Center for Research on Reproduction and Women's Health, Department of Obstetrics and Gynecology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA; Department of Microbiology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
Center for Research on Reproduction and Women's Health, Department of Obstetrics and Gynecology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
J Reprod Immunol. 2022 Aug;152:103648. doi: 10.1016/j.jri.2022.103648. Epub 2022 May 23.
Lactobacillus-deficient cervicovaginal microbiota, including Gardnerella vaginalis, are implicated in cervical remodeling and preterm birth. Mechanisms by which microbes drives outcomes are not fully elucidated. We hypothesize that Gardnerella vaginalis induces matrix metalloproteinases through TLR-2, leading to epithelial barrier dysfunction and premature cervical remodeling. Cervicovaginal cells were treated with live Gardnerella vaginalis or Lactobacillus crispatus or their bacteria-free supernatants for 24 h. For TLR-2 experiments, cells were pretreated with TLR-2 blocking antibody. A Luminex panel was run on cell media. For human data, we conducted a case-control study from a prospective pregnancy cohort of Black individuals with spontaneous preterm (sPTB) (n = 40) or term (n = 40) births whose vaginal microbiota had already been characterized. Cervicovaginal fluid was obtained between 20 and 24 weeks' gestation. Short cervix was defined as < 25 mm by second trimester transvaginal ultrasound. MMP-9 was quantified by ELISA. Standard analytical approaches were used to determine differences across in vitro conditions, as well as MMP-9 and associations with clinical outcomes. Gardnerella vaginalis induced MMP-1 in cervical cells (p = 0.01) and MMP-9 in cervical and vaginal (VK2) cells (p ≤ 0.001 for all). TLR-2 blockade mitigated MMP-9 induction by Gardnerella vaginalis. MMP-9 in cervicovaginal fluid is higher among pregnant individuals with preterm birth, short cervix, and Lactobacillus-deficient microbiota (p < 0.05 for all). MMP-9 is increased in the cervicovaginal fluid of pregnant individuals with subsequent sPTB. Our in vitro work ascribes a potential mechanism by which a cervicovaginal microbe, commonly associated with adverse pregnancy outcomes, may disrupt the cervicovaginal epithelial barrier and promote premature cervical remodeling in spontaneous preterm birth.
阴道缺乏乳杆菌的阴道微生物群,包括阴道加德纳菌,与宫颈重塑和早产有关。微生物驱动结局的机制尚未完全阐明。我们假设阴道加德纳菌通过 TLR-2 诱导基质金属蛋白酶,导致上皮屏障功能障碍和早产宫颈重塑。将宫颈阴道细胞用活的阴道加德纳菌或卷曲乳杆菌或其无细菌上清液处理 24 小时。对于 TLR-2 实验,细胞用 TLR-2 阻断抗体预处理。细胞培养基上运行 Luminex 面板。对于人类数据,我们对来自黑种人自发性早产(sPTB)(n=40)或足月(n=40)分娩的前瞻性妊娠队列进行了病例对照研究,这些人的阴道微生物群已经得到了描述。在妊娠 20-24 周时获得宫颈阴道液。通过经阴道超声在妊娠中期将短颈定义为<25mm。通过 ELISA 定量 MMP-9。采用标准分析方法来确定体外条件下的差异,以及 MMP-9 与临床结局的关系。阴道加德纳菌诱导宫颈细胞中的 MMP-1(p=0.01)和宫颈和阴道(VK2)细胞中的 MMP-9(p≤0.001 所有)。TLR-2 阻断减轻了阴道加德纳菌诱导的 MMP-9 诱导。早产、短颈和乳杆菌缺乏的孕妇阴道液中的 MMP-9 较高(所有 p<0.05)。随后发生 sPTB 的孕妇阴道液中的 MMP-9 增加。我们的体外研究归因于一种潜在的机制,即与不良妊娠结局相关的阴道微生物可能破坏宫颈阴道上皮屏障并促进自发性早产的早产宫颈重塑。