Although it is generally accepted that dietary fiber is health promoting, the underlying immunological and molecular mechanisms are not well defined, especially with respect to cellulose, the most ubiquitous dietary fiber. Here, the impact of dietary cellulose on intestinal microbiota, immune responses and gene expression in health and disease was examined. Lack of dietary cellulose disrupted the age-related diversification of the intestinal microbiota, which subsequently remained in an immature state. Interestingly, one of the most affected microbial genera was Alistipes which is equipped with enzymes to degrade cellulose. Absence of cellulose changed the microbial metabolome, skewed intestinal immune responses toward inflammation, altered the gene expression of intestinal epithelial cells and mice showed increased sensitivity to colitis induction. In contrast, mice with a defined microbiota including A. finegoldii showed enhanced colonic expression of intestinal IL-22 and Reg3γ restoring intestinal barrier function. This study supports the epidemiological observations and adds a causal explanation for the health promoting effects of the most common biopolymer on earth.

Endocrine disrupting chemicals (EDCs) possess the capability to interfere with the endocrine system by binding to hormone receptors, for example on immune cells. Specific effects have already been described for individual substances, but the impact of exposure to chemical mixtures during pregnancy on maternal immune regulation, placentation and fetal development is not known. In this study, we aimed to investigate the combined effects of two widespread EDCs, bisphenol A (BPA) and benzophenone-3 (BP-3), at allowed concentrations on crucial pregnancy processes such as implantation, placentation, uterine immune cell populations and fetal growth. From gestation day (gd) 0 to gd10, female mice were exposed to 4 μg/kg/d BPA, 50 mg/kg/d BP-3 or a BPA/BP-3 mixture. High frequency ultrasound and Doppler measurements were used to determine intrauterine fetal development and hemodynamic parameters. Furthermore, uterine spiral artery remodeling and placental mRNA expression were studied via histology and CHIP-RT-PCR, respectively. Effects of EDC exposure on multiple uterine immune cell populations were investigated using flow cytometry. We found that exposure to BP-3 caused intrauterine growth restriction in offspring at gd14, while BPA and BPA/BP-3 mixture caused varying effects. Moreover, placental morphology at gd12 and placental efficiency at gd14 were altered upon BP-3 exposure. Placental gene transcription was altered particularly in female offspring after in utero exposure to BP-3. Flow cytometry analyses revealed an increase in uterine T cells and NK cells in BPA and BPA/BP-3-treated dams at gd14. Doppler measurements revealed no effect on uterine hemodynamic parameters and spiral artery remodeling was not affected following EDC exposure. Our results provide evidence that exposure to BPA and BP-3 during early gestation affects fetal development in a sex-dependent manner, placental function and immune cell frequencies at the feto-maternal interface. These results call for inclusion of studies addressing pregnancy in the risk assessment of environmental chemicals.

Immune homeostasis is key to guarantee that the immune system can elicit effector functions against pathogens and at the same time raise tolerance towards other antigens. A disturbance of this delicate balance may underlie or at least trigger pathologies. Endocrine disrupting chemicals (EDCs) are increasingly recognized as risk factors for immune dysregulation. However, the immunotoxic potential of specific EDCs and their mixtures is still poorly understood. Thus, we aimed to investigate the effect of bisphenol A (BPA) and benzophenone-3 (BP-3), alone and in combination, on in vitro differentiation of T helper (TH)17 cells and regulatory T (T_reg) cells. Naïve T cells were isolated from mouse lymphoid tissues and differentiated into the respective TH population in the presence of 0.001-10 μM BP-3 and/or 0.01-100 μM BPA. Cell viability, proliferation and the expression of TH lineage specific transcription factors and cytokines was measured by flow cytometry and CBA/ELISA. Moreover, the transcription of hormone receptors as direct targets of EDCs was quantified by RT-PCR. We found that the highest BPA concentration adversely affected TH cell viability and proliferation. Moreover, the general differentiation potential of both TH populations was not altered in the presence of both EDCs. However, EDC exposure modulated the emergence of TH17 and T_reg cell intermediate states. While BPA and BP-3 promoted the development of TH1-like TH17 cells under TH17-differentiating conditions, TH2-like T_reg cells occurred under T_reg polarization. Interestingly, differential effects could be observed in mixtures of the two tested compounds compared with the individual compounds. Notably, estrogen receptor β expression was decreased under TH17-differentiating conditions in the presence of BPA and BP-3 as mixture. In conclusion, our study provides solid evidence for both, the immune disruptive potential and the existence of cumulative effects of real nature EDC mixtures on T cell in vitro differentiation.

