pp 1–9 | Cite as

Regulation of B cell homeostasis by Ptpn22 contributes to type 1 diabetes in NOD mice

  • Xiajie Shi
  • Feng Shao
  • Zhixia Li
  • Lin Kang
  • Junbin Liu
  • Stephan Kissler
  • Zhiguang ZhouEmail author
  • Lijing JiaEmail author
  • Peilin ZhengEmail author
Original Article



A coding variant in PTPN22 (C1858T) is one of the most important genetic risk factors in type 1 diabetes (T1D). The role of the PTPN22 risk allele in B cells is still incompletely understood and has not been investigated directly in T1D. This study aimed to explore the role of PTPN22 in the homeostasis of B cells and its influence in T1D.


Wild-type (WT) and Ptpn22 inducible knockdown (KD) NOD mice were treated with 200 μg/ml doxycycline at the age of 10 weeks for 1–2 months. B cell compositions in the bone marrow, peritoneal cavity and spleen were examined. The pathogenicity of Ptpn22 KD B cells was explored by adoptive cell transfer.


Ptpn22 silencing increased the frequency of recirculating mature B cells in the bone marrow, decreased the frequency of B-1a cells in the peritoneal cavity and suppressed the formation of marginal zone B cells and plasma cells in the spleen. Changes in the composition of the peripheral B cell compartment caused by altered cell proliferation while rates of apoptosis were not affected. Significantly, co-transfer of Ptpn22 KD B cells with NY8.3 diabetogenic T cells diminished the frequency of diabetes in recipient NOD.scid mice compared with co-transfer of WT B cells.


Our study constitutes the first functional study of Ptpn22 in B cells in NOD mice. Our findings suggest that Ptpn22 variation contributes to T1D by modifying the B cell compartment and support a gain-of-function for the PTPN22 disease variant.


Type 1 diabetes PTPN22 B cells Autoimmunity 



Type 1 diabetes


Anti-insulin antibodies


Regulatory B cells


B cell receptor




Non obese diabetic


Wild type




Marginal zone




Plasmacytoid dendtric cells



This work is supported by the National Natural Science Foundation of China (Grant No. 81670716, 81500600), Hunan Natural Science Fund for Excellent Young Scholars (Grant No. 2019JJ30036), and the Graduate innovation project of Central South University (Grant No. 2018zzts921). The authors declare no conflicts of interest.

Author contributions

All authors have read and approved the final manuscript. P.Z., L.J., and Z.Z. discussed, designed the study and critically edited the manuscript. X.S. and F.S. conducted the experiments, analysed data, and wrote the manuscript. Z.L., L.K., and J.L. contributed to the experiments and discussion. S.K. provided P2 transgenic mice and critically edited the manuscript.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants performed by any of the authors. All applicable institutional guidelines for the care and use of animals were followed.

