Pathogenic Role of Associated Adherent-Invasive Escherichia coli in Crohn's Disease
Pathogenic Role of Associated Adherent-Invasive Escherichia coli in Crohn's Disease
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DOI：
10.1002/jcp.25717
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发表时间：
2017-10-01
影响因子：
5.6
通讯作者：
De Luca, Antonio
中科院分区：
文献类型：
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作者：
Mazzarella, Giuseppe;Perna, Angelica;De Luca, Antonio
一些证据表明，粘附-侵袭性大肠杆菌（AIEC）菌株在克罗恩病（CD）中起重要作用。本研究的目的是探讨两种AIEC菌株LF82和O83:H1在CD患者中的致病作用。建立患者结肠活检组织的器官培养，通过免疫组织化学方法评估LF82和O83: H1对CEACAM6、LAMP1、HLA-DR、ICAM1基因表达的影响，通过RT-PCR方法评估IL-8、IFN γ和tnf - α基因表达的影响。此外，在Caco2细胞上，我们分别通过FACS、RT-PCR和DAPI染色分析了细胞周期、MGMT和DNMT1基因的表达以及LF82和O83: H1诱导的DNA损伤。上皮和固有层单核细胞（LPMNC） CEACAM6和LAMP1的表达在O83: H1和LF82培养的活检组织中高于非致病性大肠杆菌培养的活检组织。两种AIEC菌株均诱导血管上ICAM-1和LPMNC上HLA-DR的表达增加。我们观察到，两种AIEC菌株培养的活检组织中tnf - α、ifn - γ和IL-8转录物的水平高于NP培养的活检组织。LF82和O83: H1阻断细胞周期进入S期，诱导DNA损伤，调节DNMT1和MGMT基因的表达。我们的数据表明，大肠杆菌LF82和083:H1菌株能够增加CD结肠活检中所有促炎细胞因子和所有粘膜免疫标记物的表达。
Several lines of evidence suggest that adherent-invasive Escherichia coli (AIEC) strains play an important role in Crohn's disease (CD). The objective of this study was to investigate the pathogenic role of two AIEC strains, LF82 and O83:H1, in CD patients. Organ cultures of colonic biopsies from patients were set up to assess the effects of LF82 and O83: H1 on the expression of CEACAM6, LAMP1, HLA-DR, ICAM1 by immunohistochemistry and of IL-8, IFN gamma , and TNF-alpha genes by RT-PCR. Moreover, on Caco2 cells, we analyzed the cell cycle, the expression of MGMT and DNMT1 genes, and DNA damage induced by LF82 and O83: H1, by FACS, RT-PCR, and DAPI staining, respectively. Epithelial and lamina propria mononuclear cells (LPMNC) expression of CEACAM6 and LAMP1 were higher in biopsies cultured in the presence of both O83: H1 and LF82 than in biopsies cultured with non-pathogenic E. coli. Both AIEC strains induced increased expression of ICAM-1 on blood vessels and HLA-DR on LPMNC. We observed higher levels of TNF-alpha, IFN-gamma, and IL-8 transcripts in biopsies cultured with both AIEC strains than in those cultured with NP. Both LF82 and O83: H1, block the cell cycle into S phase, inducing DNA damage, and modulate the expression of DNMT1 and MGMT genes. Our data suggest that LF82 and 083: H1 strains of E. coli are able to increase in CD colonic biopsies the expression of all the pro-inflammatory cytokines and all the mucosal immune markers investigated.