Sex Specific Expression of Interleukin 7, 8 and 15 in Placentas of Women with Gestational Diabetes. Keckstein Simon,Pritz Sophia,Amann Niklas,Meister Sarah,Beyer Susanne,Jegen Magdalena,Kuhn Christina,Hutter Stefan,Knabl Julia,Mahner Sven,Kolben Thomas,Jeschke Udo,Kolben Theresa M International journal of molecular sciences Gestational diabetes mellitus (GDM) is known to increase the risk for feto-maternal complications during pregnancy. A state of low-grade inflammation, with elevated levels of proinflammatory molecules, similar to patients with obesity or diabetes mellitus type 2 has also been partly described in GDM. The placenta, as unique interface between mother and fetus, is not only passively affected by changes in one of these organisms, but also acts as a modulator by expressing hormones and cytokines. This study aimed to investigate the expression of the proinflammatory cytokines Interleukin (IL) 7, 8 and 15 in GDM in placental tissue. A total number of 80 placentas were included (40 GDM/40 control group). The expression of IL-7, 8 and 15 was investigated in extravillous trophoblast (EVT) and syncytiotrophoblast (SCT) by immunohistochemistry and immunofluorescence double staining. The immunohistochemical staining was evaluated with the semiquanitfied immunoreactive score (IRS). While the expression IL-15 was significantly upregulated in EVTs of women with GDM. The expression of IL-8 was significantly decreased in EVT of the GDM group. Furthermore, significant fetal sex specific differences were detectable in all three cytokines. Our findings suggest an involvement of the investigated cytokines in the maintenance of a state of chronic low-grade inflammation on placental level in patients suffering from GDM. 10.3390/ijms21218026

Study of immune-tolerized cell lines and extracellular vesicles inductive environment promoting continuous expression and secretion of HLA-G from semiallograft immune tolerance during pregnancy. Journal of extracellular vesicles An immune reaction is a protector of our body but a target to be overcome for all non-self-derived medicine. Extracellular Vesicles (EVs), noted as a primary alternative to cell therapy products that exhibit immune rejection due to mismatching-major histocompatibility complex (MHC), were discovered to have excellent curative effects through the delivery of various biologically active substances. Although EVs are sure to incur immune reaction by immunogenicity due to alloantigens from their parental cells, their immune rejection is rarely known. Hence, to develop cell lines and EVs as medicines with no immune rejection, we noted the immune tolerance where the foetus, as semi-allograft, is perfectly protected from the maternal immune system. We designed the ex-vivo culture systems to simulate in-vivo environmental factors inducing extravillous trophoblast (EVT)-specific Human Leukocyte Antigen-G (HLA-G) expression and secretion of HLA-G-bearing EVs at the mother-foetus interface. Using them, we confirmed that immune-tolerized stem cells (itSCs) continuously expressing and secreting HLA-G like EVTs during pregnancy can be induced. Also, EVs secreted from itSCs are verified as immune-tolerized EVs (itSC-EVs) containing HLA-G and not causing immune rejection through various analytical methods. These findings can provide a new perspective on the local and extensive immune tolerance environment where HLA-G is expressed and secreted by pregnancy-related hormones and different biological conditions. Furthermore, they show the new way to develop itSCs-EVs-based therapeutics that are free from time, space, and donor limitation causing immune rejection. ABBREVIATIONS:CFSE: carboxyfluorescein succinimidyl ester; DC: dendritic cells; ELISA: enzyme-linked immunosorbent assay; EV: extracellular vesicles; EVT: extravillous trophoblast; FSH: follicle stimulating hormone; HA: hyaluronic acid; hCG: human chorionic gonadotropin; HLA-G: human leukocyte antigen G; iPSC: induced pluripotent stem cells; itSC-EVs: immune-tolerized extracellular vesicles from itSCs; itTBC-EVs: immune-tolerized extracellular vesicles from itTBCs; itSCs: immune tolerized stem cells; itTBCs: immune-tolerized trophoblast cells; LH: luteinizing hormone; MHC: major histocompatibility complex; MSC: mesenchymal stem cells; NK: natural killer cells; NTA: nanoparticle tracking analysis; PBMC: peripheral blood mononuclear cells; PHA: phytohemagglutinin; SP-IRIS: single particle interferometric reflectance imaging sensing; STB: syncytiotrophoblast. 10.1080/20013078.2020.1795364

