Perinatal oral exposure to low doses of bisphenol A, S or F impairs immune functions at intestinal and systemic levels in female offspring mice

Abstract

Background: Bisphenol A (BPA), one of the highest-volume chemicals produced worldwide, has been identified as an endocrine disruptor. Many peer-reviewing studies have reported adverse effects of low dose BPA exposure, particularly during perinatal period (gestation and/or lactation). We previously demonstrated that perinatal oral exposure to BPA (via gavage of mothers during gestation and lactation) has long-term consequences on immune response and intestinal barrier functions. Due to its adverse effects on several developmental and physiological processes, BPA was removed from consumer products and replaced by chemical substitutes such as BPS or BPF, that are structurally similar and not well studied compare to BPA. Here, we aimed to compare perinatal oral exposure to these bisphenols (BPs) at two doses (5 and 50 μg/kg of body weight (BW)/day (d)) on immune response at intestinal and systemic levels in female offspring mice at adulthood (Post Natal Day PND70).

Methods: Pregnant female mice were orally exposed to BPA, BPS or BPF at 5 or 50 μg/kg BW/d from 15th day of gravidity to weaning of pups at Post-Natal Day (PND) 21. Humoral and cellular immune responses of adult offspring (PND70) were analysed at intestinal and systemic levels.

Results: In female offspring, perinatal oral BP exposure led to adverse effects on intestinal and systemic immune response that were dependant of the BP nature (A, S or F) and dose of exposure. Stronger impacts were observed with BPS at the dose of 5 μg/kg BW/d on inflammatory markers in feces associated with an increase of anti-E. coli IgG in plasma. BPA and BPF exposure induced prominent changes at low dose in offspring mice, in term of intestinal and systemic immune responses, provoking an intestinal and systemic Th1/Th17 inflammation.

Conclusion: These findings provide, for the first time, results of long-time consequences of BPA, S and F perinatal exposure by oral route on immune response in offspring mice. This work warns that it is mandatory to consider immune markers and dose exposure in risk assessment associated to new BPA's alternatives.

Keywords: Bisphenol A; Bisphenol F; Bisphenol S; Cytokines; Immune responses; Immunoglobulin; Intestine; Perinatal exposure; Th1/Th17.

Fig. 1

Body weight of young and adult offspring mice. a Body weight of female offspring mice in different treatment groups measured at PND10 and b at PND70. * P < 0.05 vs. vehicle group. n = 5–14

Fig. 2

Perinatal exposure to bisphenols disrupts immune response in feces of adult offspring mice. a Total IgA concentration measured by ELISA in fecal samples of female offspring mice at PND70. b Lipocalin level determined in fecal supernatant of female offspring mice at PND70. * P < 0.05 vs. vehicle group. n = 5–14

Fig. 3

Perinatal exposure to bisphenols disrupts intestinal and systemic humoral responses in adult offspring dependent of bisphenol’s molecule. Plasma IgG (a) and IgA (b) concentrations measured by ELISA in female offspring mice at PND70. IgG (c) and IgA (d) specificity against E. coli lysate assessed in plasma by ELISA after normalizing to Ig concentrations. The lines represent the median (the 50th percentile). * P < 0.05; ** P < 0.01; **** P < 0.0001 vs. vehicle group. n = 5–14

Fig. 4

Perinatal exposure to BPA provokes intestinal Th1/Th17 immune response in adult offspring mice. Flow cytometry analysis of Th1 CD3⁺IFN-γ⁺T-bet⁺ (a) or Th17 CD3⁺RORγt⁺IL-17 ⁺ (c) lymphocytes from siLP. IFN-γ (b) or IL-17 (d) level assessed by ELISA after anti-CD3/CD28 in vitro restimulation of isolated lymphocytes from siLP offspring mice at PND70. (e) Proportion of CD4⁺CD25⁺FoxP3⁺ Treg cells in siLP of offspring mice at PND70. * P < 0.05 vs. vehicle group. n = 5–14

Fig. 5

Perinatal exposure to BPA or BPF provokes systemic Th1/Th17 immune response in adult offspring mice. Flow cytometry analysis of Th1 CD3⁺IFN-γ⁺T-bet⁺ (a) or Th17 CD3⁺RORγt⁺IL-17⁺ (c) lymphocytes from spleen. IFN-γ (b) or IL-17 (d) level assessed by ELISA after anti-CD3/CD28 in vitro restimulation of isolated lymphocytes from splenic offspring mice at PND70. (e) Proportion of CD4⁺CD25⁺FoxP3⁺ Treg cells in spleen of offspring mice at PND70. * P < 0.05; ** P < 0.01; *** P < 0.001 vs. vehicle group. n = 5–14

Fig. 6

Multivariate analysis representing immune profiles in function of BP perinatal exposure in offspring mice. a Sample score plot and associated loading plot on the first two PCA components resulting from all data set in offspring mice. Each color indicated groups with 0.85% confidence level ellipse plots. b PLS-DA sample score plot and associated loading plot (c) on the first two components derived from data set from all treated groups in offspring mice. Only loadings with correlation threshold > 0.5 were represented on the loading plots. % expl var.: percentages for each first two components explained by the model. n = 5–14