Esculentin-1a(1-21)NH2: a frog skin-derived peptide for microbial keratitis.

Abstract

Pseudomonas aeruginosa is the primary bacterial pathogen causing contact lens related keratitis. Available ophthalmic agents have reduced efficacy and antimicrobial peptides (AMPs) hold promise as future antibiotics. Here we investigated the in vitro and in vivo anti-Pseudomonal activity of esculentin-1a(1-21)NH2, derived from a frog skin AMP. The data revealed a minimum inhibitory concentration between 2 and 16 μM against reference strains or drug-resistant clinical isolates of P. aeruginosa without showing toxicity to human corneal epithelial cells up to 50 μM. At 1 μM the peptide rapidly killed bacterial cells and this activity was fully retained in 150 mM sodium chloride and 70 % (v/v) human basal tears, particularly against the virulent ATCC 19660 strain. Furthermore, its dropwise administration at 40 μM to the ocular surface in a murine model of P. aeruginosa keratitis (three times daily, for 5 days post-infection) resulted in a significant reduction of infection. The mean clinical score was 2.89 ± 0.26 compared to 3.92 ± 0.08 for the vehicle control. In addition, the corneal level of viable bacteria in the peptide treated animals was significantly lower with a difference of 4 log10 colony counts, compared to 7.7 log10 cells recovered in the control. In parallel, recruitment of inflammatory cells was reduced by half compared to that found in the untreated eyes. Similar results were obtained when esculentin-1a(1-21)NH2 was applied prior to induction of keratitis. Overall, our findings highlight esculentin-1a(1-21)NH2 as an attractive candidate for the development of novel topical pharmaceuticals against Pseudomonas keratitis.

• Authors: Satya Sree N Kolar, Vincenzo Luca, Hasna Baidouri, Giuseppe Mannino, Alison M McDermott, Maria Luisa Mangoni
• Journal: Cellular and molecular life sciences : CMLS (Cell Mol Life Sci) Vol. 72 Issue 3 Pg. 617-627 (Feb 2015) ISSN: 1420-9071 [Electronic] Switzerland
• PMID: 25086859 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)

Chemical References

• Amphibian Proteins
• Antimicrobial Cationic Peptides
• Tetrazolium Salts
• Thiazoles
• esculentin protein, Rana esculenta
• thiazolyl blue

Topics

• Amphibian Proteins (isolation & purification, pharmacology)
• Analysis of Variance
• Animals
• Antimicrobial Cationic Peptides (isolation & purification, pharmacology)
• Anura (metabolism)
• Contact Lenses (adverse effects, microbiology)
• Cornea (cytology)
• Epithelial Cells (drug effects)
• Humans
• Keratitis (drug therapy, etiology, microbiology)
• Pseudomonas aeruginosa (drug effects)
• Skin (chemistry)
• Tetrazolium Salts
• Thiazoles