Probiotic Lactobacilli and Vaginal Epithelium

描述（由申请人提供）：尿路感染（UTI）最常见由大肠埃希菌引起，在所有年龄段的女性中很常见。最近在尿路病原体中抗菌素耐药性的增加刺激了对替代的、非抗菌素的方法代替抗生素来预防和治疗UTI的兴趣。UTI发生之前，阴道植物群会发生特异性改变。通常由乳杆菌属物种占主导地位，阴道微粒体被E.在UTI发展之前，阴道乳酸杆菌的消耗与E.大肠杆菌和其他病原体，增加了UTI和某些性传播疾病的风险。一项正在进行的临床研究正在调查阴道乳杆菌栓剂，卷曲乳杆菌CTV-05，在预防复发性UTI（DK 53369）的疗效。然而，尚未系统地研究阴道中乳酸杆菌益生菌株介导保护的机制。该建议的中心假设是阴道乳杆菌如该益生菌分离物卷曲乳杆菌CTV-05利用特定机制保护阴道上皮免受病原体如尿路致病性大肠杆菌的侵害。杆菌我们将使用L.卷曲肠杆菌CTV-05、其他分离株和原代培养的阴道上皮细胞：（1）乳酸杆菌利用特异性粘附分子粘附于阴道上皮，并保护免受尿路致病性大肠杆菌的粘附。大肠杆菌通过竞争性抑制，包括通过结合阴道上皮细胞（VEC）上的常见球状系列GSL受体。粘附也可能通过S层介导;（2）乳酸杆菌杀死尿路致病性E。通过产生HZO_2和类细菌素抑制尿路致病性大肠杆菌的生长。coli共聚集;与大肠杆菌等病原体相比，乳酸杆菌在粘附于阴道上皮时诱导修复和屏障功能基因。大肠杆菌，诱导促炎基因。这些研究将对理解阴道微环境和其他上皮表面（如胃肠道和上呼吸道）中正常植物群和益生菌的保护机制具有重要意义，并将建议合理设计和/或增强益生菌。

DESCRIPTION (provided by applicant): Urinary tract infections (UTIs) are most frequently caused by Escherichia coli and are common among women of all ages. Recent increases in antimicrobial resistance among uropathogens have stimulated interest in alternative, non-antimicrobial methods of preventing and treating UTI in lieu of antibiotics. UTI is preceded by specific alterations in the vaginal flora. Normally dominated by Lactobacillus species, the vaginal microniche becomes colonized with E. coli prior to the development of UTI. Depletion of vaginal lactobacilli is associated with vaginal colonization with E. coli and other pathogens, increasing the risk of UTI and certain sexually transmitted diseases. An ongoing clinical study is investigating the efficacy of a vaginal Lactobacillus suppository, L crispatus CTV-05, in preventing recurrent UTI (DK53369). However, the mechanisms that mediate protection by probiotic strains of Lactobacillus in the vagina have not been systematically investigated. The central hypothesis of this proposal is that vaginal lactobacilli such as this probiotic isolate L crispatus CTV-05 utilize specific mechanisms in protecting the vaginal epithelium against pathogens such as uropathogenic E. coli. We will investigate the following specific hypotheses, using L. crispatus CTV-05, additional isolates from our collections, and primary cultured vaginal epithelial cells: (1) Lactobacilli adhere to the vaginal epithelium using specific adherence molecules and protect against adherence of uropathogenic E. coli via competitive inhibition, including by binding to common globoseries GSL receptors on vaginal epithelial cells (VECs). Adherence may also be mediated via S-layers; (2) Lactobacilli kill uropathogenic E. coli in vitro via production of HZO2 and bacteriocin-like substances and inhibits growth of uropathogenic E. coli via co-aggregation; and (3) Lactobacilli induce repair and barrier function genes upon adherence to vaginal epithelium, in contrast with pathogens such as E. coli, which induce proinflammatory genes. These studies will have important implications for understanding mechanisms of protection by normal flora and probiotics in the vaginal microniche and at other epithelial surfaces such as the gastrointestinal and upper respiratory tracts and will suggest rational design and/or enhancement of probiotics.