Skip to main content

Thank you for visiting You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Endogenous antigen tunes the responsiveness of naive B cells but not T cells


In humans, up to 75% of newly generated B cells and about 30% of mature B cells show some degree of autoreactivity1. Yet, how B cells establish and maintain tolerance in the face of autoantigen exposure during and after development is not certain. Studies of model B-cell antigen receptor (BCR) transgenic systems have highlighted the critical role of functional unresponsiveness or ‘anergy’2,3. Unlike T cells, evidence suggests that receptor editing and anergy, rather than deletion, account for much of B-cell tolerance4,5. However, it remains unclear whether the mature diverse B-cell repertoire of mice contains anergic autoreactive B cells, and if so, whether antigen was encountered during or after their development. By taking advantage of a reporter mouse in which BCR signalling rapidly and robustly induces green fluorescent protein expression under the control of the Nur77 regulatory region, antigen-dependent and antigen-independent BCR signalling events in vivo during B-cell maturation were visualized. Here we show that B cells encounter antigen during development in the spleen, and that this antigen exposure, in turn, tunes the responsiveness of BCR signalling in B cells at least partly by downmodulating expression of surface IgM but not IgD BCRs, and by modifying basal calcium levels. By contrast, no analogous process occurs in naive mature T cells. Our data demonstrate not only that autoreactive B cells persist in the mature repertoire, but that functional unresponsiveness or anergy exists in the mature B-cell repertoire along a continuum, a fact that has long been suspected, but never yet shown. These results have important implications for understanding how tolerance in T and B cells is differently imposed, and how these processes might go awry in disease.

This is a preview of subscription content, access via your institution

Relevant articles

Open Access articles citing this article.

Access options

Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.

Figure 1: The Nur77–GFP BAC transgenic reporter is responsive to antigen-receptor signalling in vitro.
Figure 2: Expression of the Nur77–GFP BAC transgenic reporter is upregulated at specific checkpoints during B-cell development.
Figure 3: Expression of the Nur77–GFP BAC transgenic reporter is sensitive to genetic modulation of BCR signal strength and to antigen.
Figure 4: GFP expression predicts functional responsiveness and autoreactivity of B cells.


  1. Wardemann, H. et al. Predominant autoantibody production by early human B cell precursors. Science 301, 1374–1377 (2003)

    ADS  CAS  Article  Google Scholar 

  2. Goodnow, C. C. et al. Altered immunoglobulin expression and functional silencing of self-reactive B lymphocytes in transgenic mice. Nature 334, 676–682 (1988)

    ADS  CAS  Article  Google Scholar 

  3. Cambier, J. C., Gauld, S. B., Merrell, K. T. & Vilen, B. J. B-cell anergy: from transgenic models to naturally occurring anergic B cells? Nature Rev. Immunol. 7, 633–643 (2007)

    CAS  Article  Google Scholar 

  4. Lang, J. et al. Enforced Bcl-2 expression inhibits antigen-mediated clonal elimination of peripheral B cells in an antigen dose-dependent manner and promotes receptor editing in autoreactive, immature B cells. J. Exp. Med. 186, 1513–1522 (1997)

    CAS  Article  Google Scholar 

  5. Halverson, R., Torres, R. & Pelanda, R. Receptor editing is the main mechanism of B cell tolerance toward membrane antigens. Nature Immunol. 5, 645–650 (2004)

    CAS  Article  Google Scholar 

  6. Moran, A. E. et al. T cell receptor signal strength in Treg and iNKT cell development demonstrated by a novel fluorescent reporter mouse. J. Exp. Med. 208, 1279–1289 (2011)

    CAS  Article  Google Scholar 

  7. Winoto, A. & Littman, D. R. Nuclear hormone receptors in T lymphocytes. Cell 109 (Suppl. 1). S57–S66 (2002)

    CAS  Article  Google Scholar 

  8. Mittelstadt, P. R. & DeFranco, A. L. Induction of early response genes by cross-linking membrane Ig on B lymphocytes. J. Immunol. 150, 4822–4832 (1993)

    CAS  PubMed  Google Scholar 

  9. The Gene Expression Nervous System Atlas (GENSAT) Project. NINDS Contract # N01NS02331 to The Rockefeller University (New York, USA)

  10. Zikherman, J. et al. CD45–Csk phosphatase–kinase titration uncouples basal and inducible T cell receptor signaling during thymic development. Immunity 32, 342–354 (2010)

    CAS  Article  Google Scholar 

  11. Hardy, R. R., Carmack, C. E., Shinton, S. A., Kemp, J. D. & Hayakawa, K. Resolution and characterization of pro-B and pre-pro-B cell stages in normal mouse bone marrow. J. Exp. Med. 173, 1213–1225 (1991)

    CAS  Article  Google Scholar 

  12. Goodnow, C. C., Sprent, J., Fazekas de St Groth, B. & Vinuesa, C. G. Cellular and genetic mechanisms of self tolerance and autoimmunity. Nature 435, 590–597 (2005)

    ADS  CAS  Article  Google Scholar 

  13. Loder, F. et al. B cell development in the spleen takes place in discrete steps and is determined by the quality of B cell receptor-derived signals. J. Exp. Med. 190, 75–90 (1999)

