Kim Sandra C, Tonkonogy Susan L, Karrasch Thomas, Jobin Christian, Sartor R Balfour
Center for Gastrointestinal Biology and Disease, University of North Carolina, Chapel Hill, NC, USA.
Inflamm Bowel Dis. 2007 Dec;13(12):1457-66. doi: 10.1002/ibd.20246.
Monoassociating gnotobiotic IL-10-deficient (-/-) mice with either nonpathogenic Enterococcus faecalis or a nonpathogenic Escherichia coli strain induces T-cell-mediated colitis with different kinetics and anatomical location (E. faecalis: late onset, distal colonic; E. coli: early onset, cecal).
E. faecalis and E. coli act in an additive manner to induce more aggressive colitis than disease induced by each bacterial species independently.
Germ-free (GF) inbred 129S6/SvEv IL-10-/- and wildtype (WT) mice inoculated with nonpathogenic E. faecalis and/or E. coli were killed 3-7 weeks later. Colonic segments were scored histologically for inflammation (0 to 4) or incubated in media overnight to measure spontaneous IL-12/IL-23p40 secretion. Bacterial species were quantified by serial dilution and plated on culture media. Mesenteric lymph node (MLN) CD4(+) cells were stimulated with antigen-presenting cells pulsed with bacterial lysate (E. faecalis, E. coli, Bacteroides vulgatus) or KLH (unrelated antigen control). IFN-gamma and IL-17 levels were measured in the supernatants.
Dual-associated IL-10-/- (but not WT) mice developed mild-to-moderate pancolitis by 3 weeks that progressed to severe distal colonic-predominant pancolitis with reactive atypia and duodenal inflammation by 7 weeks. NF-kappaB was activated in the duodenum and colon in dual-associated IL-10-/- x NF-kappaB(EGFP) mice. The aggressiveness of intestinal inflammation and the degree of antigen-specific CD4(+) cell activation were greater in dual- versus monoassociated IL-10-/- mice.
Two commensal bacteria that individually induce phenotypically distinct colitis in gnotobiotic IL-10-/- mice act additively to induce aggressive pancolitis and duodenal inflammation.
将无菌的白细胞介素-10缺陷（-/-）悉生小鼠单独与非致病性粪肠球菌或非致病性大肠杆菌菌株定殖，会诱导出具有不同动力学和解剖位置的T细胞介导的结肠炎（粪肠球菌：发病晚，结肠远端；大肠杆菌：发病早，盲肠）。
粪肠球菌和大肠杆菌以累加方式起作用，诱导出比每种细菌单独诱导的疾病更具侵袭性的结肠炎。
3至7周后处死接种了非致病性粪肠球菌和/或大肠杆菌的无菌（GF）近交129S6/SvEv白细胞介素-10-/-和野生型（WT）小鼠。对结肠段进行组织学炎症评分（0至4），或在培养基中孵育过夜以测量自发白细胞介素-12/白细胞介素-23p40分泌。通过系列稀释对细菌种类进行定量，并接种于培养基上。用脉冲了细菌裂解物（粪肠球菌、大肠杆菌、脆弱拟杆菌）或钥孔戚血蓝蛋白（无关抗原对照）的抗原呈递细胞刺激肠系膜淋巴结（MLN）CD4(+)细胞。测量上清液中的干扰素-γ和白细胞介素-17水平。
到3周时，双重定殖的白细胞介素-10-/-（而非WT）小鼠出现轻度至中度全结肠炎，到7周时进展为严重的以远端结肠为主的全结肠炎，并伴有反应性异型增生和十二指肠炎症。在双重定殖的白细胞介素-10-/-×核因子-κB（增强绿色荧光蛋白）小鼠的十二指肠和结肠中，核因子-κB被激活。与单一细菌定殖的白细胞介素-10-/-小鼠相比，双重细菌定殖的白细胞介素-10-/-小鼠肠道炎症的侵袭性和抗原特异性CD4(+)细胞激活程度更高。
在无菌白细胞介素-10-/-小鼠中，两种分别诱导表型不同结肠炎的共生细菌以累加方式起作用，诱导出侵袭性全结肠炎和十二指肠炎症。