IL-25 promotes trophoblast proliferation and invasion via binding with IL-17RB and associated with PE. Liu Siyu,Sun Yi,Tang Yao,Hu Rong,Zhou Qiongjie,Li Xiaotian Hypertension in pregnancy Interleukin-25 is a Th2 interleukin and has been shown to influence cell behavior. This study aims to illustrate its affection on extravillous trophoblasts function and association with PE. Methods qPCR and immunohistochemistry demonstrate IL-25 and IL-17RB expression. CCK-8 test and transwell were to access the behavior of HTR8 in the presence or absence of IL-25, IL-25 neutralization antibody. Results EVTs and HTR8 express IL-25 and IL-17RB. IL-25 promotes proliferation and invasion (p< 0.05),which was abolished in the presence of IL-25Ab (p< 0.05). Conclusion: IL-25 may contribute to promote trophoblast invasion and proliferation and abnormal decline of IL-25 might be associated with PE. qPCR and immunohistochemistry (IHC) were used to demonstrate IL-25 and its receptor IL-17RB expression in primary human trophoblasts of normal first- and third- trimester, as well as third-trimester of PE. CCK-8 test and transwell invasion system in vitro were applied separately to access the behavior of trophoblast cell line (HTR8) in the presence or absence of IL-25, IL-25 neutralization antibody (IL-25 Ab). EVTs and HTR8 express IL-25 and IL-17RB. The expressions increase in third-trimester during normal pregnancy. In PE, both of IL-25 and IL-17RB expressions decrease (< 0.05). IL-25 for 24 h promoted HTR8 proliferation and invasion (< 0.05). The effect was abolished in the presence of IL-25Ab (< 0.05). This study demonstrates that IL-25 may contribute to promote trophoblast invasion and proliferation and abnormal decline of IL-25 might be associated with PE. 10.1080/10641955.2021.1950177

Bisphenol S Impairs Invasion and Proliferation of Extravillous Trophoblasts Cells by Interfering with Epidermal Growth Factor Receptor Signaling. International journal of molecular sciences The placenta supports fetal growth and is vulnerable to exogenous chemical exposures. We have previously demonstrated that exposure to the emerging chemical bisphenol S (BPS) can alter placental endocrine function. Mechanistically, we have demonstrated that BPS interferes with epidermal growth factor receptor (EGFR) signaling, reducing placenta cell fusion. Extravillous trophoblasts (EVTs), a placenta cell type that aids with vascular remodeling, require EGF to invade into the maternal endometrium. We hypothesized that BPS would impair EGF-mediated invasion and proliferation in EVTs. Using human EVTs (HTR-8/SVneo cells), we tested whether BPS could inhibit the EGF response by blocking EGFR activation. We also evaluated functional endpoints of EGFR signaling, including EGF endocytosis, cell invasion and proliferation, and endovascular differentiation. We demonstrated that BPS blocked EGF-induced phosphorylation of EGFR by acting as a competitive antagonist to EGFR. Transwell assay and a three-dimensional microfluidic chip invasion assay revealed that BPS exposure can block EGF-mediated cell invasion. BPS also blocked EGF-mediated proliferation and endovascular differentiation. In conclusion, BPS can prevent EGF-mediated EVT proliferation and invasion through EGFR antagonism. Given the role of EGFR in trophoblast proliferation and differentiation during placental development, our findings suggest that maternal exposure to BPS may contribute to placental dysfunction via EGFR-mediated mechanisms. 10.3390/ijms23020671