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Targeted amplification of the oxidative burst in neutrophils as a bactericidal strategy against uropathogenic Escherichia coli / LOUIE BARNES

Swansea University Author: LOUIE BARNES

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Abstract

Urinary tract infections (UTIs), commonly caused by the Gram-negative bacteria Escherichia. coli, affect 50% of women and 12% of men in their lifetimes and represent an enormous burden on healthcare providers. UTIs have a high rate of recurrence and a low rate of resolution without antibiotic interv...

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Published: Swansea, Wales, UK 2025
Institution: Swansea University
Degree level: Master of Research
Degree name: MSc by Research
Supervisor: Chick, Heather ; Wilkinson, Tom
URI: https://cronfa.swan.ac.uk/Record/cronfa69857
first_indexed 2025-06-30T15:12:37Z
last_indexed 2025-07-01T05:28:40Z
id cronfa69857
recordtype RisThesis
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spelling 2025-06-30T16:28:05.0618805 v2 69857 2025-06-30 Targeted amplification of the oxidative burst in neutrophils as a bactericidal strategy against uropathogenic Escherichia coli 1a0e16f9f38675af1dd39f35c36a76fd LOUIE BARNES LOUIE BARNES true false 2025-06-30 Urinary tract infections (UTIs), commonly caused by the Gram-negative bacteria Escherichia. coli, affect 50% of women and 12% of men in their lifetimes and represent an enormous burden on healthcare providers. UTIs have a high rate of recurrence and a low rate of resolution without antibiotic intervention, making them a driving force in the rise of antimicrobial resistance (AMR). Alternative therapies such as immunomodulation could aid in reduction of antibiotic prescription and AMR. Small molecule compounds RE-04-001 and RE-04-006 selectively target Formyl peptide receptor 1 (FPR1) in neutrophils, the most abundant innate immune cells in UTIs, resulting in the generation of toxic reactive oxygen species via the oxidative burst. This research aimed to characterize interactions between FPR1-stimulated neutrophils and Uropathogenic E. coli (UPEC). Neutrophils isolated from whole blood were co-cultured with UPEC strain CFT-073 or laboratory strain K12 and co-treated with FPR1-stimulating RE-04-001, RE-04-006 or N-Formylmethionyl-leucyl-phenylalanine. The internal killing ability of neutrophils was assessed using a gentamicin protection assay. Light microscopy was used to investigate phagocytosis. Neutrophil morphology following FPR1 stimulation was observed using flow cytometry, and viability was tested with DRAQ7™. Finally, confocal microscopy assessed FPR1-driven NETosis. FPR1 stimulation was found to enhance intracellular killing of CFT-073 but not K12. Extracellular killing of K12 but not UPEC CFT-073 in the supernatant was enhanced by FPR1 activation. FPR1 stimulation was found to have no significant effects on the phagocytosis of E. coli, contrasting other findings in the literature. FPR1-stimulating compounds had no impact on neutrophil viability but caused a significant shift in their morphology. Finally, FPR1 stimulation was observed to induce NETosis in neutrophils after 3 hours but no NETosis was recorded in co-cultures with E. coli. This work highlights FPR1 as a target for immunostimulatory therapy which is a critical alternative to or co-therapy with antibiotic treatment in UTIs. E-Thesis Swansea, Wales, UK Urinary tract infection, UPEC, Escherichia coli, Neutrophils, Reactive oxygen species, Formyl peptide receptor 1, Oxidative stress 27 6 2025 2025-06-27 ORCiD identifier: https://orcid.org/0009-0006-5519-8845 COLLEGE NANME COLLEGE CODE Swansea University Chick, Heather ; Wilkinson, Tom Master of Research MSc by Research Neutrocure Neutrocure 2025-06-30T16:28:05.0618805 2025-06-30T16:03:54.4794416 Faculty of Medicine, Health and Life Sciences Swansea University Medical School - Biomedical Science LOUIE BARNES 1 69857__34634__8abcc1abd8e9418f97667e78afe73f64.pdf Barnes_Louie_A_MSc_Research_Thesis_Final_Cronfa.pdf 2025-06-30T16:21:26.8864996 Output 2610777 application/pdf E-Thesis – open access true Copyright: The Author, Louie Arron Barnes, 2025. Licensed under a Creative Commons Attribution 4.0 International (CC BY 4.0) License. Third party content is excluded for use under the license terms. true eng http://creativecommons.org/licenses/by/4.0/
