Escherichia coli isolates from inflammatory bowel diseases patients survive in macrophages and activate NLRP3 inflammasome

Marjorie De la Fuente; Luigi Franchi; Daniela Araya; David Díaz-Jiménez; Mauricio Olivares; Manuel Álvarez-Lobos; Douglas Golenbock; María Julieta González; Francisco López-Kostner; Rodrigo Quera; Gabriel Núñez; Roberto Vidal; Marcela A. Hermoso

doi:10.1016/j.ijmm.2014.01.002

Escherichia coli isolates from inflammatory bowel diseases patients survive in macrophages and activate NLRP3 inflammasome

Marjorie De la Fuente, Luigi Franchi, Daniela Araya, David Díaz-Jiménez, Mauricio Olivares, Manuel Álvarez-Lobos, Douglas Golenbock, María Julieta González, Francisco López-Kostner, Rodrigo Quera, Gabriel Núñez, Roberto Vidal, Marcela A. Hermoso^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

90 Scopus citations

Abstract

Crohn's disease (CD) is a multifactorial pathology associated with the presence of adherent-invasive Escherichia coli (AIEC) and NLRP3 polymorphic variants. The presence of intracellular E. coli in other intestinal pathologies (OIP) and the role of NLRP3-inflammasome in the immune response activated by these bacteria have not been investigated. In this study, we sought to characterize intracellular strains isolated from patients with CD, ulcerative colitis (UC) and OIP, and analyze NLRP3-inflammasome role in the immune response and bactericidal activity induced in macrophages exposed to invasive bacteria. For this, intracellular E. coli isolation from ileal biopsies, using gentamicin-protection assay, revealed a prevalence and CFU/biopsy of E. coli higher in biopsies from CD, UC and OIP patients than in controls. To characterize bacterial isolates, pulsed-field gel electrophoresis (PFGE) patterns, virulence genes, serogroup and phylogenetic group were analyzed. We found out that bacteria isolated from a given patient were closely related and shared virulence factors; however, strains from different patients were genetically heterogeneous. AIEC characteristics in isolated strains, such as invasive and replicative properties, were assessed in epithelial cells and macrophages, respectively. Some strains from CD and UC demonstrated AIEC properties, but not strains from OIP. Furthermore, the role of NLRP3 in pro-inflammatory cytokines production and bacterial elimination was determined in macrophages. E. coli strains induced IL-1β through NLRP3-dependent mechanism; however, their elimination by macrophages was independent of NLRP3. Invasiveness of intracellular E. coli strains into the intestinal mucosa and IL-1β production may contribute to CD and UC pathogenesis.

Original language	English
Pages (from-to)	384-392
Number of pages	9
Journal	International Journal of Medical Microbiology
Volume	304
Issue number	3-4
DOIs	https://doi.org/10.1016/j.ijmm.2014.01.002
State	Published - May 2014
Externally published	Yes

Bibliographical note

Funding Information:
Miguel ÓRyan for her helpful critical reading of the manuscript and Alfredo Torres for providing us the NRG857c sequenced AIEC strain. Funding support from FONDECYT 1120577 (MH), FONDECYT 1110260 (R.V.), PI2012-DA015 CLC, PI2010-DA CLC (R.Q.) , National Institutes of Health grants AI063331 and AI064748 (G.N.) and MECESUP UCH 0714 and CONICYT fellowships (M.D.L.F.).

Keywords

Crohn's disease
E. coli
IL-1β
Inflammasome
NLRP3

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/j.ijmm.2014.01.002

Cite this

De la Fuente, M., Franchi, L., Araya, D., Díaz-Jiménez, D., Olivares, M., Álvarez-Lobos, M., Golenbock, D., González, M. J., López-Kostner, F., Quera, R., Núñez, G., Vidal, R., & Hermoso, M. A. (2014). Escherichia coli isolates from inflammatory bowel diseases patients survive in macrophages and activate NLRP3 inflammasome. International Journal of Medical Microbiology, 304(3-4), 384-392. https://doi.org/10.1016/j.ijmm.2014.01.002

