Antimicrobial and Immunomodulatory Activities of PR-39 Derived Peptides

Veldhuizen, Edwin J.A.; Schneider, Viktoria A. F.; Agustiandari, Herfita; Dijk, Albert van; Bokhoven, Johanna L.M. Tjeerdsma-van; Bikker, Floris J.; Haagsman, Henk P.

doi:10.1371/journal.pone.0095939

Cited by 103 publications

(80 citation statements)

References 39 publications

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“…In the present study more than 70% ATP was released by melimine and approximately 35% by Mel4 following 2 minutes exposure, and the amount of ATP released plateaued at after four minutes exposure to both peptides. The effect of melimine is similar to the AMP porcine cathelicidin’s PR-39 effect of E. coli which induced 80% ATP leakage (58). The leakage of ATP results in depletion of intracellular ATP which in turn can lead to bacterial death (56).…”

Section: Discussionmentioning

confidence: 74%

“…For determination of internal ATP the bacterial pellets were resuspended in boiling 100 mM Tris, 4 mM EDTA pH (7.4) and further incubated for 2 mins at 100 °C to lyse all the cells. The lysed cells were centrifuged at 9600 × g for two minutes and the supernatant was kept on ice until further analysis (41). Subsequently, both total and extracellular ATP were determined using an ATP bioluminescence kit (Invitrogen, Eugene, Oregon, USA) according to manufacturer’s instructions.…”

Section: Methodsmentioning

confidence: 99%

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Comparative mode of action of antimicrobial peptide melimine and its derivative Mel4 against Pseudomonas aeruginosa

Yasir

Dutta

Willcox

2018

Preprint

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Society for Microbiology annual meetings in 2017. There are no conflicts of interest for any 31 of the authors that could have influenced the results of this work. Prof. Mark Willcox holds 32 the patent for the melimine peptide. 33 2 ABSTRACT 34Melimine and Mel4 are chimeric cationic peptides with broad spectrum antimicrobial 35 activity, and recent investigations have shown that they are highly biocompatible with animal 36 model and human clinical trials. The current study examined the mechanism of action of 37 these two antimicrobial peptides against P. aeruginosa with a series of investigations. 38 Antimicrobial activities were determined by MIC and MBC. Endotoxin neutralization was 39 determined using the LAL assay, effect on the cytoplasmic membrane was evaluated using 40 DiSC(3)-5 and Sytox green stains, and Syto-9 and PI dyes using flow cytometry. Release of 41 cytoplasmic materials (ATP and DNA/RNA) were determined using ATP luminescence and 42 increase in OD 260nm . The ability to lyse bacteria was studied by measuring a decrease in 43 OD 620nm . The MIC of the peptides remained low against P. aeruginosa strains, which showed 44 efficient neutralization of LPS, indicating their role in the anti-pseudomonas and LPS binding 45 activities. Both AMPs rapidly (starting at 30 seconds) depolarized P. aeruginosa cytoplasmic 46 membrane leading to reduction in viability. Melimine was responsible for more ATP release 47 (75%) compared to Mel4 (36%) (P<0.001) following two minutes exposure. For both 48 peptides, Sytox green entered cells after five minutes of incubation. Flow cytometry 49 demonstrated that both the AMPs permeabilized the cell membrane at 30 minutes and 50 followed by increasing permeability. Similar results were found with DNA/RNA release 51 experiments. Overall, melimine showed higher ability of membrane disruption, cell lysis 52 compared to Mel4 (P<0.001). Knowledge regarding mechanism of action of these two AMPs 53 would be helpful in making them as anti-pseudomonas drug. 54KEYWORDS P. aeruginosa, Antimicrobial peptides, mechanism of action, membrane 55 disruption 56 57 3 58 59 Pseudomonas aeruginosa is a metabolically versatile ubiquitous Gram-negative opportunistic 60 pathogen that can cause infections in animals and plants (1). P. aeruginosa is responsible for 61 localized to systemic infections in humans, which can be life threatening. Over the years, P. 62 aeruginosa has become one of the most frequent causative agents of nosocomial infections, 63 associated with substantial morbidity and mortality (2). The current standards of care to treat 64 P. aeruginosa infections are not effective (3) as its outer membrane offers low permeability, 65 which limits the penetration of antibiotics into the bacterial cells thereby increase antibiotic 66 resistance (4, 5). Given the severity of P. aeruginosa infections and the limited antimicrobial 67 arsenal with which to treat them, finding new alternative antimicrobials with unique 68 mechanisms of action is urgently required (6). 69 Antimicr...

