Sex-dependent effects of bisphenol A on type 1 diabetes development in non-obese diabetic (NOD) mice

  • Joella Xu
  • Guannan Huang
  • Tamas Nagy
  • Quincy Teng
  • Tai L. GuoEmail author
Organ Toxicity and Mechanisms


Type 1 diabetes (T1D) is an autoimmune disease caused by immune-mediated pancreatic β-cell destruction. The endocrine disrupting chemical bisphenol A (BPA) has widespread human exposure and can modulate immune function and the gut microbiome (GMB), which may contribute to the increasing T1D incidence worldwide. It was hypothesized that BPA had sex-dependent effects on T1D by modulating immune homeostasis and GMB. Adult female and male non-obese diabetic (NOD) mice were orally administered BPA at environmentally relevant doses (30 or 300 µg/kg). Antibiotic-treated adult NOD females were exposed to 0 or 30 µg/kg BPA. BPA accelerated T1D development in females, but delayed males from T1D. Consistently, females had a shift towards pro-inflammation (e.g., increased macrophages and Bacteroidetes), while males had increases in anti-inflammatory immune factors and a decrease in both anti- and pro-inflammatory GMB. Although bacteria altered during sub-acute BPA exposure differed from bacteria altered from chronic BPA exposure in both sexes, the GMB profile was consistently pro-inflammatory in females, while males had a general decrease of both anti- and pro-inflammatory gut microbes. However, treatment of females with the antibiotic vancomycin failed to prevent BPA-induced glucose intolerance, suggesting changes in Gram-positive bacteria were not a primary mechanism. In conclusion, BPA exposure was found to have sex dimorphic effects on T1D with detrimental effects in females, and immunomodulation was identified as the primary mechanism.


Bisphenol A Type 1 diabetes NOD mice Immunomodulation Microbiome Vancomycin 



The authors would like to thank Daniel E. Lefever, Dr. Travis Glenn and his lab members, and the Georgia Genomics and Bioinformatics Core of UGA for their help with the 16S rRNA library preparation, sequencing and bioinformatics analysis, and CVM Cytometry Core Facility (the College of Veterinary Medicine, UGA) for assisting flow cytometric analysis. This study was supported by NIH R21ES24487, and in part by NIH R41AT009523 and Interdisciplinary Toxicology Program at University of Georgia (UGA).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.

Supplementary material

204_2018_2379_MOESM1_ESM.pdf (992 kb)
Supplementary material 1 (PDF 991 KB)


