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Replication of Crohn’s Disease Mucosal E. coli Isolates inside Macrophages Correlates with Resistance to Superoxide and Is Dependent on Macrophage NF-kappa B Activation

Identifieur interne : 000471 ( Main/Exploration ); précédent : 000470; suivant : 000472

Replication of Crohn’s Disease Mucosal E. coli Isolates inside Macrophages Correlates with Resistance to Superoxide and Is Dependent on Macrophage NF-kappa B Activation

Auteurs : Ahmed Tawfik [Irlande (pays)] ; Paul Knight [Royaume-Uni] ; Carrie A. Duckworth ; D. Mark Pritchard ; Jonathan M. Rhodes ; Barry J. Campbell

Source :

RBID : PMC:6630736

Abstract

Mucosa-associated Escherichia coli are increased in Crohn’s disease (CD) and colorectal cancer (CRC). CD isolates replicate within macrophages but the specificity of this effect for CD and its mechanism are unclear. Gentamicin exclusion assay was used to assess E. coli replication within J774.A1 murine macrophages. E. coli growth was assessed following acid, low-nutrient, nitrosative, oxidative and superoxide stress, mimicking the phagolysosome. Twelve of 16 CD E. coli isolates replicated >2-fold within J774.A1 macrophages; likewise for isolates from 6/7 urinary tract infection (UTI), 8/9 from healthy subjects, compared with 2/6 ulcerative colitis, 2/7 colorectal cancer and 0/3 laboratory strains. CD mucosal E. coli were tolerant of acidic, low-nutrient, nitrosative and oxidative stress. Replication within macrophages correlated strongly with tolerance to superoxide stress (rho = 0.44, p = 0.0009). Exemplar CD E. coli HM605 and LF82 were unable to survive within Nfκb1-/- murine bone marrow-derived macrophages. In keeping with this, pre-incubation of macrophages with hydrocortisone (0.6 µM for 24 h) caused 70.49 ± 12.11% inhibition of intra-macrophage replication. Thus, CD mucosal E. coli commonly replicate inside macrophages, but so do some UTI and healthy subject strains. Replication correlates with resistance to superoxide and is highly dependent on macrophage NF-κB signalling. This may therefore be a good therapeutic target.


Url:
DOI: 10.3390/pathogens8020074
PubMed: 31181736
PubMed Central: 6630736


Affiliations:


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Le document en format XML

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<title xml:lang="en" level="a" type="main">Replication of Crohn’s Disease Mucosal
<italic>E. coli</italic>
Isolates inside Macrophages Correlates with Resistance to Superoxide and Is Dependent on Macrophage NF-kappa B Activation</title>
<author>
<name sortKey="Tawfik, Ahmed" sort="Tawfik, Ahmed" uniqKey="Tawfik A" first="Ahmed" last="Tawfik">Ahmed Tawfik</name>
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<nlm:aff id="af1-pathogens-08-00074">Gastroenterology Research Unit, Department of Cellular & Molecular Physiology, Institute of Translational Medicine, University of Liverpool, Liverpool L69 3GE, UK;
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(A.T.);
<email>paul.knight@mft.nhs.uk</email>
(P.K.);
<email>carried@liv.ac.uk</email>
(C.A.D.);
<email>dmpritch@liv.ac.uk</email>
(D.M.P.);
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<name sortKey="Knight, Paul" sort="Knight, Paul" uniqKey="Knight P" first="Paul" last="Knight">Paul Knight</name>
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<email>ahmedtawfik13@yahoo.com</email>
(A.T.);
<email>paul.knight@mft.nhs.uk</email>
(P.K.);
<email>carried@liv.ac.uk</email>
(C.A.D.);
<email>dmpritch@liv.ac.uk</email>
(D.M.P.);
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<name sortKey="Duckworth, Carrie A" sort="Duckworth, Carrie A" uniqKey="Duckworth C" first="Carrie A." last="Duckworth">Carrie A. Duckworth</name>
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<nlm:aff id="af1-pathogens-08-00074">Gastroenterology Research Unit, Department of Cellular & Molecular Physiology, Institute of Translational Medicine, University of Liverpool, Liverpool L69 3GE, UK;
<email>ahmedtawfik13@yahoo.com</email>
(A.T.);
<email>paul.knight@mft.nhs.uk</email>
(P.K.);
<email>carried@liv.ac.uk</email>
(C.A.D.);
<email>dmpritch@liv.ac.uk</email>
(D.M.P.);
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<name sortKey="Pritchard, D Mark" sort="Pritchard, D Mark" uniqKey="Pritchard D" first="D. Mark" last="Pritchard">D. Mark Pritchard</name>
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<nlm:aff id="af1-pathogens-08-00074">Gastroenterology Research Unit, Department of Cellular & Molecular Physiology, Institute of Translational Medicine, University of Liverpool, Liverpool L69 3GE, UK;
<email>ahmedtawfik13@yahoo.com</email>
(A.T.);
<email>paul.knight@mft.nhs.uk</email>
(P.K.);
<email>carried@liv.ac.uk</email>
(C.A.D.);
<email>dmpritch@liv.ac.uk</email>
(D.M.P.);
<email>rhodesjm@liv.ac.uk</email>
(J.M.R.)</nlm:aff>
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<name sortKey="Rhodes, Jonathan M" sort="Rhodes, Jonathan M" uniqKey="Rhodes J" first="Jonathan M." last="Rhodes">Jonathan M. Rhodes</name>
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<nlm:aff id="af1-pathogens-08-00074">Gastroenterology Research Unit, Department of Cellular & Molecular Physiology, Institute of Translational Medicine, University of Liverpool, Liverpool L69 3GE, UK;
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<email>paul.knight@mft.nhs.uk</email>
(P.K.);
<email>carried@liv.ac.uk</email>
(C.A.D.);
<email>dmpritch@liv.ac.uk</email>
(D.M.P.);
<email>rhodesjm@liv.ac.uk</email>
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<name sortKey="Campbell, Barry J" sort="Campbell, Barry J" uniqKey="Campbell B" first="Barry J." last="Campbell">Barry J. Campbell</name>
<affiliation>
<nlm:aff id="af1-pathogens-08-00074">Gastroenterology Research Unit, Department of Cellular & Molecular Physiology, Institute of Translational Medicine, University of Liverpool, Liverpool L69 3GE, UK;
<email>ahmedtawfik13@yahoo.com</email>
(A.T.);
<email>paul.knight@mft.nhs.uk</email>
(P.K.);
<email>carried@liv.ac.uk</email>
(C.A.D.);
<email>dmpritch@liv.ac.uk</email>
(D.M.P.);
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<div type="abstract" xml:lang="en">
<p>Mucosa-associated
<italic>Escherichia coli</italic>
are increased in Crohn’s disease (CD) and colorectal cancer (CRC). CD isolates replicate within macrophages but the specificity of this effect for CD and its mechanism are unclear. Gentamicin exclusion assay was used to assess
<italic>E. coli</italic>
replication within J774.A1 murine macrophages.
<italic>E. coli</italic>
growth was assessed following acid, low-nutrient, nitrosative, oxidative and superoxide stress, mimicking the phagolysosome. Twelve of 16 CD
<italic>E. coli</italic>
isolates replicated >2-fold within J774.A1 macrophages; likewise for isolates from 6/7 urinary tract infection (UTI), 8/9 from healthy subjects, compared with 2/6 ulcerative colitis, 2/7 colorectal cancer and 0/3 laboratory strains. CD mucosal
<italic>E. coli</italic>
were tolerant of acidic, low-nutrient, nitrosative and oxidative stress. Replication within macrophages correlated strongly with tolerance to superoxide stress (rho = 0.44,
<italic>p</italic>
= 0.0009). Exemplar CD
<italic>E. coli</italic>
HM605 and LF82 were unable to survive within
<italic>Nfκb1
<sup>-/-</sup>
</italic>
murine bone marrow-derived macrophages. In keeping with this, pre-incubation of macrophages with hydrocortisone (0.6 µM for 24 h) caused 70.49 ± 12.11% inhibition of intra-macrophage replication. Thus, CD mucosal
<italic>E. coli</italic>
commonly replicate inside macrophages, but so do some UTI and healthy subject strains. Replication correlates with resistance to superoxide and is highly dependent on macrophage NF-κB signalling. This may therefore be a good therapeutic target.</p>
</div>
</front>
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Data generation: Wed Mar 25 22:43:59 2020. Site generation: Sun Jan 31 12:44:45 2021