title: Glycolysis and pyrimidine biosynthesis are required for replication of adherent-invasive Escherichia coli in macrophages.
abstract: Adherent-invasive Escherichia coli (AIEC) have been implicated in the aetiology of Crohn's disease (CD), a chronic inflammatory bowel condition. It has been proposed that AIEC-infected macrophages produce high levels of pro-inflammatory cytokines thus contributing to the inflammation observed in CD. AIEC can replicate in macrophages and we wanted to determine if bacterial replication was linked to the high level of cytokine production associated with AIEC-infected macrophages. Therefore, we undertook a genetic analysis of the metabolic requirements for AIEC replication in the macrophage and we show that AIEC replication in this niche is dependent on bacterial glycolysis. In addition, our analyses indicate that AIEC have access to a wide range of nutrients in the macrophage, although the levels of purines and pyrimidines do appear to be limiting. Finally, we show that the macrophage response to AIEC infection is indistinguishable from the response to the non-replicating glycolysis mutant (ΔpfkAB) and a non-pathogenic strain of E. coli, MG1655. Therefore, AIEC does not appear to subvert the normal macrophage response to E. coli during infection.
authors.affiliation: School of Microbiology, University College Cork, Cork, Ireland.|APC Microbiome Institute, University College Cork, Cork, Ireland.
authors.forename: Aoife P
authors.initials: AP
authors.lastname: Thompson
authors.affiliation: School of Microbiology, University College Cork, Cork, Ireland.|APC Microbiome Institute, University College Cork, Cork, Ireland.
authors.forename: Ian
authors.initials: I
authors.lastname: O'Neill
authors.affiliation: School of Microbiology, University College Cork, Cork, Ireland.|APC Microbiome Institute, University College Cork, Cork, Ireland.
authors.forename: Emma J
authors.initials: EJ
authors.lastname: Smith
authors.affiliation: School of Microbiology, University College Cork, Cork, Ireland.|APC Microbiome Institute, University College Cork, Cork, Ireland.
authors.forename: John
authors.initials: J
authors.lastname: Catchpole
authors.affiliation: APC Microbiome Institute, University College Cork, Cork, Ireland.
authors.forename: Ailis
authors.initials: A
authors.lastname: Fagan
authors.affiliation: Glasgow Polyomics, University of Glasgow, Switchback Road, Glasgow G61 1QH, UK.
authors.forename: Karl E V
authors.initials: KEV
authors.lastname: Burgess
authors.affiliation: APC Microbiome Institute, University College Cork, Cork, Ireland.
authors.forename: Ruaidhri J
authors.initials: RJ
authors.lastname: Carmody
authors.affiliation: APC Microbiome Institute, University College Cork, Cork, Ireland.|School of Microbiology, University College Cork, Cork, Ireland.
authors.forename: David J
authors.initials: DJ
authors.lastname: Clarke
mesh_terms: D000818:Animals; D002460:Cell Line; D003424:Crohn Disease / Q000382:microbiology; D016207:Cytokines / Q000096:biosynthesis*; D004251:DNA Transposable Elements / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism*; D015723:Gene Library; D006019:Glycolysis / Q000235:genetics*; D006801:Humans; D008264:Macrophages / Q000382:microbiology*; D055432:Metabolomics; D051379:Mice; D011743:Pyrimidines / Q000096:biosynthesis*
chemical_list: D016207:Cytokines; D004251:DNA Transposable Elements; D011743:Pyrimidines