Supplementary material

12020_2019_2120_MOESM1_ESM.docx (3 mb)
Supplementary Information


  1. 1.
    J.A. Todd, Etiology of type 1 diabetes. Immunity 32, 457–467 (2010)CrossRefGoogle Scholar
  2. 2.
    L.A. DiMeglio, C. Evans-Molina, R.A. Oram, Type 1 diabetes. Lancet 391, 2449–2462 (2018)CrossRefGoogle Scholar
  3. 3.
    J. Diana, Y. Simoni, L. Furio, L. Beaudoin, B. Agerberth, F. Barrat, A. Lehuen, Crosstalk between neutrophils, B-1a cells and plasmacytoid dendritic cells initiates autoimmune diabetes. Nat. Med. 19, 65–73 (2012)CrossRefGoogle Scholar
  4. 4.
    R.M. Hinman, J.C. Cambier, Role of B lymphocytes in the pathogenesis of type 1 diabetes. Curr. Diab. Rep. 14(11), 543 (2014)CrossRefGoogle Scholar
  5. 5.
    J.P. Palmer, C.M. Asplin, P. Clemons, K. Lyen, O. Tatpati, P.K. Raghu, T.L. Paquette, Insulin antibodies in insulin-dependent diabetics before insulin treatment. Science 222, 1337–1339 (1983)CrossRefGoogle Scholar
  6. 6.
    G. De Filippo, N. Pozzi, E. Cosentini, M. Cavalcanti, J.C. Carel, S. Tamasi, A. Franzese, C. Pignata, Increased CD5+CD19+ B lymphocytes at the onset of type 1 diabetes in children. Acta Diabetol. 34, 271–274 (1997)CrossRefGoogle Scholar
  7. 7.
    C. Deng, Y. Xiang, T. Tan, Z. Ren, C. Cao, G. Huang, L. Wen, Z. Zhou, Altered peripheral B-lymphocyte subsets in type 1 diabetes and latent autoimmune diabetes in adults. Diabetes Care 39, 434–440 (2016)CrossRefGoogle Scholar
  8. 8.
    M.D. Pescovitz, C.J. Greenbaum, H. Krause-Steinrauf, D.J. Becker, S.E. Gitelman, R. Goland, P.A. Gottlieb, J.B. Marks, P.F. McGee, A.M. Moran, P. Raskin, H. Rodriguez, D.A. Schatz, D. Wherrett, D.M. Wilson, J.M. Lachin, J.S. Skyler, Type 1 diabetes Trialnet anti-CD20 Study Group, rituximab, B-lymphocyte depletion, and preservation of beta-cell function. N. Engl. J. Med. 361, 2143–2152 (2009)CrossRefGoogle Scholar
  9. 9.
    M.D. Pescovitz, C.J. Greenbaum, B. Bundy, D.J. Becker, S.E. Gitelman, R. Goland, P.A. Gottlieb, J.B. Marks, A. Moran, P. Raskin, H. Rodriguez, D.A. Schatz, D.K. Wherrett, D.M. Wilson, J.P. Krischer, J.S. Skyler, Type 1 diabetes TrialNet Anti-CD20 Study Group, B-lymphocyte depletion with rituximab and beta-cell function: two-year results. Diabetes Care 37, 453–459 (2014)CrossRefGoogle Scholar
  10. 10.
    D.V. Serreze, H.D. Chapman, D.S. Varnum, M.S. Hanson, P.C. Reifsnyder, S.D. Richard, S.A. Fleming, E.H. Leiter, L.D. Shultz, B lymphocytes are essential for the initiation of T cell-mediated autoimmune diabetes: analysis of a new “speed congenic” stock of NOD.Ig mu null mice. J. Exp. Med. 184, 2049–2053 (1996)CrossRefGoogle Scholar
  11. 11.
    N. Bottini, L. Musumeci, A. Alonso, S. Rahmouni, K. Nika, M. Rostamkhani, J. MacMurray, G.F. Meloni, P. Lucarelli, M. Pellecchia, G.S. Eisenbarth, D. Comings, T. Mustelin, A functional variant of lymphoid tyrosine phosphatase is associated with type I diabetes. Nat. Genet. 36, 337–338 (2004)CrossRefGoogle Scholar
  12. 12.
    T. Vang, M. Congia, M.D. Macis, L. Musumeci, V. Orrú, P. Zavattari, K. Nika, L. Tautz, K. Taskén, F. Cucca, T. Mustelin, N. Bottini, Autoimmune-associated lymphoid tyrosine phosphatase is a gain-of-function variant. Nat. Genet. 37, 1317–1319 (2005)CrossRefGoogle Scholar
  13. 13.