  1. Abo T, Tomiyama C, Watanabe H (2012) Biology of autoreactive extrathymic T cells and B-1 cells of the innate immune system. Immunol Res 52:224–230CrossRefGoogle Scholar
  2. Alnek K, Kisand K, Heilman K, Peet A, Varik K, Uibo R (2015) Increased blood levels of growth factors, proinflammatory cytokines, and Th17 cytokines in patients with newly diagnosed type 1 diabetes PloS One 10:e0142976CrossRefGoogle Scholar
  3. Barthson J et al (2011) Cytokines tumor necrosis factor-α and interferon-γ induce pancreatic β-cell apoptosis through STAT1-mediated Bim protein activation. J Biol Chem 286:39632–39643CrossRefGoogle Scholar
  4. Belkaid Y, Hand TW (2014) Role of the microbiota in immunity and inflammation Cell 157:121–141CrossRefGoogle Scholar
  5. Bergman A et al (2013) The impact of endocrine disruption: a consensus statement on the state of the science. Environ Health Perspect 121:A104–A106. CrossRefPubMedPubMedCentralGoogle Scholar
  6. Beydoun HA, Khanal S, Zonderman AB, Beydoun MA (2014) Sex differences in the association of urinary bisphenol-A concentration with selected indices of glucose homeostasis among US adults. Ann Epidemiol 24:90–97CrossRefGoogle Scholar
  7. Bisikirska B, Colgan J, Luban J, Bluestone JA, Herold KC (2005) TCR stimulation with modified anti-CD3 mAb expands CD8 + T cell population and induces CD8 + CD25 + Tregs. J Clin Investig 115:2904–2913CrossRefGoogle Scholar
  8. Bodin J, Bolling AK, Samuelsen M, Becher R, Lovik M, Nygaard UC (2013) Long-term bisphenol A exposure accelerates insulitis development in diabetes-prone NOD mice Immunopharmacol Immunotoxicol 35:349–358. CrossRefPubMedGoogle Scholar
  9. Bodin J, Bolling AK, Becher R, Kuper F, Lovik M, Nygaard UC (2014) Transmaternal bisphenol A exposure accelerates diabetes type 1 development in NOD mice. Toxicol Sci 137:311–323. CrossRefPubMedGoogle Scholar
  10. Brown CT et al (2011) Gut microbiome metagenomics analysis suggests a functional model for the development of autoimmunity for type 1 diabetes PloS One 6:e25792CrossRefGoogle Scholar
  11. Calafat AM, Kuklenyik Z, Reidy JA, Caudill SP, Ekong J, Needham LL (2005) Urinary concentrations of bisphenol A and 4-nonylphenol in a human reference population. Environ Health Perspect 113:391–395CrossRefGoogle Scholar
  12. Cardozo AK, Proost P, Gysemans C, Chen MC, Mathieu C, Eizirik DL (2003) IL-1beta and IFN-gamma induce the expression of diverse chemokines and IL-15 in human and rat pancreatic islet cells, and in islets from pre-diabetic NOD mice Diabetologia 46:255–266. CrossRefPubMedGoogle Scholar
  13. Cetkovic-Cvrlje M, Thinamany S, Bruner KA (2017) Bisphenol A (BPA) aggravates multiple low-dose streptozotocin-induced type 1 diabetes in C57BL/6 mice. J Immunotoxicol 14:160–168CrossRefGoogle Scholar
  14. Chiang JL, Kirkman MS, Laffel LM, Peters AL (2014) Type 1 diabetes through the life span: a position statement of the American Diabetes Association Diabetes Care 37:2034–2054CrossRefGoogle Scholar
  15. Codella R et al (2015) Moderate intensity training impact on the inflammatory status and glycemic profiles in NOD mice. J Diabetes Res 2015:737586. CrossRefPubMedPubMedCentralGoogle Scholar
  16. Collins AM (2016) IgG subclass co-expression brings harmony to the quartet model of murine IgG function. Immunol Cell Biol 94:949–954CrossRefGoogle Scholar
  17. Cui X-B, Luan J-N, Ye J, Chen S-Y (2015) RGC32 deficiency protects against high-fat diet-induced obesity and insulin resistance in mice. J Endocrinol 224:127–137CrossRefGoogle Scholar
  18. de Goffau MC et al (2014) Aberrant gut microbiota composition at the onset of type 1 diabetes in young children. Diabetologia 57:1569–1577CrossRefGoogle Scholar
  19. Devaraj S, Tobias P, Jialal I (2011) Knockout of toll-like receptor-4 attenuates the pro-inflammatory state of diabetes Cytokine 55:441–445CrossRefGoogle Scholar
