Cargando…
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...
Autores principales: | , , , , |
---|---|
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 |
_version_ | 1782292753306091520 |
---|---|
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. |
format | Online Article Text |
id | pubmed-3841204 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT berishaarton highresolutionmassspectrometrydrivendiscoveryofpeptidicdangersignalsininsectimmunity AT mukherjeekrishnendu highresolutionmassspectrometrydrivendiscoveryofpeptidicdangersignalsininsectimmunity AT vilcinskasandreas highresolutionmassspectrometrydrivendiscoveryofpeptidicdangersignalsininsectimmunity AT spenglerbernhard highresolutionmassspectrometrydrivendiscoveryofpeptidicdangersignalsininsectimmunity AT romppandreas highresolutionmassspectrometrydrivendiscoveryofpeptidicdangersignalsininsectimmunity |