    CAS  Article  Google Scholar 

  14. Chung, J. B., Silverman, M. & Monroe, J. G. Transitional B cells: step by step towards immune competence. Trends Immunol. 24, 342–349 (2003)

    Article  Google Scholar 

  15. Allman, D. et al. Resolution of three nonproliferative immature splenic B cell subsets reveals multiple selection points during peripheral B cell maturation. J. Immunol. 167, 6834–6840 (2001)

    CAS  Article  Google Scholar 

  16. Zikherman, J., Doan, K., Parameswaran, R., Raschke, W. & Weiss, A. Quantitative differences in CD45 expression unmask functions for CD45 in B-cell development, tolerance, and survival. Proc. Natl Acad. Sci. USA 109, E3–E12 (2012)

    ADS  CAS  Article  Google Scholar 

  17. Cooke, M. et al. Immunoglobulin signal transduction guides the specificity of B cell–T cell interactions and is blocked in tolerant self-reactive B cells. J. Exp. Med. 179, 425–438 (1994)

    CAS  Article  Google Scholar 

  18. Yarkoni, Y., Getahun, A. & Cambier, J. C. Molecular underpinning of B-cell anergy. Immunol. Rev. 237, 249–263 (2010)

    CAS  Article  Google Scholar 

  19. Duty, J. A. et al. Functional anergy in a subpopulation of naive B cells from healthy humans that express autoreactive immunoglobulin receptors. J. Exp. Med. 206, 139–151 (2009)

    Article  Google Scholar 

  20. Glynne, R. et al. How self-tolerance and the immunosuppressive drug FK506 prevent B-cell mitogenesis. Nature 403, 672–676 (2000)

    ADS  CAS  Article  Google Scholar 

  21. Glynne, R., Ghandour, G., Rayner, J., Mack, D. H. & Goodnow, C. C. B-lymphocyte quiescence, tolerance and activation as viewed by global gene expression profiling on microarrays. Immunol. Rev. 176, 216–246 (2000)

    CAS  Article  Google Scholar 

  22. Merrell, K. et al. Identification of anergic B cells within a wild-type repertoire. Immunity 25, 953–962 (2006)

    CAS  Article  Google Scholar 

  23. Teague, B. N. et al. Cutting edge: transitional T3 B cells do not give rise to mature B cells, have undergone selection, and are reduced in murine lupus. J. Immunol. 178, 7511–7515 (2007)

    CAS  Article  Google Scholar 

  24. Cornall, R. J. et al. Polygenic autoimmune traits: Lyn, CD22, and SHP-1 are limiting elements of a biochemical pathway regulating BCR signaling and selection. Immunity 8, 497–508 (1998)

    CAS  Article  Google Scholar 

  25. Benschop, R. J. et al. Activation and anergy in bone marrow B cells of a novel immunoglobulin transgenic mouse that is both hapten specific and autoreactive. Immunity 14, 33–43 (2001)

    CAS  Article  Google Scholar 

  26. Fields, M. L. & Erikson, J. The regulation of lupus-associated autoantibodies: immunoglobulin transgenic models. Curr. Opin. Immunol. 15, 709–717 (2003)

    CAS  Article  Google Scholar 

  27. Virts, E. L., Diago, O. & Raschke, W. C. A. CD45 minigene restores regulated isoform expression and immune function in CD45-deficient mice: therapeutic implications for human CD45-null severe combined immunodeficiency. Blood 101, 849–855 (2003)

    CAS  Article  Google Scholar 

  28. Zikherman, J. et al. PTPN22 deficiency cooperates with the CD45 E613R allele to break tolerance on a non-autoimmune background. J. Immunol. 182, 4093–4106 (2009)

    CAS  Article  Google Scholar 

  29. Hermiston, M. L., Tan, A. L., Gupta, V. A., Majeti, R. & Weiss, A. The juxtamembrane wedge negatively regulates CD45 function in B cells. Immunity 23, 635–647 (2005)

    CAS  Article  Google Scholar 

Download references


We thank A. Roque for assisting with animal husbandry and Z. Wang and J. Paw for help with cell sorting. This work was supported by the Rosalind Russell Medical Research Foundation Bechtel Award (J.Z.), an American College of Rheumatology REF Rheumatology Investigator Award (J.Z.), an Arthritis National Research Foundation grant (J.Z.) and National Institutes of Health Grant K08 AR059723 (J.Z.), as well as the Howard Hughes Medical Institute (A.W.).

Author information

Authors and Affiliations



J.Z. and A.W. designed the research, J.Z. and R.P. performed the research, J.Z. and R.P. analysed the data and J.Z. and A.W. wrote the manuscript.

Corresponding author

Correspondence to Arthur Weiss.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Information

This file contains Supplementary Figures 1-10. (PDF 3578 kb)

PowerPoint slides

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Zikherman, J., Parameswaran, R. & Weiss, A. Endogenous antigen tunes the responsiveness of naive B cells but not T cells. Nature 489, 160–164 (2012).

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI:

Further reading


By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.


Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing