caa a22 4500 606176012 CHVBK 20210128100745.0 cr unu---uuuuu 210128e20150201xx s 000 0 eng 10.1007/s00018-014-1694-0 doi (NATIONALLICENCE)springer-10.1007/s00018-014-1694-0 Esculentin-1a(1-21)NH2: a frog skin-derived peptide for microbial keratitis [Elektronische Daten] [Satya Kolar, Vincenzo Luca, Hasna Baidouri, Giuseppe Mannino, Alison McDermott, Maria Mangoni] 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 16μM against reference strains or drug-resistant clinical isolates of P. aeruginosa without showing toxicity to human corneal epithelial cells up to 50μM. At 1μM the peptide rapidly killed bacterial cells and this activity was fully retained in 150mM sodium chloride and 70% (v/v) human basal tears, particularly against the virulent ATCC 19660 strain. Furthermore, its dropwise administration at 40μM to the ocular surface in a murine model of P. aeruginosa keratitis (three times daily, for 5days 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. Springer Basel, 2014 Innate immunity nationallicence Ocular surface infections nationallicence Antibiotic resistance nationallicence Amphibian skin antimicrobial peptides nationallicence AMP : Antimicrobial peptide nationallicence BAC : Benzalkonium chloride nationallicence CFU : Colony-forming units nationallicence Esc-1a(1-21)NH2 : Esculentin-1a(1-21)NH2 nationallicence hTCEpi : Telomerase-immortalized human cornel epithelial cells nationallicence LB : Luria-Bertani nationallicence MH : Mueller-Hinton nationallicence MIC : Minimal inhibitory concentration nationallicence MTT : 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide nationallicence MPO : Myeloperoxidase nationallicence PB : Phosphate buffer nationallicence PBS : Phosphate buffered saline nationallicence Kolar Satya College of Optometry, The Ocular Surface Institute, University of Houston, Houston, TX, USA aut Luca Vincenzo Department of Biochemical Sciences "A. Rossi Fanelli”, Istituto Pasteur-Fondazione Cenci Bolognetti, Sapienza University of Rome, Via degli Apuli 9, 00185, Rome, Italy aut Baidouri Hasna College of Optometry, The Ocular Surface Institute, University of Houston, Houston, TX, USA aut Mannino Giuseppe Ophthalmology Unit, NESMOS Department, S. Andrea Hospital, Faculty of Medicine and Psychology, Sapienza University of Rome, Rome, Italy aut McDermott Alison College of Optometry, The Ocular Surface Institute, University of Houston, Houston, TX, USA aut Mangoni Maria Department of Biochemical Sciences "A. Rossi Fanelli”, Istituto Pasteur-Fondazione Cenci Bolognetti, Sapienza University of Rome, Via degli Apuli 9, 00185, Rome, Italy aut Cellular and Molecular Life Sciences Springer Basel 72/3(2015-02-01), 617-627 1420-682X 72:3<617 2015 72 18 https://doi.org/10.1007/s00018-014-1694-0 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00018-014-1694-0 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Kolar Satya College of Optometry, The Ocular Surface Institute, University of Houston, Houston, TX, USA aut NATIONALLICENCE 700 1- Luca Vincenzo Department of Biochemical Sciences "A. Rossi Fanelli”, Istituto Pasteur-Fondazione Cenci Bolognetti, Sapienza University of Rome, Via degli Apuli 9, 00185, Rome, Italy aut NATIONALLICENCE 700 1- Baidouri Hasna College of Optometry, The Ocular Surface Institute, University of Houston, Houston, TX, USA aut NATIONALLICENCE 700 1- Mannino Giuseppe Ophthalmology Unit, NESMOS Department, S. Andrea Hospital, Faculty of Medicine and Psychology, Sapienza University of Rome, Rome, Italy aut NATIONALLICENCE 700 1- McDermott Alison College of Optometry, The Ocular Surface Institute, University of Houston, Houston, TX, USA aut NATIONALLICENCE 700 1- Mangoni Maria Department of Biochemical Sciences "A. Rossi Fanelli”, Istituto Pasteur-Fondazione Cenci Bolognetti, Sapienza University of Rome, Via degli Apuli 9, 00185, Rome, Italy aut NATIONALLICENCE 773 0- Cellular and Molecular Life Sciences Springer Basel 72/3(2015-02-01), 617-627 1420-682X 72:3<617 2015 72 18