title Targeted amplification of the oxidative burst in neutrophils as a bactericidal strategy against uropathogenic Escherichia coli
spellingShingle Targeted amplification of the oxidative burst in neutrophils as a bactericidal strategy against uropathogenic Escherichia coli
LOUIE BARNES
title_short Targeted amplification of the oxidative burst in neutrophils as a bactericidal strategy against uropathogenic Escherichia coli
title_full Targeted amplification of the oxidative burst in neutrophils as a bactericidal strategy against uropathogenic Escherichia coli
title_fullStr Targeted amplification of the oxidative burst in neutrophils as a bactericidal strategy against uropathogenic Escherichia coli
title_full_unstemmed Targeted amplification of the oxidative burst in neutrophils as a bactericidal strategy against uropathogenic Escherichia coli
title_sort Targeted amplification of the oxidative burst in neutrophils as a bactericidal strategy against uropathogenic Escherichia coli
author_id_str_mv 1a0e16f9f38675af1dd39f35c36a76fd
author_id_fullname_str_mv 1a0e16f9f38675af1dd39f35c36a76fd_***_LOUIE BARNES
author LOUIE BARNES
author2 LOUIE BARNES
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publishDate 2025
institution Swansea University
college_str Faculty of Medicine, Health and Life Sciences
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hierarchy_top_id facultyofmedicinehealthandlifesciences
hierarchy_top_title Faculty of Medicine, Health and Life Sciences
hierarchy_parent_id facultyofmedicinehealthandlifesciences
hierarchy_parent_title Faculty of Medicine, Health and Life Sciences
department_str Swansea University Medical School - Biomedical Science{{{_:::_}}}Faculty of Medicine, Health and Life Sciences{{{_:::_}}}Swansea University Medical School - Biomedical Science
document_store_str 1
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description Urinary tract infections (UTIs), commonly caused by the Gram-negative bacteria Escherichia. coli, affect 50% of women and 12% of men in their lifetimes and represent an enormous burden on healthcare providers. UTIs have a high rate of recurrence and a low rate of resolution without antibiotic intervention, making them a driving force in the rise of antimicrobial resistance (AMR). Alternative therapies such as immunomodulation could aid in reduction of antibiotic prescription and AMR. Small molecule compounds RE-04-001 and RE-04-006 selectively target Formyl peptide receptor 1 (FPR1) in neutrophils, the most abundant innate immune cells in UTIs, resulting in the generation of toxic reactive oxygen species via the oxidative burst. This research aimed to characterize interactions between FPR1-stimulated neutrophils and Uropathogenic E. coli (UPEC). Neutrophils isolated from whole blood were co-cultured with UPEC strain CFT-073 or laboratory strain K12 and co-treated with FPR1-stimulating RE-04-001, RE-04-006 or N-Formylmethionyl-leucyl-phenylalanine. The internal killing ability of neutrophils was assessed using a gentamicin protection assay. Light microscopy was used to investigate phagocytosis. Neutrophil morphology following FPR1 stimulation was observed using flow cytometry, and viability was tested with DRAQ7™. Finally, confocal microscopy assessed FPR1-driven NETosis. FPR1 stimulation was found to enhance intracellular killing of CFT-073 but not K12. Extracellular killing of K12 but not UPEC CFT-073 in the supernatant was enhanced by FPR1 activation. FPR1 stimulation was found to have no significant effects on the phagocytosis of E. coli, contrasting other findings in the literature. FPR1-stimulating compounds had no impact on neutrophil viability but caused a significant shift in their morphology. Finally, FPR1 stimulation was observed to induce NETosis in neutrophils after 3 hours but no NETosis was recorded in co-cultures with E. coli. This work highlights FPR1 as a target for immunostimulatory therapy which is a critical alternative to or co-therapy with antibiotic treatment in UTIs.
published_date 2025-06-27T06:48:04Z
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