, Volume 61, Issue 12, pp 2621–2632 | Cite as

Silencing of high-affinity insulin-reactive B lymphocytes by anergy and impact of the NOD genetic background in mice

  • Mia J. Smith
  • Rochelle M. Hinman
  • Andrew Getahun
  • Soojin Kim
  • Thomas A. Packard
  • John C. CambierEmail author



Previous studies have demonstrated that high-affinity insulin-binding B cells (IBCs) silenced by anergy in healthy humans lose their anergy in islet autoantibody-positive individuals with recent-onset type 1 diabetes, and in autoantibody-negative first-degree relatives carrying certain risk alleles. Here we explore the hypothesis that IBCs are found in the immune periphery of disease-resistant C57BL/6-H2g7 mice, where, as in healthy humans, they are anergic, but that in disease-prone genetic backgrounds (NOD) they become activated and migrate to the pancreas and pancreatic lymph nodes, where they participate in the development of type 1 diabetes.


We compared the status of high-affinity IBCs in disease-resistant VH125.C57BL/6-H2g7 and disease-prone VH125.NOD mice.


Consistent with findings in healthy humans, high-affinity IBCs reach the periphery in disease-resistant mice and are anergic, as indicated by a reduced expression of membrane IgM, unresponsiveness to antigen and failure to become activated or accumulate in the pancreatic lymph nodes or pancreas. In NOD mice, high-affinity IBCs reach the periphery early in life and increase in number prior to the onset of hyperglycaemia. These cells are not anergic; they become activated, produce autoantibodies and accumulate in the pancreas and pancreatic lymph nodes prior to disease development.


These findings are consistent with genetic determination of the escape of high-affinity IBCs from anergy and their early contribution to the development of type 1 diabetes.


Anergy Autoantibodies Autoimmunity B cell Diabetes Insulin NOD 



B cell receptor


Insulin-binding B cell


Magnetic-activated cell sorting


Mean fluorescence intensity


Membrane IgM


Pancreatic lymph node


Phosphatase and tensin homologue


Spleen tyrosine kinase



The authors would like to thank J. W. Thomas (Vanderbilt University, Nashville, TN, USA) for providing the VH125.NOD and VH125.C57BL/6 mice.

Contribution statement

JCC designed the research and provided funding. MJS, RMH, TAP, AG, SK and JCC provided substantial contributions to conception and design, acquisition of data, and analysis and interpretation, as well as drafting the article and revising it critically; all authors have approved the version to be published. JCC is the guarantor of this work and, as such, had full access to all the data in the study and takes responsibility for the integrity of the data and the accuracy of the data analysis.


This work was supported by grants from the National Institutes of Health (DP3DK110845, R21AI124488, R01AI124487, R01DK096492 and F30OD021477).

Duality of interest

The authors declare that there is no duality of interest associated with this manuscript.


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Mia J. Smith
    • 1
    • 2
  • Rochelle M. Hinman
    • 1
  • Andrew Getahun
    • 1
  • Soojin Kim
    • 1
  • Thomas A. Packard
    • 1
  • John C. Cambier
    • 1
    Email author
  1. 1.Department of Immunology and MicrobiologyUniversity of Colorado School of MedicineAuroraUSA
  2. 2.Department of Microbiology, Immunology, and PathologyColorado State UniversityFort CollinsUSA

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