Enteropathogenic
            <i>Escherichia coli</i>
            Virulence Genes Encoding Secreted Signalling Proteins Are Essential for Modulation of Caco-2 Cell Electrolyte Transport

Collington, Georgina K.; Booth, I W; Donnenberg, Michael S.; Kaper, James B.; Knutton, Stuart

doi:10.1128/iai.66.12.6049-6053.1998

Cited by 38 publications

(14 citation statements)

References 35 publications

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“…Uninfected macrophages were used as control. To maximize LEE gene expression EPEC strains were primed for 3 h in Dulbecco's modified Eagle's medium (DMEM) prior to infection ( Collington et al ., 1998 ). In order to assist detection of bacteria in infected macrophages, strains were transformed with a GFP-expressing plasmid (pFVp25.1).…”

Section: Resultsmentioning

confidence: 99%

EspJ of enteropathogenic and enterohaemorrhagic Escherichia coli inhibits opsono-phagocytosis

et al. 2008

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SummaryA key strategy in microbial pathogenesis is the subversion of the first line of cellular immune defences presented by professional phagocytes. Enteropathogenic and enterohaemorrhagic Escherichia coli (EPEC and EHEC respectively) remain extracellular while colonizing the gut mucosa by attaching and effacing mechanism. EPEC use the type three secretion system effector protein EspF to prevent their own uptake into macrophages. EPEC can also block in trans the internalization of IgG-opsonized particles. In this study, we show that EspJ is the type three secretion system effector protein responsible for trans-inhibition of macrophage opsonophagocytosis by both EPEC and EHEC. While EspF plays no role in trans-inhibition of opsonophagocytosis, espJ mutants of EPEC or EHEC are unable to block uptake of opsonized sheep red blood cells (RBC), a phenotype that is rescued upon complementation with the espJ gene. Importantly, ectopic expression of EspJ EHEC in phagocytes is sufficient to inhibit internalization of both IgG-and C3bi-opsonized RBC. These results suggest that EspJ targets a basic mechanism common to these two unrelated phagocytic receptors. Moreover, EspF and EspJ target independent aspects of the phagocytic function of mammalian macrophages in vitro.

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Section: Resultsmentioning

confidence: 99%

EspJ of enteropathogenic and enterohaemorrhagic Escherichia coli inhibits opsono-phagocytosis

et al. 2008

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“…The examined salmonellae adhered to well-differentiated and to a lesser degree to nondifferentiated Caco-2 cells several times better than the lactobacilli ( Figure 1 ). Ours [ 23 ] and others [ 24 ] studies have shown that the colonization of lactobacilli takes place mainly in the intestinal crypts, whereas the pathogenic bacteria encounter mostly the upper part of intestinal villi and thus adhere to well-differentiated enterocytes (i.e., expressing the brush border). Among 5 LAB strains tested, the L. paracasei IBB2588 was the only one that preferred to attach to well-differentiated Caco-2 cells (this strain attached to well-differentiated cells 2.5-fold better than to nondifferentiated).…”

Section: Discussionmentioning

confidence: 99%

Competition of Lactobacillus paracasei with Salmonella enterica for Adhesion to Caco‐2 Cells