    X. Dai, R.G. James, T. Habib, S. Singh, S. Jackson, S. Khim, R.T. Moon, D. Liggitt, A. Wolf-Yadlin, J.H. Buckner, D.J. Rawlings, A disease-associated PTPN22 variant promotes systemic autoimmunity in murine models. J. Clin. Investig. 123, 2024–2036 (2013)CrossRefGoogle Scholar
  14. 14.
    L. Menard, D. Saadoun, I. Isnardi, Y.S. Ng, G. Meyers, C. Massad, C. Price, C. Abraham, R. Motaghedi, J.H. Buckner, P.K. Gregersen, E. Meffre, The PTPN22 allele encoding an R620W variant interferes with the removal of developing autoreactive B cells in humans. J. Clin. Investig. 121, 3635–3644 (2011)CrossRefGoogle Scholar
  15. 15.
    X. Lin, S. Pelletier, S. Gingras, S. Rigaud, C.J. Maine, K. Marquardt, Y.D. Dai, K. Sauer, A.R. Rodriguez, G. Martin, S. Kupriyanov, L. Jiang, L. Yu, D.R. Green, L.A. Sherman, CRISPR-Cas9-mediated modification of the NOD mouse genome with Ptpn22R619W mutation increases autoimmune diabetes. Diabetes 65, 2134–2138 (2016)CrossRefGoogle Scholar
  16. 16.
    T. Habib, A. Funk, M. Rieck, A. Brahmandam, X. Dai, A.K. Panigrahi, E.T. Luning Prak, A. Meyer-Bahlburg, S. Sanda, C. Greenbaum, D.J. Rawlings, J.H. Buckner, Altered B cell homeostasis is associated with type I diabetes and carriers of the PTPN22 allelic variant. J. Immunol. 188, 487–496 (2012)CrossRefGoogle Scholar
  17. 17.
    G. Metzler, X. Dai, C.D. Thouvenel, S. Khim, T. Habib, J.H. Buckner, D.J. Rawlings, The autoimmune risk variant PTPN22 C1858T alters B cell tolerance at discrete checkpoints and differentially shapes the naive repertoire. J. Immunol. 199, 2249–2260 (2017)CrossRefGoogle Scholar
  18. 18.
    P. Zheng, S. Kissler, PTPN22 silencing in the NOD model indicates the type 1 diabetes-associated allele is not a loss-of-function variant. Diabetes 62, 896–904 (2013)CrossRefGoogle Scholar
  19. 19.
    C. Schuster, K.D. Gerold, K. Schober, L. Probst, K. Boerner, M.J. Kim, A. Ruckdeschel, T. Serwold, S. Kissler, The autoimmunity-associated gene CLEC16A modulates thymic epithelial cell autophagy and alters T cell selection. Immunity 42, 942–952 (2015)CrossRefGoogle Scholar
  20. 20.
    P. Schneider, H. Takatsuka, A. Wilson, F. Mackay, A. Tardivel, S. Lens, T.G. Cachero, D. Finke, F. Beermann, J. Tschopp, Maturation of marginal zone and follicular B cells requires B cell activating factor of the tumor necrosis factor family and is independent of B cell maturation antigen. J. Exp. Med. 194, 1691–1697 (2001)CrossRefGoogle Scholar
  21. 21.
    J.N. Schickel, M. Kuhny, A. Baldo, J.M. Bannock, C. Massad, H. Wang, N. Katz, T. Oe, L. Menard, P. Soulas-Sprauel, T. Strowig, R. Flavell, E. Meffre, PTPN22 inhibition resets defective human central B cell tolerance. Sci. Immunol. 1, f7153 (2016)CrossRefGoogle Scholar
  22. 22.
    S.B. Hartley, J. Crosbie, R. Brink, A.B. Kantor, A. Basten, C.C. Goodnow, Elimination from peripheral lymphoid tissues of self-reactive B lymphocytes recognizing membrane-bound antigens. Nature 353, 765–769 (1991)CrossRefGoogle Scholar
  23. 23.
    M.S. Anderson, J.A. Bluestone, The NOD mouse: a model of immune dysregulation. Annu. Rev. Immunol. 23, 447–485 (2005)CrossRefGoogle Scholar
  24. 24.
    G.J. Tsay, M. Zouali, The interplay between innate-like B cells and other cell types in autoimmunity. Front Immunol. 9, 1064 (2018)CrossRefGoogle Scholar
  25. 25.
