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首页> 外文期刊>Infection and immunity >Novel invasion determinant of enteropathogenic Escherichia coli plasmid pLV501 encodes the ability to invade intestinal epithelial cells and HEp-2 cells.
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Novel invasion determinant of enteropathogenic Escherichia coli plasmid pLV501 encodes the ability to invade intestinal epithelial cells and HEp-2 cells.

机译：肠致病性大肠杆菌质粒pLV501的新型入侵决定因子编码入侵肠上皮细胞和HEp-2细胞的能力。

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An Escherichia coli K-12 transformant carrying 96.5-kb plasmid pLV501 from enteropathogenic E. coli (EPEC) strain K798 is able to produce the same characteristic attaching-effacing lesions in a rabbit ileal biopsy explant model as its parent strain. Cloned EcoRI-SalI DNA restriction fragments from this plasmid failed to reproduce the attaching-effacing lesions, but one recombinant plasmid, pLV527, containing 4.5 kb of pLV501 DNA, conferred on E. coli DH1 transformants the ability to invade enterocytes in the rabbit explant model. DH1(pLV527) was also able to adhere to and invade HEp-2 cells. The relative invasive ability of DH1(pLV527) was quantified by recovery of internalized bacteria following gentamicin treatment of infected HEp-2 monolayers. DH1(pLV527) was 1,000-fold more invasive than DH1 carrying pBR322 or a recombinant plasmid which had no physiological effect on ileal biopsy explants but was less invasive than an enteroinvasive E. coli strain or a transformant carrying the cloned invasion genes of Shigella flexneri. Invasion by DH1(pLV501) could also be detected but occurred at a level 30 times lower than that by DH1(pLV527). Colony-hybridization of the pLV527 insert against a panel of 49 EPEC and related strains revealed that only 11 contained pLV527-hybridizing sequences; thus, the invasion determinant is not an essential component of the attachment-effacement pathogenic mechanism. One pLV527-hybridizing strain displayed both attachment-effacement and invasiveness in the rabbit ileal biopsy explant model. No significant hybridization was observed to non-EPEC invasive pathogenic enteric bacteria, indicating that the invasion determinant encoded on pLV527 is distinct from those used by these organisms.
机译：携带来自肠致病性大肠杆菌（EPEC）菌株K798的96.5-kb质粒pLV501的大肠杆菌K-12转化子能够在兔回肠活检外植体模型中产生与其亲代菌株相同的特征性附着表面损伤。从该质粒克隆的EcoRI-SalI DNA限制性片段未能重现附着的病灶，但是一个包含4.5 kb pLV501 DNA的重组质粒pLV527赋予了大肠杆菌DH1转化子侵袭兔外植体模型肠细胞的能力。 。 DH1（pLV527）也能够粘附和侵袭HEp-2细胞。 DH1（pLV527）的相对侵袭能力通过庆大霉素处理感染的HEp-2单层细菌后内在细菌的回收来定量。 DH1（pLV527）的侵害性比携带pBR322的DH1或对回肠活检外植体无生理影响但比携带肠炎志贺氏菌克隆的侵袭基因的转化株的侵害性弱的重组质粒的侵害力高1,000倍。 DH1（pLV501）的入侵也可以被检测到，但发生率比DH1（pLV527）低30倍。 pLV527插入片段针对49个EPEC和相关菌株的菌落杂交显示只有11个包含pLV527杂交序列。因此，入侵决定因素不是附着面致病机制的必要组成部分。一株pLV527杂交菌株在兔回肠活检外植体模型中表现出附着面和侵袭性。没有观察到与非EPEC侵袭性病原性肠道细菌的显着杂交，表明pLV527上编码的侵袭决定簇与这些生物体所使用的不同。

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