Motedayyen Hossein, Fathi Farshid, Fasihi-Ramandi Mahdi, Sabzghabaee Ali Mohammad, Taheri Ramezan Ali
Nanobiotechnology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran.
Department of Immunology, Faculty of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran.
J Res Med Sci. 2019 Jan 31;24:1. doi: 10.4103/jrms.JRMS_463_18. eCollection 2019.
Maternal-fetal tolerance plays a fundamental role in the maintenance of pregnancy. However, this immunological tolerance can be influenced by intrauterine infections. Human amniotic epithelial cells (hAECs) have immunomodulatory effects and respond to invading pathogens through expressing various toll-like receptors (TLRs). We hypothesize that bacteria or bacterial products affect the immunosuppressive effects of hAECs through TLR stimulation. Here, we investigated how a successful pregnancy can be threatened by TLR4 activation on hAECs on lipopolysaccharide (LPS) engagement.
hAECs were isolated from the amniotic membrane received from six healthy pregnant women. The immunophenotyping of hAECs was studied by flow cytometry. The isolated hAECs (4 × 10 cells/ml) were cultured in 24-well plates in the presence or absence of LPS (5 μg/ml). After 24, 48, and 72 h of incubation, the culture supernatants of hAECs were collected, and the levels of interleukin-5 (IL-5), IL-6, IL-1β, tumor necrosis factor-alpha (TNF-α), transforming growth factor-beta 1 (TGF-β1), and prostaglandin E2 (PGE2) were measured by enzyme-linked immunosorbent assay.
TLR4 activation showed a stimulatory effect on TGF-β1 production of hAECs ( < 0.001-0.05). PGE2 production of LPS-stimulated hAECs was significantly increased ( < 0.01-0.05). Moreover, TLR4 could induce TNF-α and IL-1β production of hAECs ( < 0.0001-0.01), while this effect was not observed on IL-6 production of hAECs. The IL-5 was produced at a very low level in two culture supernatants of hAECs, in which its production was independent of LPS effect.
TLR4 activation by bacterial components on hAECs may be a potential risk factor for pregnancy complications.
母胎耐受在维持妊娠过程中起关键作用。然而，这种免疫耐受会受到宫内感染的影响。人羊膜上皮细胞（hAECs）具有免疫调节作用，并通过表达各种 Toll 样受体（TLRs）对入侵病原体作出反应。我们推测细菌或细菌产物通过 TLR 刺激影响 hAECs 的免疫抑制作用。在此，我们研究了脂多糖（LPS）作用下 TLR4 激活如何威胁 hAECs 上的成功妊娠。
从六名健康孕妇的羊膜中分离出 hAECs。通过流式细胞术研究 hAECs 的免疫表型。将分离的 hAECs（4×10 个细胞/毫升）在有或无 LPS（5 微克/毫升）的情况下培养于 24 孔板中。孵育 24、48 和 72 小时后，收集 hAECs 的培养上清液，并通过酶联免疫吸附测定法测量白细胞介素-5（IL-5）、IL-6、IL-1β、肿瘤坏死因子-α（TNF-α）、转化生长因子-β1（TGF-β1）和前列腺素 E2（PGE2）的水平。
TLR4 激活对 hAECs 的 TGF-β1 产生具有刺激作用（<0.001 - 0.05）。LPS 刺激的 hAECs 的 PGE2 产生显著增加（<0.01 - 0.05）。此外，TLR4 可诱导 hAECs 的 TNF-α 和 IL-1β 产生（<0.0001 - 0.01），而在 hAECs 的 IL-6 产生上未观察到这种作用。hAECs 的两种培养上清液中 IL-5 的产生水平非常低，其产生与 LPS 效应无关。
细菌成分对 hAECs 上 TLR4 的激活可能是妊娠并发症的一个潜在危险因素。
