Ikegami Akihiko, Chung Peter, Han Yiping W
Department of Periodontics, School of Dental Medicine, Case Western Reserve University, Cleveland, Ohio 44106-4905, USA.
Infect Immun. 2009 Jul;77(7):3075-9. doi: 10.1128/IAI.00209-09. Epub 2009 Apr 27.
Fusobacterium nucleatum is a gram-negative oral anaerobe implicated in periodontal disease and adverse pregnancy outcome. The organism colonizes the mouse placenta, causing localized infection and inflammation. The mechanism of placental colonization has not been elucidated. Previous studies identified a novel adhesin from F. nucleatum, FadA, as being involved in the attachment and invasion of host cells. The fadA deletion mutant F. nucleatum 12230 US1 was defective in host cell attachment and invasion in vitro, but it also exhibited pleiotropic effects with altered cell morphology and growth rate. In this study, a fadA-complementing clone, F. nucleatum 12230 USF81, was constructed. The expression of FadA on USF81 was confirmed by Western blotting and immunofluorescent labeling. USF81 restored host cell attachment and invasion activities. The ability of F. nucleatum 12230, US1, and USF81 to colonize the mouse placenta was examined. US1 was severely defective in placental colonization compared to the wild type and USF81. Thus, FadA plays an important role in F. nucleatum colonization in vivo. These results also represent the first complementation studies for F. nucleatum. FadA may be a therapeutic target for preventing F. nucleatum colonization of the host.
具核梭杆菌是一种革兰氏阴性口腔厌氧菌,与牙周疾病和不良妊娠结局有关。该菌可定殖于小鼠胎盘,引发局部感染和炎症。胎盘定殖的机制尚未阐明。先前的研究鉴定出一种来自具核梭杆菌的新型粘附素FadA,它参与宿主细胞的附着和侵袭。fadA缺失突变体具核梭杆菌12230 US1在体外宿主细胞附着和侵袭方面存在缺陷,但它也表现出多效性,细胞形态和生长速率发生了改变。在本研究中,构建了一个fadA互补克隆,具核梭杆菌12230 USF81。通过蛋白质免疫印迹和免疫荧光标记证实了USF81上FadA的表达。USF81恢复了宿主细胞的附着和侵袭活性。检测了具核梭杆菌12230、US1和USF81定殖于小鼠胎盘的能力。与野生型和USF81相比,US1在胎盘定殖方面存在严重缺陷。因此,FadA在具核梭杆菌体内定殖中起重要作用。这些结果也代表了对具核梭杆菌的首次互补研究。FadA可能是预防具核梭杆菌在宿主体内定殖的治疗靶点。