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Bowman Club 2022, 25 March 2022
Poster abstract presentation
P-18 Impact of fluoroquinolones and aminoglycosides on P. aeruginosa virulence factor production and cytotoxicity
  1. Stephen Kaye1,
  2. Keri McLean1,
  3. Daniel M Foulkes1,2,
  4. Marta Sloniecka2,
  5. Dominic Byrne2,
  6. Atikah S Haneef1,
  7. Craig Winstanley3,
  8. Neil G Berry4,
  9. David G Fernig2
  1. 1Department of Eye and Vision Science, Institute of Ageing and Chronic Disease, University of Liverpool, UK
  2. 2Department of Biochemistry, Institute of Integrative Biology, University of Liverpool, UK
  3. 3Department of Clinical Infection, Institute of Infection and Global Health, University of Liverpool, UK
  4. 4Department of Chemistry, University of Liverpool, UK

Abstract

*Correspondence – Stephen Kaye: S.B.Kaye@liverpool.ac.uk

Introduction Pseudomonas aeruginosa injects toxins, ExoS or ExoU, into host cells via the type III secretion system (T3SS) which destroy cells and help evade the immune system. First-line fluoroquinolones demonstrate better in vitro activity against P. aeruginosa but in certain clinical situations aminoglycosides are more effective. We evaluate the effects of fluoroquinolones (moxifloxacin and ciprofloxacin) and aminoglycosides (tobramycin and gentamycin) on T3SS and toxin expression, and the associated toxicity in corneal epithelial cell infection models.

Methods Expression levels of pcrV (T3SS needle component) from ExoU-expressing PA103 and ExoS-expressing PA76026 after 16h incubation in each antimicrobial was detected using western blotting. qRT PCR detected mRNA levels of ExoU, ExoS, pcrV and ExsA (T3SS activating factor) after PA103 and PA76026 were exposed to tobramycin and moxifloxacin. LIVE/DEAD and LDH assays after 24h evaluated how the antimicrobials influenced acute cytotoxicity in a HCE-T cell scratch and infection model.

Results Tobramycin significantly reduced pcrV in both strains by 50.5–74.0% compared to the fluoroquinolones (p=0.001 and 0.003), even at low concentrations. Fluoroquinolones significantly increased pcrV by 57.0–81.8% (p=0.004 and 0.003). mRNA levels of ExoU, ExoS, pcrV and ExsA were reduced by tobramycin but moxifloxacin increased pcrV, ExsA and ExoS. Tobramycin, despite more bacterial expansion compared to the same relative concentrations of fluoroquinolones, reduced ExoU/ExoS cytotoxicity and allowed complete wound healing.

Discussion Tobramycin downregulates T3SS expression and reduces ExoS/ExoU mediated cytotoxicity which protects infected HCE-T cells even at low concentrations. Fluoroquinolones however upregulated T3SS and do not negate the cytotoxic effects.

http://creativecommons.org/licenses/by-nc/4.0/

This is an open access article distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited, appropriate credit is given, any changes made indicated, and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/ .

http://dx.doi.org/10.1136/bmjophth-2022-BCM.15

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