Human Single-chain Variable Fragments Neutralize Pseudomonas aeruginosa Quorum Sensing Molecule, 3O-C12-HSL, and Prevent Cells From the HSL-mediated Apoptosis

Frontiers in Microbiology
Sirijan SantajitNitaya Indrawattana

Abstract

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 dock...Continue Reading

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Citations

Nov 3, 2020·Frontiers in Cellular and Infection Microbiology·Pengfei SheYong Wu
Jun 3, 2021·Vaccines·Jaslan DensumiteWanpen Chaicumpa

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Methods Mentioned

BETA
phage display
PCR
flow cytometry
Fluorescence
confocal microscopy
ELISA
electron microscopy

Software Mentioned

Discovery studio visualizer
CLICK
GraphPad Prism
PROCHECK
TASSER
Zeiss LSM Image Browser
Autodock Vina
FACSDiva
ModRefiner

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