  20. Fu Z, Gilbert R, Liu E D (2013) Regulation of insulin synthesis and secretion and pancreatic beta-cell dysfunction in diabetes. Curr Diabetes Rev 9:25–53CrossRefGoogle Scholar
  21. Glenn TC et al (2016) Adapterama I: universal stubs and primers for thousands of dual-indexed Illumina libraries (iTru & iNext). BioRxiv. CrossRefGoogle Scholar
  22. Guo TL, Wang Y, Xiong T, Ling X, Zheng J (2014) Genistein modulation of streptozotocin diabetes in male B6C3F1 mice can be induced by diet. Toxicol Appl Pharmacol 280:455–466. CrossRefPubMedPubMedCentralGoogle Scholar
  23. Guo Y, Liu CQ, Shan CX, Chen Y, Li HH, Huang ZP, Zou DJ (2016) Gut microbiota after Roux-en-Y gastric bypass and sleeve gastrectomy in a diabetic rat model: increased diversity and associations of discriminant genera with metabolic changes Diabetes Metab Res Rev 33:e2857CrossRefGoogle Scholar
  24. Hansen CHF et al (2012) Early life treatment with vancomycin propagates Akkermansia muciniphila and reduces diabetes incidence in the NOD mouse Diabetologia 55:2285–2294CrossRefGoogle Scholar
  25. Hansen CH et al (2014) A maternal gluten-free diet reduces inflammation and diabetes incidence in the offspring of NOD mice. Diabetes 63:DB_131612CrossRefGoogle Scholar
  26. Huang G, Xu J, Lefever DE, Glenn TC, Nagy T, Guo TL (2017) Genistein prevention of hyperglycemia and improvement of glucose tolerance in adult non-obese diabetic mice are associated with alterations of gut microbiome and immune homeostasis Toxicol Appl Pharmacol CrossRefPubMedPubMedCentralGoogle Scholar
  27. Huxley RR, Peters SA, Mishra GD, Woodward M (2015) Risk of all-cause mortality and vascular events in women versus men with type 1 diabetes: a systematic review and meta-analysis. Lancet Diabetes Endocrinol 3:198–206. CrossRefPubMedGoogle Scholar
  28. Javurek AB, Spollen WG, Johnson SA, Bivens NJ, Bromert KH, Givan SA, Rosenfeld CS (2016) Effects of exposure to bisphenol A and ethinyl estradiol on the gut microbiota of parents and their offspring in a rodent model. Gut Microbes 7:471–485CrossRefGoogle Scholar
  29. Johnson SA et al (2016) Effects of developmental exposure to bisphenol A on spatial navigational learning and memory in rats: a CLARITY-BPA study. Horm Behav 80:139–148CrossRefGoogle Scholar
  30. Jörns A et al (2014) Islet infiltration, cytokine expression and beta cell death in the NOD mouse, BB rat, Komeda rat, LEW. 1AR1-iddm rat and humans with type 1 diabetes Diabetologia 57:512–521CrossRefGoogle Scholar
  31. Kaplan C, Valdez JC, Chandrasekaran R, Eibel H, Mikecz K, Glant TT, Finnegan A (2001) Th1 and Th2 cytokines regulate proteoglycan-specific autoantibody isotypes and arthritis. Arthritis Res Ther 4:54CrossRefGoogle Scholar
  32. Knip M, Siljander H (2016) The role of the intestinal microbiota in type 1 diabetes mellitus. Nat Rev Endocrinol 12:154CrossRefGoogle Scholar
  33. Krych Ł, Nielsen DS, Hansen AK, Hansen CHF (2015) Gut microbial markers are associated with diabetes onset, regulatory imbalance, and IFN-γ level in NOD Mice Gut Microbes 6:101–109CrossRefGoogle Scholar
  34. Lai K-P, Chung Y-T, Li R, Wan H-T, Wong CK-C (2016) Bisphenol A alters gut microbiome. Comparative metagenomics analysis Environ Pollut 218:923–930CrossRefGoogle Scholar
  35. Lang IA, Galloway TS, Scarlett A, Henley WE, Depledge M, Wallace RB, Melzer D (2008) Association of urinary bisphenol A concentration with medical disorders and laboratory abnormalities in adults Jama 300:1303–1310CrossRefGoogle Scholar
  36. Leeth CM et al (2016) B-Lymphocytes expressing an Ig specificity recognizing the pancreatic β-cell autoantigen peripherin are potent contributors to type 1 diabetes development in. NOD mice Diabetes 65:1977–1987CrossRefGoogle Scholar
  37. Lefever DE, Xu J, Chen Y, Huang G, Tamas N, Guo TL (2016) TCDD modulation of gut microbiome correlated with liver and immune toxicity in streptozotocin (STZ)-induced hyperglycemic mice Toxicol Appl Pharmacol 304:48–58CrossRefGoogle Scholar
