Ohkusa Toshifumi, Yoshida Tsutomu, Sato Nobuhiro, Watanabe Sumio, Tajiri Hisao, Okayasu Isao
Department of Gastroenterology and Hepatology, The Jikei University School of Medicine, Kashiwa, Chiba 277-8567, Japan.
Department of Pathology, Kitasato University School of Medicine, Sagamihara, Kanagawa 228-8555, Japan.
J Med Microbiol. 2009 May;58(Pt 5):535-545. doi: 10.1099/jmm.0.005801-0.
Interleukin 2 (IL-2)- and IL-10-knockout mice develop spontaneous colitis under conventional but not germ-free conditions, suggesting that commensal bacteria play an important role in the pathogenesis of colitis. However, interactions between commensal bacteria and colonic epithelial cells have not been fully investigated. We therefore assessed the ability of various commensal bacteria and probiotics to adhere to and invade colonic epithelial cells. Effects of the bacteria on production of proinflammatory cytokines were also measured. Commensal bacteria, including mucosal organisms isolated from ulcerative colitis (UC) patients, such as Fusobacterium varium, reported as a possible pathogen in UC, Bacteroides vulgatus, Escherichia coli and Clostridium clostridioforme, as well as their type strains and probiotics, were assessed for their ability to adhere to and invade colonic epithelial cells using two cell lines, SW-480 and HT-29. Our experiments employed co-incubation, a combination of scanning and transmission electron microscopy and recovery of bacteria from infected-cell lysates. F. varium and several other commensal bacteria, but not probiotics, adhered to colonic epithelial cells and invaded their cytoplasm. ELISA and real-time PCR revealed that the host cells, particularly those invaded by F. varium, showed significant increases in IL-8 and TNF-alpha concentrations in supernatants, with elevation of IL-8, TNF-alpha, MCP-1 and IL-6 mRNAs. Furthermore, IL-8 and TNF-alpha expression and nuclear phosphorylated NF-kappaB p65 expression could be immunohistochemically confirmed in inflamed epithelium with cryptitis or crypt abscess in UC patients. Certain commensal bacteria can invade colonic epithelial cells, activating early intracellular signalling systems to trigger host inflammatory reactions.
白细胞介素2(IL-2)基因敲除小鼠和白细胞介素10基因敲除小鼠在常规条件下而非无菌条件下会发生自发性结肠炎,这表明共生菌在结肠炎的发病机制中起重要作用。然而,共生菌与结肠上皮细胞之间的相互作用尚未得到充分研究。因此,我们评估了各种共生菌和益生菌黏附并侵入结肠上皮细胞的能力。还检测了这些细菌对促炎细胞因子产生的影响。使用SW-480和HT-29两种细胞系,评估了共生菌,包括从溃疡性结肠炎(UC)患者分离出的黏膜微生物,如据报道可能是UC病原体的多变梭杆菌、普通拟杆菌、大肠杆菌和梭状芽孢杆菌样梭菌,以及它们的模式菌株和益生菌黏附并侵入结肠上皮细胞的能力。我们的实验采用了共孵育、扫描和透射电子显微镜相结合以及从感染细胞裂解物中回收细菌的方法。多变梭杆菌和其他几种共生菌而非益生菌黏附于结肠上皮细胞并侵入其细胞质。酶联免疫吸附测定(ELISA)和实时聚合酶链反应(PCR)显示,宿主细胞,尤其是被多变梭杆菌侵入的细胞,其培养上清液中白细胞介素8(IL-8)和肿瘤坏死因子α(TNF-α)浓度显著升高,同时IL-8、TNF-α、单核细胞趋化蛋白-1(MCP-1)和白细胞介素6(IL-6)的信使核糖核酸(mRNA)水平升高。此外,在UC患者有隐窝炎或隐窝脓肿的炎症上皮中,可通过免疫组织化学证实IL-8和TNF-α的表达以及核磷酸化核因子κB p65(NF-κB p65)的表达。某些共生菌可侵入结肠上皮细胞,激活早期细胞内信号系统以引发宿主炎症反应。