    J.J. Kenny, A.M. Stall, D.G. Sieckmann et al. Receptor-mediated elimination of phosphocholine-specific B cells in x-linked immune-deficient mice. J. Immunol. 146, 2568–2577 (1991)PubMedGoogle Scholar
  26. 26.
    L. Mandik-Nayak, J. Racz, B.P. Sleckman, M.C. Lamers, F.D. Finkelman, L. Finch, D.L. Longo, Autoreactive marginal zone B cells are spontaneously activated but lymph node B cells require T cell help. J. Exp. Med. 203, 1985–1998 (2006)CrossRefGoogle Scholar
  27. 27.
    K. Attanavanich, J.F. Kearney, Marginal zone, but not follicular B cells, are potent activators of naive CD4 T cells. J. Immunol. 172, 803–811 (2004)CrossRefGoogle Scholar
  28. 28.
    E. Marino, M. Batten, J. Groom, S. Walters, D. Liuwantara, F. Mackay, S.T. Grey, Marginal-zone B-cells of nonobese diabetic mice expand with diabetes onset, invade the pancreatic lymph nodes, and present autoantigen to diabetogenic T-cells. Diabetes 57, 395–404 (2008)CrossRefGoogle Scholar
  29. 29.
    E. Marino, J.L. Richards, K.H. McLeod, D. Stanley, Y.A. Yap, J. Knight, C. McKenzie, J. Kranich, A.C. Oliveira, F.J. Rossello, B. Krishnamurthy, C.M. Nefzger, L. Macia, A. Thorburn, A.G. Baxter, G. Morahan, L.H. Wong, J.M. Polo, R.J. Moore, T.J. Lockett, J.M. Clarke, D.L. Topping, L.C. Harrison, C.R. Mackay, Gut microbial metabolites limit the frequency of autoimmune T cells and protect against type 1 diabetes. Nat. Immunol. 18, 552–562 (2017)CrossRefGoogle Scholar
  30. 30.
    P. Zheng, Z. Li, Z. Zhou, Gut microbiome in type 1 diabetes: a comprehensive review. Diabetes Metabol. Res. Rev. 34, e3043 (2018)CrossRefGoogle Scholar
  31. 31.
    L.I. Pao, K.P. Lam, J.M. Henderson, J.L. Kutok, M. Alimzhanov, L. Nitschke, M.L. Thomas, B.G. Neel, K. Rajewsky, B cell-specific deletion of protein-tyrosine phosphatase Shp1 promotes B-1a cell development and causes systemic autoimmunity. Immunity 27, 35–48 (2007)CrossRefGoogle Scholar
  32. 32.
    J. Corte-Real, N. Duarte, L. Tavares, C. Penha-Gonçalves, Innate stimulation of B1a cells enhances the autoreactive IgM repertoire in the NOD mouse: implications for type 1 diabetes. Diabetologia 55, 1761–1772 (2012)CrossRefGoogle Scholar
  33. 33.
    P.L. Kendall, E.J. Woodward, C. Hulbert, J.W. Thomas, Peritoneal B cells govern the outcome of diabetes in non-obese diabetic mice. Eur. J. Immunol. 34, 2387–2395 (2004)CrossRefGoogle Scholar
  34. 34.
    R.A. Smerdon, M. Peakman, M.J. Hussain et al. CD5+ B-cells at the onset of type I diabetes and in the prediabetic period. Diabetes Care 17, 657–664 (1994)CrossRefGoogle Scholar
  35. 35.
    C. Hulbert, B. Riseili, M. Rojas, J.W. Thomas, B cell specificity contributes to the outcome of diabetes in nonobese diabetic mice. J. Immunol. 167, 5535–5538 (2001)CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Metabolism & Endocrinology, The Second Xiangya Hospital, Key Laboratory of Diabetes Immunology, Central South University, Ministry of EducationNational Clinical Research Center for Metabolic DiseasesChangshaChina
  2. 2.Department of Endocrinology, Shenzhen People’s Hospital, The Second Clinical Medical College of Jinan UniversityThe First Affiliated Hospital of Southern University of Science and TechnologyShenzhenChina
  3. 3.Section for Immunobiology, Joslin Diabetes CenterHarvard Medical SchoolBostonUSA

Personalised recommendations