download references

Acknowledgements

We thank M. Döhler for blastocyst injections; C. Linden and U. Appelt for cell sorting; K. Voss, S. Angermüller and C. Dix for technical help; B. Bochner for Siglec-G cDNA; L. Klein for CD45.1 BALB/c mice; and R. Slany for help with retroviral infection. Supported by the Deutsche Forschungsgemeinschaft (SFB643 and FOR832).

Author information

Affiliations

  1. Department of Genetics, University of Erlangen, 91058 Erlangen, Germany.

    • Anja Hoffmann
    • , Julia Jellusova
    •  & Lars Nitschke
  2. Wellcome Trust Biocentre, University of Dundee, Dundee DD1 5EH, UK.

    • Sheena Kerr
    • , Jiquan Zhang
    •  & Paul R Crocker
  3. Department of Genetics, Nikolaus-Fiebiger-Zentrum, University of Erlangen, 91054 Erlangen, Germany.

    • Florian Weisel
    • , Ute Wellmann
    •  & Thomas H Winkler
  4. Zentrum für Operative Medizin, University of Würzburg, 97080 Würzburg, Germany.

    • Burkhard Kneitz

Authors

  1. Search for Anja Hoffmann in:

  2. Search for Sheena Kerr in:

  3. Search for Julia Jellusova in:

  4. Search for Jiquan Zhang in:

  5. Search for Florian Weisel in:

  6. Search for Ute Wellmann in:

  7. Search for Thomas H Winkler in:

  8. Search for Burkhard Kneitz in:

  9. Search for Paul R Crocker in:

  10. Search for Lars Nitschke in:

Contributions

A.H. did experiments and contributed to the writing; S.K., J.J., J.Z., F.W. and U.W. did experiments; T.H.W., B.K. and P.R.C. supervised experiments; and L.N. supervised experiments and wrote the paper.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Lars Nitschke.

Supplementary information

PDF files

  1. 1.

    Supplementary Fig. 1

    Alignment of murine Siglec-G and human Siglec-10.

  2. 2.

    Supplementary Fig. 2

    Siglec-G targeting strategy and resulting SiglecG-deficient mice.

  3. 3.

    Supplementary Fig. 3

    Proximal BCR-induced signaling is weaker in B1 cells than in B2 cells, but not substantially altered in Siglecg−/− mice.

  4. 4.

    Supplementary Fig. 4

    Autoantibodies in Siglecg−/− mice.

About this article

Publication history

Received

Accepted

Published

DOI

https://doi.org/10.1038/ni1480

Further reading