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Human Single-chain Variable Fragments Neutralize Pseudomonas aeruginosa Quorum Sensing Molecule, 3O-C12-HSL, and Prevent Cells From the HSL-mediated Apoptosis
The quorum sensing (QS) signaling molecule, N-(3-oxododecanoyl)-L-homoserine lactone (3O-C12-HSL), contributes to the pathogenesis of Pseudomonas aeruginosa by regulating expression of the bacterial virulence factors that cause intense inflammation and toxicity in the infected host. As such, the QS...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Frontiers Media S.A.
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7326786/ https://www.ncbi.nlm.nih.gov/pubmed/32670218 http://dx.doi.org/10.3389/fmicb.2020.01172 |
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author | Santajit, Sirijan Seesuay, Watee Mahasongkram, Kodchakorn Sookrung, Nitat Pumirat, Pornpan Ampawong, Sumate Reamtong, Onrapak Chongsa-Nguan, Manas Chaicumpa, Wanpen Indrawattana, Nitaya |
author_facet | Santajit, Sirijan Seesuay, Watee Mahasongkram, Kodchakorn Sookrung, Nitat Pumirat, Pornpan Ampawong, Sumate Reamtong, Onrapak Chongsa-Nguan, Manas Chaicumpa, Wanpen Indrawattana, Nitaya |
author_sort | Santajit, Sirijan |
collection | PubMed |
description | The quorum sensing (QS) signaling molecule, N-(3-oxododecanoyl)-L-homoserine lactone (3O-C12-HSL), contributes to the pathogenesis of Pseudomonas aeruginosa by regulating expression of the bacterial virulence factors that cause intense inflammation and toxicity in the infected host. As such, the QS molecule is an attractive therapeutic target for direct-acting inhibitors. Several substances, both synthetic and naturally derived products, have shown effectiveness against detrimental 3O-C12-HSL activity. Unfortunately, these compounds are relatively toxic to mammalian cells, which limits their clinical application. In this study, fully human single-chain variable fragments (HuscFvs) that bind to P. aeruginosa haptenic 3O-C12-HSL were generated based on the principle of antibody polyspecificity and molecular mimicry of antigenic molecules. The HuscFvs neutralized 3O-C12-HSL activity and prevented mammalian cells from the HSL-mediated apoptosis, as observed by Annexin V/PI staining assay, sub-G1 arrest population investigation, transmission electron microscopy for ultrastructural morphology of mitochondria, and confocal microscopy for nuclear condensation and DNA fragmentation. Computerized homology modeling and intermolecular docking predicted that the effective HuscFvs interacted with several regions of the bacterially derived ligand that possibly conferred neutralizing activity. The effective HuscFvs should be tested further in vitro on P. aeruginosa phenotypes as well as in vivo as a sole or adjunctive therapeutic agent against P. aeruginosa infections, especially in antibiotic-resistant cases. |
format | Online Article Text |
id | pubmed-7326786 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-73267862020-07-14 Human Single-chain Variable Fragments Neutralize Pseudomonas aeruginosa Quorum Sensing Molecule, 3O-C12-HSL, and Prevent Cells From the HSL-mediated Apoptosis Santajit, Sirijan Seesuay, Watee Mahasongkram, Kodchakorn Sookrung, Nitat Pumirat, Pornpan Ampawong, Sumate Reamtong, Onrapak Chongsa-Nguan, Manas Chaicumpa, Wanpen Indrawattana, Nitaya Front Microbiol Microbiology The quorum sensing (QS) signaling molecule, N-(3-oxododecanoyl)-L-homoserine lactone (3O-C12-HSL), contributes to the pathogenesis of Pseudomonas aeruginosa by regulating expression of the bacterial virulence factors that cause intense inflammation and toxicity in the infected host. As such, the QS molecule is an attractive therapeutic target for direct-acting inhibitors. Several substances, both synthetic and naturally derived products, have shown effectiveness against detrimental 3O-C12-HSL activity. Unfortunately, these compounds are relatively toxic to mammalian cells, which limits their clinical application. In this study, fully human single-chain variable fragments (HuscFvs) that bind to P. aeruginosa haptenic 3O-C12-HSL were generated based on the principle of antibody polyspecificity and molecular mimicry of antigenic molecules. The HuscFvs neutralized 3O-C12-HSL activity and prevented mammalian cells from the HSL-mediated apoptosis, as observed by Annexin V/PI staining assay, sub-G1 arrest population investigation, transmission electron microscopy for ultrastructural morphology of mitochondria, and confocal microscopy for nuclear condensation and DNA fragmentation. Computerized homology modeling and intermolecular docking predicted that the effective HuscFvs interacted with several regions of the bacterially derived ligand that possibly conferred neutralizing activity. The effective HuscFvs should be tested further in vitro on P. aeruginosa phenotypes as well as in vivo as a sole or adjunctive therapeutic agent against P. aeruginosa infections, especially in antibiotic-resistant cases. Frontiers Media S.A. 2020-06-24 /pmc/articles/PMC7326786/ /pubmed/32670218 http://dx.doi.org/10.3389/fmicb.2020.01172 Text en Copyright © 2020 Santajit, Seesuay, Mahasongkram, Sookrung, Pumirat, Ampawong, Reamtong, Chongsa-Nguan, Chaicumpa and Indrawattana. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Microbiology Santajit, Sirijan Seesuay, Watee Mahasongkram, Kodchakorn Sookrung, Nitat Pumirat, Pornpan Ampawong, Sumate Reamtong, Onrapak Chongsa-Nguan, Manas Chaicumpa, Wanpen Indrawattana, Nitaya Human Single-chain Variable Fragments Neutralize Pseudomonas aeruginosa Quorum Sensing Molecule, 3O-C12-HSL, and Prevent Cells From the HSL-mediated Apoptosis |
title | Human Single-chain Variable Fragments Neutralize Pseudomonas aeruginosa Quorum Sensing Molecule, 3O-C12-HSL, and Prevent Cells From the HSL-mediated Apoptosis |
title_full | Human Single-chain Variable Fragments Neutralize Pseudomonas aeruginosa Quorum Sensing Molecule, 3O-C12-HSL, and Prevent Cells From the HSL-mediated Apoptosis |
title_fullStr | Human Single-chain Variable Fragments Neutralize Pseudomonas aeruginosa Quorum Sensing Molecule, 3O-C12-HSL, and Prevent Cells From the HSL-mediated Apoptosis |
title_full_unstemmed | Human Single-chain Variable Fragments Neutralize Pseudomonas aeruginosa Quorum Sensing Molecule, 3O-C12-HSL, and Prevent Cells From the HSL-mediated Apoptosis |
title_short | Human Single-chain Variable Fragments Neutralize Pseudomonas aeruginosa Quorum Sensing Molecule, 3O-C12-HSL, and Prevent Cells From the HSL-mediated Apoptosis |
title_sort | human single-chain variable fragments neutralize pseudomonas aeruginosa quorum sensing molecule, 3o-c12-hsl, and prevent cells from the hsl-mediated apoptosis |
topic | Microbiology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7326786/ https://www.ncbi.nlm.nih.gov/pubmed/32670218 http://dx.doi.org/10.3389/fmicb.2020.01172 |
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