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High-Resolution Mass Spectrometry Driven Discovery of Peptidic Danger Signals in Insect Immunity

The ‘danger model’ is an alternative concept for immune response postulating that the immune system reacts to entities that do damage (danger associated molecular patterns, DAMP) and not only to entities that are foreign (pathogen-associated molecular patterns, PAMP) as proposed by classical immunol...

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Autores principales: Berisha, Arton, Mukherjee, Krishnendu, Vilcinskas, Andreas, Spengler, Bernhard, Römpp, Andreas
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3841204/
https://www.ncbi.nlm.nih.gov/pubmed/24303012
http://dx.doi.org/10.1371/journal.pone.0080406
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author Berisha, Arton
Mukherjee, Krishnendu
Vilcinskas, Andreas
Spengler, Bernhard
Römpp, Andreas
author_facet Berisha, Arton
Mukherjee, Krishnendu
Vilcinskas, Andreas
Spengler, Bernhard
Römpp, Andreas
author_sort Berisha, Arton
collection PubMed
description The ‘danger model’ is an alternative concept for immune response postulating that the immune system reacts to entities that do damage (danger associated molecular patterns, DAMP) and not only to entities that are foreign (pathogen-associated molecular patterns, PAMP) as proposed by classical immunology concepts. In this study we used Galleria mellonella to validate the danger model in insects. Hemolymph of G. mellonella was digested with thermolysin (as a representative for virulence-associated metalloproteinases produced by humanpathogens) followed by chromatographic fractionation. Immune-stimulatory activity was tested by measuring lysozyme activity with the lytic zone assays against Micrococcus luteus cell wall components. Peptides were analyzed by nano-scale liquid chromatography coupled to high-resolution Fourier transform mass spectrometers. Addressing the lack of a genome sequence we complemented the rudimentary NCBI protein database with a recently established transcriptome and de novo sequencing methods for peptide identification. This approach led to identification of 127 peptides, 9 of which were identified in bioactive fractions. Detailed MS/MS experiments in comparison with synthetic analogues confirmed the amino acid sequence of all 9 peptides. To test the potential of these putative danger signals to induce immune responses we injected the synthetic analogues into G. mellonella and monitored the anti-bacterial activity against living Micrococcus luteus. Six out of 9 peptides identified in the bioactive fractions exhibited immune-stimulatory activity when injected. Hence, we provide evidence that small peptides resulting from thermolysin-mediated digestion of hemolymph proteins function as endogenous danger signals which can set the immune system into alarm. Consequently, our study indicates that the danger model also plays a role in insect immunity.
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spelling pubmed-38412042013-12-03 High-Resolution Mass Spectrometry Driven Discovery of Peptidic Danger Signals in Insect Immunity Berisha, Arton Mukherjee, Krishnendu Vilcinskas, Andreas Spengler, Bernhard Römpp, Andreas PLoS One Research Article The ‘danger model’ is an alternative concept for immune response postulating that the immune system reacts to entities that do damage (danger associated molecular patterns, DAMP) and not only to entities that are foreign (pathogen-associated molecular patterns, PAMP) as proposed by classical immunology concepts. In this study we used Galleria mellonella to validate the danger model in insects. Hemolymph of G. mellonella was digested with thermolysin (as a representative for virulence-associated metalloproteinases produced by humanpathogens) followed by chromatographic fractionation. Immune-stimulatory activity was tested by measuring lysozyme activity with the lytic zone assays against Micrococcus luteus cell wall components. Peptides were analyzed by nano-scale liquid chromatography coupled to high-resolution Fourier transform mass spectrometers. Addressing the lack of a genome sequence we complemented the rudimentary NCBI protein database with a recently established transcriptome and de novo sequencing methods for peptide identification. This approach led to identification of 127 peptides, 9 of which were identified in bioactive fractions. Detailed MS/MS experiments in comparison with synthetic analogues confirmed the amino acid sequence of all 9 peptides. To test the potential of these putative danger signals to induce immune responses we injected the synthetic analogues into G. mellonella and monitored the anti-bacterial activity against living Micrococcus luteus. Six out of 9 peptides identified in the bioactive fractions exhibited immune-stimulatory activity when injected. Hence, we provide evidence that small peptides resulting from thermolysin-mediated digestion of hemolymph proteins function as endogenous danger signals which can set the immune system into alarm. Consequently, our study indicates that the danger model also plays a role in insect immunity. Public Library of Science 2013-11-26 /pmc/articles/PMC3841204/ /pubmed/24303012 http://dx.doi.org/10.1371/journal.pone.0080406 Text en © 2013 Berisha et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Berisha, Arton
Mukherjee, Krishnendu
Vilcinskas, Andreas
Spengler, Bernhard
Römpp, Andreas
High-Resolution Mass Spectrometry Driven Discovery of Peptidic Danger Signals in Insect Immunity
title High-Resolution Mass Spectrometry Driven Discovery of Peptidic Danger Signals in Insect Immunity
title_full High-Resolution Mass Spectrometry Driven Discovery of Peptidic Danger Signals in Insect Immunity
title_fullStr High-Resolution Mass Spectrometry Driven Discovery of Peptidic Danger Signals in Insect Immunity
title_full_unstemmed High-Resolution Mass Spectrometry Driven Discovery of Peptidic Danger Signals in Insect Immunity
title_short High-Resolution Mass Spectrometry Driven Discovery of Peptidic Danger Signals in Insect Immunity
title_sort high-resolution mass spectrometry driven discovery of peptidic danger signals in insect immunity
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3841204/
https://www.ncbi.nlm.nih.gov/pubmed/24303012
http://dx.doi.org/10.1371/journal.pone.0080406
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