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title = "Escherichia coli isolates from inflammatory bowel diseases patients survive in macrophages and activate NLRP3 inflammasome",

abstract = "Crohn's disease (CD) is a multifactorial pathology associated with the presence of adherent-invasive Escherichia coli (AIEC) and NLRP3 polymorphic variants. The presence of intracellular E. coli in other intestinal pathologies (OIP) and the role of NLRP3-inflammasome in the immune response activated by these bacteria have not been investigated. In this study, we sought to characterize intracellular strains isolated from patients with CD, ulcerative colitis (UC) and OIP, and analyze NLRP3-inflammasome role in the immune response and bactericidal activity induced in macrophages exposed to invasive bacteria. For this, intracellular E. coli isolation from ileal biopsies, using gentamicin-protection assay, revealed a prevalence and CFU/biopsy of E. coli higher in biopsies from CD, UC and OIP patients than in controls. To characterize bacterial isolates, pulsed-field gel electrophoresis (PFGE) patterns, virulence genes, serogroup and phylogenetic group were analyzed. We found out that bacteria isolated from a given patient were closely related and shared virulence factors; however, strains from different patients were genetically heterogeneous. AIEC characteristics in isolated strains, such as invasive and replicative properties, were assessed in epithelial cells and macrophages, respectively. Some strains from CD and UC demonstrated AIEC properties, but not strains from OIP. Furthermore, the role of NLRP3 in pro-inflammatory cytokines production and bacterial elimination was determined in macrophages. E. coli strains induced IL-1β through NLRP3-dependent mechanism; however, their elimination by macrophages was independent of NLRP3. Invasiveness of intracellular E. coli strains into the intestinal mucosa and IL-1β production may contribute to CD and UC pathogenesis.",

keywords = "Crohn's disease, E. coli, IL-1β, Inflammasome, NLRP3",

author = "{De la Fuente}, Marjorie and Luigi Franchi and Daniela Araya and David D{\'i}az-Jim{\'e}nez and Mauricio Olivares and Manuel {\'A}lvarez-Lobos and Douglas Golenbock and Gonz{\'a}lez, {Mar{\'i}a Julieta} and Francisco L{\'o}pez-Kostner and Rodrigo Quera and Gabriel N{\'u}{\~n}ez and Roberto Vidal and Hermoso, {Marcela A.}",

note = "Funding Information: Miguel {\'O}Ryan for her helpful critical reading of the manuscript and Alfredo Torres for providing us the NRG857c sequenced AIEC strain. Funding support from FONDECYT 1120577 (MH), FONDECYT 1110260 (R.V.), PI2012-DA015 CLC, PI2010-DA CLC (R.Q.) , National Institutes of Health grants AI063331 and AI064748 (G.N.) and MECESUP UCH 0714 and CONICYT fellowships (M.D.L.F.).",

year = "2014",

month = may,

doi = "10.1016/j.ijmm.2014.01.002",

language = "English",

volume = "304",

pages = "384--392",

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De la Fuente, M, Franchi, L, Araya, D, Díaz-Jiménez, D, Olivares, M, Álvarez-Lobos, M, Golenbock, D, González, MJ, López-Kostner, F, Quera, R, Núñez, G, Vidal, R & Hermoso, MA 2014, 'Escherichia coli isolates from inflammatory bowel diseases patients survive in macrophages and activate NLRP3 inflammasome', International Journal of Medical Microbiology, vol. 304, no. 3-4, pp. 384-392. https://doi.org/10.1016/j.ijmm.2014.01.002

TY - JOUR

T1 - Escherichia coli isolates from inflammatory bowel diseases patients survive in macrophages and activate NLRP3 inflammasome

AU - De la Fuente, Marjorie

AU - Franchi, Luigi

AU - Araya, Daniela

AU - Díaz-Jiménez, David

AU - Olivares, Mauricio

AU - Álvarez-Lobos, Manuel

AU - Golenbock, Douglas

AU - González, María Julieta

AU - López-Kostner, Francisco

AU - Quera, Rodrigo

AU - Núñez, Gabriel

AU - Vidal, Roberto

AU - Hermoso, Marcela A.

