Amabebe Emmanuel, Richardson Lauren S, Kumar Awanit, Menon Ramkumar, Taylor Brandie D
Department of Obstetrics and Gynecology, University of Texas Medical Branch at Galveston, Galveston, Texas, USA.
Advocate Aurora Research Institute, Milwaukee, Wisconsin, USA.
Am J Reprod Immunol. 2025 Sep;94(3):e70129. doi: 10.1111/aji.70129.
We tested the hypothesis that oxidative stress (OS)-induced inflammatory response in decidual cells (DECs) may transfer and/or trigger the release of interferon epsilon (IFNε)-positive extracellular vesicles (EVs) from vaginal epithelial cells (VECs) to minimize vaginal disturbances.
VECs were treated for 48 h under the following conditions: (1) standard VEC media, (2) OS-inducing cigarette smoke extract (CSE); and supernatant from (3) normal/untreated DECs, and (4) CSE-treated DECs. The concentration of cytoplasmic, secreted, and VEC-derived EV bound IFNε (n = 3 each) was measured by enzyme-linked immunosorbent assay (ELISA). EVs were isolated from culture media by cushioned-density gradient ultracentrifugation and characterized by immunoblotting and nanoparticle tracking analysis.
Induction of OS in VECs with CSE increased intracellular IFNε in VECs (p = 0.0007) but not free or EV-bound IFNε compared to control VECs. Exposure to conditioned media from untreated and CSE-treated DECs induced increased intracellular (p = 0.004, p = 0.049) and free IFNε (p = 0.04, p = 0.03) from VECs. VEC-derived EVs (126 ± 11.8 nm) expressed exosome markers, and did not change in size regardless of the treatment but decreased in number due to exposure to untreated (p = 0.004) and CSE-treated DECs (p = 0.025) conditioned media. Furthermore, VEC exosomal IFNε increased by 2.6-fold (p = 0.0001, untreated DECs) and ∼4-fold (p = 0.041, CSE-treated DECs) compared to controls.
Mucosal immune defense mediated by IFNε may be an innate response by VECs under OS. This was further evidenced by an overall increase in IFNε due to both physiologic and pathologic impact of decidua on VECs. IFNε may indicate a stress response by VECs or paracrine crosstalk between gestational tissues.
我们检验了以下假设：蜕膜细胞（DEC）中氧化应激（OS）诱导的炎症反应可能会转移和/或触发阴道上皮细胞（VEC）释放干扰素ε（IFNε）阳性细胞外囊泡（EV），以尽量减少阴道紊乱。
VEC在以下条件下处理48小时：（1）标准VEC培养基，（2）诱导OS的香烟烟雾提取物（CSE）；以及来自（3）正常/未处理的DEC和（4）经CSE处理的DEC的上清液。通过酶联免疫吸附测定（ELISA）测量细胞质、分泌的和VEC衍生的EV结合的IFNε的浓度（每组n = 3）。通过缓冲密度梯度超速离心从培养基中分离EV，并通过免疫印迹和纳米颗粒跟踪分析进行表征。
与对照VEC相比，用CSE诱导VEC中的OS会增加VEC中的细胞内IFNε（p = 0.0007），但不会增加游离或EV结合的IFNε。暴露于未处理和经CSE处理的DEC的条件培养基会诱导VEC中细胞内（p = 0.004，p = 0.049）和游离IFNε（p = 0.04，p = 0.03）增加。VEC衍生的EV（126±11.8nm）表达外泌体标志物，无论处理如何，其大小均无变化，但由于暴露于未处理（p = 0.004）和经CSE处理的DEC（p = 0.025）的条件培养基中，其数量减少。此外，与对照相比，VEC外泌体IFNε增加了2.6倍（p = 0.0001，未处理的DEC）和约4倍（p = 0.041，经CSE处理的DEC）。
IFNε介导的黏膜免疫防御可能是VEC在OS下的一种固有反应。蜕膜对VECs的生理和病理影响导致IFNε总体增加，进一步证明了这一点。IFNε可能表明VEC的应激反应或妊娠组织之间的旁分泌串扰。