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Section: Discussionmentioning

confidence: 74%

Section: Methodsmentioning

confidence: 99%

Comparative mode of action of antimicrobial peptide melimine and its derivative Mel4 against Pseudomonas aeruginosa

Yasir

Dutta

Willcox

2018

Preprint

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“…The immunomodulating properties of RTD-1 were recently extended to a rat model of rheumatoid arthritis, wherein peptide treatment arrested and promoted the resolution of both evolving and severe joint disease (55). While RTD-1 is the first AMP shown to be effective in autoimmune arthritis, several AMPs, including ␣-defensins, ␤-defensins, and cathelicidin LL37, have been shown to have combined antimicrobial/immunomodulatory properties (27,28,(56)(57)(58)(59).…”

Section: Efficacy Of -Defensins Against Mdr Candida Speciesmentioning

confidence: 99%

Fungicidal Potency and Mechanisms of θ-Defensins against Multidrug-Resistant Candida Species

Basso

Garcia

Tran

et al. 2018

Antimicrob Agents Chemother

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Systemic candidiasis is a growing health care concern that is becoming even more challenging due to the growing frequency of infections caused by multidrug-resistant (MDR) species. Thus, there is an urgent need for new therapeutic approaches to candidiasis, including strategies bioinspired by insights into natural host defense against fungal pathogens. The antifungal properties of θ-defensins, macrocyclic peptides expressed in tissues of Old World monkeys, were investigated against a panel of drug-sensitive and drug-resistant clinical isolates of and non- species. Rhesus θ-defensin 1 (RTD-1), the prototype θ-defensin, was rapidly and potently fungicidal against drug-sensitive and MDR strains. Fungal killing occurred by cell permeabilization that was temporally correlated with ATP release and intracellular accumulation of reactive oxygen species (ROS). Killing by RTD-1 was compared with that by histatin 5 (Hst 5), an extensively characterized anticandidal peptide expressed in human saliva. RTD-1 killed much more rapidly and at a >200-fold lower concentration than that of Hst 5. Unlike Hst 5, the anticandidal activity of RTD-1 was independent of mitochondrial ATP production. Moreover, RTD-1 was completely resistant to proteases for 2 h under conditions that rapidly and completely degraded Hst 5. MICs and minimum fungicidal concentrations (MFCs) of 14 natural θ-defensins isoforms against drug-resistant isolates identified peptides that are more active than amphotericin B and/or caspofungin against fluconazole-resistant organisms, including MDR These results point to the potential of macrocyclic θ-defensins as structural templates for the design of antifungal therapeutics.

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“…PR‐39, a proline‐rich AMP of the cathelicidin family, has a broad antibacterial spectrum (Veldhuizen et al . ).…”

Section: Introductionmentioning

confidence: 97%

Comparison of gene expression of Toll-like receptors and antimicrobial peptides in immune organs and tissues between Yorkshire and Tibetan pigs

et al. 2015

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Toll-like receptors (TLRs), an important family of pattern-recognition receptors, and antimicrobial peptides (AMPs) contribute to the first line of innate protection of mammals against microbes. To compare characteristics of innate immunity between Tibetan and Yorkshire pigs, we investigated the mRNA abundance of TLR genes (TLR1-TLR9) and two AMP-encoding genes (PBD-1 and PR-39) in thymus, spleen, blood, palatine tonsils, and mesenteric and pulmonary hilar lymph nodes of the two breeds at ages of 6, 12 and 24 weeks using quantitative real-time PCR. Results showed that all mRNAs were detected in all tissues. Transcript levels of the major TLR genes of Tibetan pigs were significantly higher than those of Yorkshires in most tissues of the immune system, with a higher abundance of porcine (PBD-1) (beta-defensin-1) and PR-39 mRNA in lymphoid organs and tissues, especially blood, palatine tonsils, and mesenteric and pulmonary hilar lymph nodes. Our data suggest that Tibetan pigs have stronger innate immunity for triggering local and/or systemic immune responses to eliminate infections with pathogenic microorganisms.

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Antimicrobial and Immunomodulatory Activities of PR-39 Derived Peptides

Cited by 103 publications

References 39 publications

Comparative mode of action of antimicrobial peptide melimine and its derivative Mel4 against Pseudomonas aeruginosa

Comparative mode of action of antimicrobial peptide melimine and its derivative Mel4 against Pseudomonas aeruginosa

Fungicidal Potency and Mechanisms of θ-Defensins against Multidrug-Resistant Candida Species

Comparison of gene expression of Toll-like receptors and antimicrobial peptides in immune organs and tissues between Yorkshire and Tibetan pigs

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