Jankowska

Laubitz

Antushevich

et al. 2008

BioMed Research International

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Competition of commensal and probiotic bacteria with pathogens for adhesion and colonization is one of the important protective mechanisms of gastrointestinal tract. In this study, we examined the ability of Lactobacillus paracasei to inhibit the adhesion of pathogenic Salmonella enterica to human colon adenocarcinoma Caco-2 cells. Caco-2 cells were grown for 6 or 21 days to obtain nondifferentiated or well-differentiated cells, respectively. In adhesion experiments, bacteria were added to the cells for 2 or 4 hours. The number of attached bacteria was expressed as colony-forming units (CFUs), Caco-2 cells were counted in hematocytometer. Both bacterial strains used adhered better to well-differentiated than to nondifferentiated Caco-2 cells, however, the amount of Salmonella adhered to Caco-2 after 2 hours of contact was 12-fold higher in comparison to L. paracasei and almost 27-fold higher after 4 hours of contact. Two types of experiments were done: coincubation (both bacteria were added to Caco-2 cells simultaneously), and preincubation (L. paracasei was incubated with Caco-2 cells first, and then S. enterica was added). In coincubation experiment, the presence of L. paracasei decreased S. enterica adhesion by 4-fold and in preincubation experiment even 7-fold. Generally, Lactobacillus spent culture supernatants (SCSs) acted weaker as inhibitors of Salmonella adhesion in comparison to the whole L. paracasei culture in coincubation experiment. In conclusion, the displacement of pathogens by lactic acid bacteria and its secretions showed here depends on the time of bacteria-epithelial cell contact, and also on the stage of Caco-2 differentiation.

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“…El aumento de la Isc depende del SSTT y el translocón, ya que cepas mutantes en EspA, EspB, EspD o EscN no aumentan la Isc. 57,58 Todos estos cambios en el gradiente electroquímico, que se exacerban por la disminución de la absorción intestinal y el incremento de la permeabilidad de la célula, son sin duda factores importantes dentro de la patogénesis de la diarrea secretora. 9,24 En un esfuerzo por conocer de modo más pormenorizado el mecanismo molecular de la diarrea, en fecha reciente los autores demostraron que EspC (una proteína autotransportadora secretada por EPEC mediante el sistema de secreción tipo V, SSTV) induce la salida masiva de iones al espacio extracelular.…”

Section: Figura 3 Detección De Epec Mediante El Ensayo De Adherenciaunclassified

Patogénesis molecular, epidemiología y diagnóstico de Escherichia coli enteropatógena

Vidal¹,

Canizalez-Román²,

Gutiérrez-Jiménez³

et al. 2007

Salud pública Méx

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Enteropathogenic Escherichia coli (EPEC) is a leading cause of diarrhea in infants less than two years of age in developing countries. To induce diarrhea EPEC uses several virulence factors acting on a still unknown and mysterious mechanism. The hallmark of EPEC infection is a histological intestinal alteration known as the attaching and effacing (A/E) lesion. The bacterium attaches intimately to the enterocyte and induces assembly of cytoskeleton intracellular actin on the cellular surface. Rearrangements of the actin cytoskeleton form a pedestal-like structure where bacterium tightly cups the cells, leading to degeneration of brush border microvilli. Although the mechanism of EPEC-induced pedestal formation has been dissected in detail, the overall mechanism of diarrhea is still obscure. It is believed that EPEC-mediated secretory diarrhea is related to a) intestinal microvilli effacement, b) massive loss of intracellular ions into the intestinal milieu and c) secretion of an EPEC enterotoxin. Epidemiological studies conducted in developing countries have shown that EPEC is one of the main bacteria frequently isolated from children with diarrhea, causing high morbidity and mortality rates. The microbiological diagnosis of EPEC-induced disease is performed with analytic methodologies different from those used by the standard microbiology laboratory, the most relevant being: a) serotypification, b) the adherence assay, c) FAS test, and d) the specific detection of virulence-involved genes (bfpA and eae genes) using molecular biology techniques. The purpose of this review is to update the most recent findings regarding the molecular pathogenesis of EPEC, its epidemiology in Mexico as well as other developing countries, and also the developed methodology for the diagnosis of EPEC infection.

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Enteropathogenic Escherichia coli Virulence Genes Encoding Secreted Signalling Proteins Are Essential for Modulation of Caco-2 Cell Electrolyte Transport

Cited by 38 publications

References 35 publications

EspJ of enteropathogenic and enterohaemorrhagic Escherichia coli inhibits opsono-phagocytosis

EspJ of enteropathogenic and enterohaemorrhagic Escherichia coli inhibits opsono-phagocytosis

Competition of Lactobacillus paracasei with Salmonella enterica for Adhesion to Caco‐2 Cells

Patogénesis molecular, epidemiología y diagnóstico de Escherichia coli enteropatógena

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