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

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Abstract

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.

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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.

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Acknowledgements

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

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.

Correspondence to Arthur Weiss.

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The authors declare no competing financial interests.

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