N1 - Funding Information: Miguel ÓRyan for her helpful critical reading of the manuscript and Alfredo Torres for providing us the NRG857c sequenced AIEC strain. Funding support from FONDECYT 1120577 (MH), FONDECYT 1110260 (R.V.), PI2012-DA015 CLC, PI2010-DA CLC (R.Q.) , National Institutes of Health grants AI063331 and AI064748 (G.N.) and MECESUP UCH 0714 and CONICYT fellowships (M.D.L.F.).

PY - 2014/5

Y1 - 2014/5

N2 - Crohn's disease (CD) is a multifactorial pathology associated with the presence of adherent-invasive Escherichia coli (AIEC) and NLRP3 polymorphic variants. The presence of intracellular E. coli in other intestinal pathologies (OIP) and the role of NLRP3-inflammasome in the immune response activated by these bacteria have not been investigated. In this study, we sought to characterize intracellular strains isolated from patients with CD, ulcerative colitis (UC) and OIP, and analyze NLRP3-inflammasome role in the immune response and bactericidal activity induced in macrophages exposed to invasive bacteria. For this, intracellular E. coli isolation from ileal biopsies, using gentamicin-protection assay, revealed a prevalence and CFU/biopsy of E. coli higher in biopsies from CD, UC and OIP patients than in controls. To characterize bacterial isolates, pulsed-field gel electrophoresis (PFGE) patterns, virulence genes, serogroup and phylogenetic group were analyzed. We found out that bacteria isolated from a given patient were closely related and shared virulence factors; however, strains from different patients were genetically heterogeneous. AIEC characteristics in isolated strains, such as invasive and replicative properties, were assessed in epithelial cells and macrophages, respectively. Some strains from CD and UC demonstrated AIEC properties, but not strains from OIP. Furthermore, the role of NLRP3 in pro-inflammatory cytokines production and bacterial elimination was determined in macrophages. E. coli strains induced IL-1β through NLRP3-dependent mechanism; however, their elimination by macrophages was independent of NLRP3. Invasiveness of intracellular E. coli strains into the intestinal mucosa and IL-1β production may contribute to CD and UC pathogenesis.

AB - Crohn's disease (CD) is a multifactorial pathology associated with the presence of adherent-invasive Escherichia coli (AIEC) and NLRP3 polymorphic variants. The presence of intracellular E. coli in other intestinal pathologies (OIP) and the role of NLRP3-inflammasome in the immune response activated by these bacteria have not been investigated. In this study, we sought to characterize intracellular strains isolated from patients with CD, ulcerative colitis (UC) and OIP, and analyze NLRP3-inflammasome role in the immune response and bactericidal activity induced in macrophages exposed to invasive bacteria. For this, intracellular E. coli isolation from ileal biopsies, using gentamicin-protection assay, revealed a prevalence and CFU/biopsy of E. coli higher in biopsies from CD, UC and OIP patients than in controls. To characterize bacterial isolates, pulsed-field gel electrophoresis (PFGE) patterns, virulence genes, serogroup and phylogenetic group were analyzed. We found out that bacteria isolated from a given patient were closely related and shared virulence factors; however, strains from different patients were genetically heterogeneous. AIEC characteristics in isolated strains, such as invasive and replicative properties, were assessed in epithelial cells and macrophages, respectively. Some strains from CD and UC demonstrated AIEC properties, but not strains from OIP. Furthermore, the role of NLRP3 in pro-inflammatory cytokines production and bacterial elimination was determined in macrophages. E. coli strains induced IL-1β through NLRP3-dependent mechanism; however, their elimination by macrophages was independent of NLRP3. Invasiveness of intracellular E. coli strains into the intestinal mucosa and IL-1β production may contribute to CD and UC pathogenesis.

KW - Crohn's disease

KW - E. coli

KW - IL-1β

KW - Inflammasome

KW - NLRP3

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M3 - Article

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SN - 1438-4221

VL - 304

SP - 384

EP - 392

JO - International Journal of Medical Microbiology

JF - International Journal of Medical Microbiology

IS - 3-4

ER -

Escherichia coli isolates from inflammatory bowel diseases patients survive in macrophages and activate NLRP3 inflammasome

Abstract

Bibliographical note

Keywords

UN SDGs

Access to Document

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