Strömbeck Louise, Sandros Jens, Holst Elisabeth, Madianos Phoebus, Nannmark Ulf, Papapanou Panos, Mattsby-Baltzer Inger
Department of Clinical Bacteriology, Göteborg University, Göteborg, Sweden.
APMIS. 2007 Mar;115(3):241-51. doi: 10.1111/j.1600-0463.2007.apm_512.x.
Prevotella bivia has been associated with female upper genital tract infections and an increased risk of preterm delivery. In this study, the adherence and invasion capacity of P. bivia was investigated using a cervix epithelial cell line. P. bivia was furthermore analysed for its ability to evoke a proinflammatory cytokine response in epithelial cells. The invasion capacity, defined as the number of bacteria recovered from lysed HeLa cells infected with P. bivia, varied considerably among five strains, all of which were isolates from women with bacterial vaginosis. One P. bivia strain (P47) gave rise to an approximately 120-fold higher number of intracellular bacteria (7 x 10(3) bacteria per 1 x 10(5) cells) compared with the least invasive strain. Three strains expressed an intermediate or low invasiveness, showing an approximately 3- to 40-fold higher number of intracellular bacteria per 1 x 10(5) cells compared with the least invasive strain. The intracellular localization of P47 in phagosome-like vesicles was confirmed by transmission electron microscopy. All P. bivia strains adhered to HeLa cells to the same extent (range 14-22 bacteria per cell) as analysed by interference microscopy. No correlation was found between adhesion and invasion capacity of the strains. Furthermore, no fimbriae-like structures were observed on P47 detected by scanning electron microscopy or negative staining. Analysis of TNF-alpha, IL-1alpha, IL-6, IL-8, and IL-18 in P. bivia-stimulated HeLa cells showed low levels of only IL-6 and IL-8 for the most invasive P. bivia strain P47. Thus, the induction of IL-6 or IL-8 secretion appeared to be associated with invasion capacity. This work provides evidence that some P. bivia isolates can invade human cervix epithelial. Thus, a strong capacity for invasion and a weak proinflammatory cytokine-inducing capacity in P. bivia are suggested to be virulence factors in establishing a low-grade upper genital tract infection.
二路普雷沃菌与女性上生殖道感染及早产风险增加有关。在本研究中,使用宫颈上皮细胞系研究了二路普雷沃菌的黏附与侵袭能力。此外,还分析了二路普雷沃菌在上皮细胞中引发促炎细胞因子反应的能力。侵袭能力定义为从感染二路普雷沃菌的裂解海拉细胞中回收的细菌数量,在五株菌株中差异很大,所有菌株均分离自细菌性阴道病女性。与侵袭性最低的菌株相比,一株二路普雷沃菌菌株(P47)产生的细胞内细菌数量高出约120倍(每1×10⁵个细胞中有7×10³个细菌)。三株菌株表现出中等或低侵袭性,与侵袭性最低的菌株相比,每1×10⁵个细胞中的细胞内细菌数量高出约3至40倍。通过透射电子显微镜证实了P47在吞噬体样小泡中的细胞内定位。通过干涉显微镜分析,所有二路普雷沃菌菌株对海拉细胞的黏附程度相同(每细胞14 - 22个细菌)。未发现菌株的黏附与侵袭能力之间存在相关性。此外,通过扫描电子显微镜或负染色在P47上未观察到类似菌毛的结构。对二路普雷沃菌刺激的海拉细胞中肿瘤坏死因子-α、白细胞介素-1α、白细胞介素-6、白细胞介素-8和白细胞介素-18的分析表明,侵袭性最强的二路普雷沃菌菌株P47仅产生低水平的白细胞介素-6和白细胞介素-8。因此,白细胞介素-6或白细胞介素-8分泌的诱导似乎与侵袭能力有关。这项工作提供了证据表明一些二路普雷沃菌分离株可以侵入人宫颈上皮。因此,二路普雷沃菌强大的侵袭能力和较弱的促炎细胞因子诱导能力被认为是导致低度上生殖道感染的毒力因子。