Amabebe Emmanuel, Richardson Lauren S, Bento Giovana Fernanda Cosi, Radnaa Enkhtuya, Kechichian Talar, Menon Ramkumar, Anumba Dilly O C
Department of Oncology and Metabolism, University of Sheffield, Sheffield, UK.
Department of Obstetrics and Gynecology, The University of Texas Medical Branch at Galveston, Galveston, TX, USA.
Mol Biol Rep. 2023 Apr;50(4):3035-3043. doi: 10.1007/s11033-022-08183-6. Epub 2023 Jan 20.
Ureaplasma, a genus of the order Mycoplasmatales and commonly grouped with Mycoplasma as genital mycoplasma is one of the most common microbes isolated from women with infection/inflammation-associated preterm labor (PTL). Mycoplasma spp. produce sialidase that cleaves sialic acid from glycans of vaginal mucous membranes and facilitates adherence and invasion of the epithelium by pathobionts, and dysregulated immune response. However, whether Ureaplasma species can induce the production of sialidase is yet to be demonstrated. We examined U. parvum-infected vaginal epithelial cells (VECs) for the production of sialidase and pro-inflammatory cytokines.
Immortalized VECs were cultured in appropriate media and treated with U. parvum in a concentration of 1 × 10 DNA copies/ml. After 24 h of treatment, cells and media were harvested. To confirm infection and cell uptake, immunocytochemistry for multi-banded antigen (MBA) was performed. Pro-inflammatory cytokine production and protein analysis for sialidase confirmed pro-labor pathways.
Infection of VECs was confirmed by the presence of intracellular MBA. Western blot analysis showed no significant increase in sialidase expression from U. parvum-treated VECs compared to uninfected cells. However, U. parvum infection induced 2-3-fold increased production of GM-CSF (p = 0.03), IL-6 (p = 0.01), and IL-8 (p = 0.01) in VECs compared to controls.
U. parvum infection of VECs induced inflammatory imbalance associated with vaginal dysbiosis but did not alter sialidase expression at the cellular level. These data suggest that U. parvum's pathogenic effect could be propagated by locally produced pro-inflammatory cytokines and, unlike other genital mycoplasmas, may be independent of sialidase.
脲原体是支原体目下的一个属，通常与支原体一起被归类为生殖支原体，是从感染/炎症相关的早产（PTL）女性中分离出的最常见微生物之一。支原体属产生唾液酸酶，该酶可从阴道黏膜聚糖中裂解唾液酸，并促进致病共生菌对上皮的黏附和侵袭，以及免疫反应失调。然而，脲原体是否能诱导唾液酸酶的产生尚未得到证实。我们检测了微小脲原体感染的阴道上皮细胞（VECs）中唾液酸酶和促炎细胞因子的产生情况。
将永生化的VECs培养在合适的培养基中，并用浓度为1×10 DNA拷贝/ml的微小脲原体处理。处理24小时后，收集细胞和培养基。为了确认感染和细胞摄取情况，进行了多带抗原（MBA）的免疫细胞化学检测。促炎细胞因子的产生和唾液酸酶的蛋白质分析证实了促分娩途径。
通过细胞内MBA的存在证实了VECs的感染。蛋白质印迹分析显示，与未感染的细胞相比，微小脲原体处理的VECs中唾液酸酶表达没有显著增加。然而，与对照组相比，微小脲原体感染诱导VECs中GM-CSF（p = 0.03）、IL-6（p = 0.01）和IL-8（p = 0.01）的产生增加了2 - 3倍。
VECs的微小脲原体感染诱导了与阴道生态失调相关的炎症失衡，但在细胞水平上没有改变唾液酸酶的表达。这些数据表明，微小脲原体的致病作用可能由局部产生的促炎细胞因子介导，并且与其他生殖支原体不同，可能独立于唾液酸酶。
