Conte Maria P, Aleandri Marta, Marazzato Massimiliano, Conte Antonietta L, Ambrosi Cecilia, Nicoletti Mauro, Zagaglia Carlo, Gambara Guido, Palombi Fioretta, De Cesaris Paola, Ziparo Elio, Palamara Anna T, Riccioli Anna, Longhi Catia
Department of Public Health and Infectious Diseases, Sapienza University, Rome, Italy
Department of Public Health and Infectious Diseases, Sapienza University, Rome, Italy.
Infect Immun. 2016 Oct 17;84(11):3105-3113. doi: 10.1128/IAI.00438-16. Print 2016 Nov.
Adherent/invasive Escherichia coli (AIEC) strains have recently been receiving increased attention because they are more prevalent and persistent in the intestine of Crohn's disease (CD) patients than in healthy subjects. Since AIEC strains show a high percentage of similarity to extraintestinal pathogenic E. coli (ExPEC), neonatal meningitis-associated E. coli (NMEC), and uropathogenic E. coli (UPEC) strains, here we compared AIEC strain LF82 with a UPEC isolate (strain EC73) to assess whether LF82 would be able to infect prostate cells as an extraintestinal target. The virulence phenotypes of both strains were determined by using the RWPE-1 prostate cell line. The results obtained indicated that LF82 and EC73 are able to adhere to, invade, and survive within prostate epithelial cells. Invasion was confirmed by immunofluorescence and electron microscopy. Moreover, cytochalasin D and colchicine strongly inhibited bacterial uptake of both strains, indicating the involvement of actin microfilaments and microtubules in host cell invasion. Moreover, both strains belong to phylogenetic group B2 and are strong biofilm producers. In silico analysis reveals that LF82 shares with UPEC strains several virulence factors: namely, type 1 pili, the group II capsule, the vacuolating autotransporter toxin, four iron uptake systems, and the pathogenic island (PAI). Furthermore, compared to EC73, LF82 induces in RWPE-1 cells a marked increase of phosphorylation of mitogen-activated protein kinases (MAPKs) and of NF-κB already by 5 min postinfection, thus inducing a strong inflammatory response. Our in vitro data support the hypothesis that AIEC strains might play a role in prostatitis, and, by exploiting host-cell signaling pathways controlling the innate immune response, likely facilitate bacterial multiplication and dissemination within the male genitourinary tract.
黏附/侵袭性大肠杆菌(AIEC)菌株最近受到越来越多的关注,因为它们在克罗恩病(CD)患者肠道中比在健康受试者中更普遍且更持久。由于AIEC菌株与肠外致病性大肠杆菌(ExPEC)、新生儿脑膜炎相关大肠杆菌(NMEC)及尿路致病性大肠杆菌(UPEC)菌株有很高的相似性,我们在此将AIEC菌株LF82与一株UPEC分离株(菌株EC73)进行比较,以评估LF82是否能够作为肠外靶标感染前列腺细胞。通过使用RWPE - 1前列腺细胞系来确定两种菌株的毒力表型。所得结果表明,LF82和EC73能够黏附、侵袭前列腺上皮细胞并在其中存活。通过免疫荧光和电子显微镜证实了侵袭。此外,细胞松弛素D和秋水仙碱强烈抑制两种菌株的细菌摄取,表明肌动蛋白微丝和微管参与宿主细胞侵袭。此外,两种菌株都属于系统发育B2组,并且是强生物膜生产者。计算机分析表明,LF82与UPEC菌株共有几种毒力因子:即1型菌毛、II组荚膜、空泡自转运毒素、四种铁摄取系统和致病岛(PAI)。此外,与EC73相比,LF82在感染后5分钟即可诱导RWPE - 1细胞中丝裂原活化蛋白激酶(MAPK)和NF-κB的磷酸化显著增加,从而引发强烈的炎症反应。我们的体外数据支持这样的假设,即AIEC菌株可能在前列腺炎中起作用,并且通过利用控制先天免疫反应的宿主细胞信号通路,可能促进细菌在男性泌尿生殖道内的繁殖和传播。