  38. Liu Y, Yao Y, Li H, Qiao F, Wu J, Du Z-y, Zhang M (2016) Influence of endogenous and exogenous estrogenic endocrine on intestinal microbiota in zebrafish PloS One 11:e0163895CrossRefGoogle Scholar
  39. Malaisé Y et al (2017) Gut dysbiosis and impairment of immune system homeostasis in perinatally-exposed mice to bisphenol A precede obese phenotype development. Sci Rep 7:14472CrossRefGoogle Scholar
  40. Markle JG et al (2013) Sex differences in the gut microbiome drive hormone-dependent regulation of autoimmunity Science 339:1084–1088CrossRefGoogle Scholar
  41. Martin RM, Brady JL, Lew AM (1998) The need for IgG2c specific antiserum when isotyping antibodies from C57BL/6 and NOD mice. J Immunol Methods 212:187–192CrossRefGoogle Scholar
  42. Nadal A et al (2017) Extranuclear-initiated estrogenic actions of endocrine disrupting chemicals: is there toxicology beyond paracelsus? J Steroid Biochem Mol Biol. CrossRefPubMedGoogle Scholar
  43. Pitkäniemi J, Onkamo P, Tuomilehto J, Arjas E (2004) Increasing incidence of type 1 diabetes—role for genes? BMC Genet 5:5CrossRefGoogle Scholar
  44. Rubin BS, Paranjpe M, DaFonte T, Schaeberle C, Soto AM, Obin M, Greenberg AS (2017) Perinatal BPA exposure alters body weight and composition in a dose specific and sex specific manner: the addition of peripubertal exposure exacerbates adverse effects in female mice. Reprod Toxicol 68:130–144CrossRefGoogle Scholar
  45. Ryba-Stanislawowska M, Werner P, Brandt A, Mysliwiec M, Mysliwska J (2016) Th9 and Th22 immune response in young patients with type 1 diabetes Immunol Res 64:730–735. CrossRefPubMedGoogle Scholar
  46. Scinicariello F, Buser MC (2016) Serum testosterone concentrations and urinary bisphenol A, benzophenone-3, triclosan, and paraben levels in male and female children and adolescents: NHANES 2011–2012. Environ Health Perspect 124:1898CrossRefGoogle Scholar
  47. Segata N, Izard J, Waldron L, Gevers D, Miropolsky L, Garrett WS, Huttenhower C (2011) Metagenomic biomarker discovery and explanation Genome Biol 12:R60 CrossRefPubMedPubMedCentralGoogle Scholar
  48. Susiarjo M, Xin F, Bansal A, Stefaniak M, Li C, Simmons RA, Bartolomei MS (2015) Bisphenol a exposure disrupts metabolic health across multiple generations in the mouse Endocrinology 156:2049–2058CrossRefGoogle Scholar
  49. Taylor JA et al (2011) Similarity of bisphenol A pharmacokinetics in rhesus monkeys and mice: relevance for human exposure. Environ Health Perspect 119:422CrossRefGoogle Scholar
  50. Todd I, Davenport C, Topping JH, Wood PJ (1998) IgG2a antibodies non-specifically delay the onset of diabetes in NOD mice Autoimmunity 27:209–211CrossRefGoogle Scholar
  51. Tuller T, Atar S, Ruppin E, Gurevich M, Achiron A (2013) Common and specific signatures of gene expression and protein–protein interactions in autoimmune diseases. Genes Immun 14:67–82CrossRefGoogle Scholar
  52. Xu J, Huang G, Guo TL (2016) Developmental bisphenol A exposure modulates. Immune Relat Dis Toxics 4:23Google Scholar
  53. Yoshino S, Yamaki K, Yanagisawa R, Takano H, Hayashi H, Mori Y (2003) Effects of bisphenol A on antigen-specific antibody production, proliferative responses of lymphoid cells, and TH1 and TH2 immune responses in mice. Br J Pharmacol 138:1271–1276CrossRefGoogle Scholar
  54. Young EF, Hess PR, Arnold LW, Tisch R, Frelinger JA (2009) Islet lymphocyte subsets in male and female NOD mice are qualitatively similar but quantitatively distinct Autoimmunity 42:678–691CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Veterinary Biosciences and Diagnostic Imaging, College of Veterinary MedicineUniversity of GeorgiaAthensUSA
  2. 2.Department of Environmental Health SciencesUniversity of GeorgiaAthensUSA
  3. 3.Department of PathologyUniversity of GeorgiaAthensUSA
  4. 4.Department of Pharmaceutical and Biomedical Sciences, College of PharmacyUniversity of GeorgiaAthensUSA

Personalised recommendations