Activated partial thromboplastin time (aPTT) is a routine clotting assay that is widely used to globally screen for coagulation abnormalities, particularly in the pre-operative period. It is commonly admitted that a prolonged test result, may trigger the need for specific assays to be performed and particularly factor assay. However, the responsiveness of aPTT reagents to deficiencies of clotting factors varies because of differences in the type of activator and in the composition/concentration in phospholipids. To investigate the suitability of 5 commercially available aPTT reagents to detect mild/moderate deficiencies of clotting factors, we assessed their responsiveness at increasing concentrations of factors involved not only in the intrinsic pathway of the coagulation system, but also in the common pathway, as very few data were available. The sensitivity of 5 aPTT reagents i.e. HemosIL APTT SP (Instrumentation Laboratory, IL), HemosIL SynthASil (IL), STA-CK Prest (Stago), TriniCLOT aPTT HS (TCoag), and TriniCLOT Automated aPTT (TCoag) to clotting factor was assessed according to the recommendation of the CLSI H47-A2 guideline, by using factor-deficient plasmas spiked with a calibration plasma (all from IL) to produce individual FVIII:C, FIX, FXI, FXII, FV, FX, or FII activities ranging from <1% to ~100 %. Each of the spiked plasma samples was used for the determination of aPTT after being assayed to confirm the activity of the single factor. Tests were simultaneously performed in duplicate on the ACL TOP 700 analyzer (IL) and the values were averaged. Test results were expressed as the sample-to-control ratio, the latter was defined as the clotting time obtained in the calibration plasma containing ~100 % factor activity. The factor activity producing a prolongation of aPTT above 1.20 (patient-to-control ratio) was assigned as the factor responsiveness in % for that reagent. The level (in %) of a given clotting factor responsible for the prolongation of aPTT above 1.20 was highly variable from one aPTT reagent to another (Table). Moreover, for one given aPTT reagent, its sensitivity was very different depending on the specific factor. Actually, the aPTT responsiveness to FVIII:C ranged from 33 % with TriniCLOT aPTT HS to 46 % with HemosIL APTT SP. For FIX, the range was from 18 % with TriniCLOT Automated aPTT to 57 % with HemosIL SynthASil. The aPTT responsiveness to FXI ranged from 38 % with HemosIL SynthASil to 50 % with HemosIL APTT SP, and that to FXII was from 27 % with STA-CK Prest to 48 % with TriniCLOT aPTT HS. So, HemosIL APTT SP showed a good sensitivity (above 42%) to all 3 clotting factors which mild deficiencies are known to be associated with an hemorrhagic tendency (FVIII:C, FIX, and FXI). The same applied to HemosIL SynthASil, which was very sensitive to FIX deficiency (57%), and to STA-CK Prest and TriniCLOT aPTT HS with borderline sensitivity to FIX (29 % for both). The responsiveness to FIX of TriniCLOT Automated APTT was found to be very low (18%). Concerning coagulation factors involved in the common coagulation pathway, the sensitivity to FV was between 38 % with STA-CK Prest and 61 % with TriniCLOT Automated aPTT and that to FX was between 7.0 % with TriniCLOT aPTT HS and 49 % with HemosIL SynthASil. The sensitivity to FII was very low and quite similar for the 5 tested reagents in the range from 9.1 % to 10.7 %. The difference between reagents responsiveness to FIX was confirmed using plasma samples from patients with hemophilia B either treated with FIX concentrates or not. These results suggested that the sensitivity of the 5 tested aPTT reagents to single factor deficiency was highly variable. If the responsiveness to FVIII:C and FXI of the tested aPTT reagents was accurate, it was not the case for FIX with borderline sensitivity of STA-CK Prest and TriniCLOT aPTT HS (29 %), and more importantly a too low sensitivity of Triniclot Automated aPTT (18%). As this was confirm in clinical materials, this must be considered when analyzing clinical materials, particularly plasma from hemophilia B patients, as mild deficiency states might be undiagnosed with the less sensitive reagent.

Endocrine-disrupting chemicals such as benzophenone-3 (BP-3) can have severe consequences for human reproduction by affecting critical processes during pregnancy. To shed further light on potential harmful BP-3 actions, our current study addressed the impact of BP-3 on decidualization and trophoblast invasion. Decidualization was initiated in human endometrial stromal cells (THESC) upon treatment with a mixture of cAMP, progesterone, and estradiol. In parallel to hormonal treatment, the cells were exposed to different BP-3 concentrations ranging from 0.001 µM to 10 µM. The expression of decidualization and invasion markers was determined. Moreover, trophoblastic spheroids derived from JEG-3 cells were transferred to decidualized THESC after BP-3 exposure, and spheroid attachment and invasion were analyzed. Hormonal treatment successfully initiated decidualization in THESC, which was confirmed by increased prolactin levels and IGFBP1 and NCOA-3 mRNA expression. Notably, BP-3 exposure did not affect these markers. Furthermore, BP-3 changed neither THESC proliferation nor viability nor the frequency of cells expressing MMP2/9 or TIMP1/3. Trophoblastic spheroid attachment and outgrowth into THESC were not altered through any of the BP-3 concentrations applied. Our results do not provide evidence for an influence of BP-3 on the decidualization process and the capability of trophoblast cells to adhere and invade into endometrial stromal cells.

Zusammenfassung: Ob Bisphenole aus Plastikprodukten, per- und polyfluorierte Alkylsubstanzen (PFAS) aus Regenschutzkleidung oder Parabene aus Kosmetikprodukten – diese und weitere Chemikalien gelangen in jeden Haushalt und stellen eine potenzielle Gefahr für die Gesundheit dar, insbesondere während der sensiblen Phase der Schwangerschaft. Eine von uns durchgeführte Studie bestätigt den großen Bedarf an und den Wunsch von Frauen nach Aufklärung über Auswirkungen von Umweltchemikalien im Rahmen der Schwangerschaftsvorsorge.