Towards High-throughput Immunomics for Infectious Diseases: Use of Next-generation Peptide Microarrays for Rapid Discovery and Mapping of Antigenic Determinants.
Identifieur interne : 001299 ( PubMed/Checkpoint ); précédent : 001298; suivant : 001300Towards High-throughput Immunomics for Infectious Diseases: Use of Next-generation Peptide Microarrays for Rapid Discovery and Mapping of Antigenic Determinants.
Auteurs : Santiago J. Carmona [Argentine] ; Morten Nielsen [Danemark] ; Claus Schafer-Nielsen [Danemark] ; Juan Mucci [Argentine] ; Jaime Altcheh [Argentine] ; Virginia Balouz [Argentine] ; Valeria Tekiel [Argentine] ; Alberto C. Frasch [Argentine] ; Oscar Campetella [Argentine] ; Carlos A. Buscaglia [Argentine] ; Fernán Agüero [Argentine]Source :
- Molecular & cellular proteomics : MCP [ 1535-9484 ] ; 2015.
Descripteurs français
- KwdFr :
- Analyse par réseau de protéines (), Antigènes de protozoaire (immunologie), Bases de données de protéines, Cartographie épitopique (), Déterminants antigéniques des lymphocytes B (immunologie), Humains, Lymphocytes B (immunologie), Maladie de Chagas (immunologie), Peptides (métabolisme), Protéomique (), Reproductibilité des résultats, Test ELISA, Tests de criblage à haut débit ().
- MESH :
- immunologie : Antigènes de protozoaire, Déterminants antigéniques des lymphocytes B, Lymphocytes B, Maladie de Chagas.
- métabolisme : Peptides.
- Analyse par réseau de protéines, Bases de données de protéines, Cartographie épitopique, Humains, Protéomique, Reproductibilité des résultats, Test ELISA, Tests de criblage à haut débit.
English descriptors
- KwdEn :
- Antigens, Protozoan (immunology), B-Lymphocytes (immunology), Chagas Disease (immunology), Databases, Protein, Enzyme-Linked Immunosorbent Assay, Epitope Mapping (methods), Epitopes, B-Lymphocyte (immunology), High-Throughput Screening Assays (methods), Humans, Peptides (metabolism), Protein Array Analysis (methods), Proteomics (methods), Reproducibility of Results.
- MESH :
- chemical , immunology : Antigens, Protozoan, Epitopes, B-Lymphocyte.
- immunology : B-Lymphocytes, Chagas Disease.
- chemical , metabolism : Peptides.
- methods : Epitope Mapping, High-Throughput Screening Assays, Protein Array Analysis, Proteomics.
- Databases, Protein, Enzyme-Linked Immunosorbent Assay, Humans, Reproducibility of Results.
Abstract
Complete characterization of antibody specificities associated to natural infections is expected to provide a rich source of serologic biomarkers with potential applications in molecular diagnosis, follow-up of chemotherapeutic treatments, and prioritization of targets for vaccine development. Here, we developed a highly-multiplexed platform based on next-generation high-density peptide microarrays to map these specificities in Chagas Disease, an exemplar of a human infectious disease caused by the protozoan Trypanosoma cruzi. We designed a high-density peptide microarray containing more than 175,000 overlapping 15 mer peptides derived from T. cruzi proteins. Peptides were synthesized in situ on microarray slides, spanning the complete length of 457 parasite proteins with fully overlapped 15 mers (1 residue shift). Screening of these slides with antibodies purified from infected patients and healthy donors demonstrated both a high technical reproducibility as well as epitope mapping consistency when compared with earlier low-throughput technologies. Using a conservative signal threshold to classify positive (reactive) peptides we identified 2,031 disease-specific peptides and 97 novel parasite antigens, effectively doubling the number of known antigens and providing a 10-fold increase in the number of fine mapped antigenic determinants for this disease. Finally, further analysis of the chip data showed that optimizing the amount of sequence overlap of displayed peptides can increase the protein space covered in a single chip by at least ∼ threefold without sacrificing sensitivity. In conclusion, we show the power of high-density peptide chips for the discovery of pathogen-specific linear B-cell epitopes from clinical samples, thus setting the stage for high-throughput biomarker discovery screenings and proteome-wide studies of immune responses against pathogens.
DOI: 10.1074/mcp.M114.045906
PubMed: 25922409
Affiliations:
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Carmona</name><affiliation wicri:level="1"><nlm:affiliation>From the ‡Instituto de Investigaciones Biotecnológicas - Instituto Tecnológico de Chascomús, Universidad de San Martín - CONICET, Sede San Martín, B 1650 HMP, San Martín, Buenos Aires, Argentina;</nlm:affiliation><country xml:lang="fr">Argentine</country><wicri:regionArea>From the ‡Instituto de Investigaciones Biotecnológicas - Instituto Tecnológico de Chascomús, Universidad de San Martín - CONICET, Sede San Martín, B 1650 HMP, San Martín, Buenos Aires</wicri:regionArea><wicri:noRegion>Buenos Aires</wicri:noRegion></affiliation></author><author><name sortKey="Nielsen, Morten" sort="Nielsen, Morten" uniqKey="Nielsen M" first="Morten" last="Nielsen">Morten Nielsen</name><affiliation wicri:level="1"><nlm:affiliation>From the ‡Instituto de Investigaciones Biotecnológicas - Instituto Tecnológico de Chascomús, Universidad de San Martín - CONICET, Sede San Martín, B 1650 HMP, San Martín, Buenos Aires, Argentina; §Center for Biological Sequence Analysis, Department of Systems Biology, Technical University of Denmark, 2800 Lyngby, Denmark;</nlm:affiliation><country xml:lang="fr">Danemark</country><wicri:regionArea>From the ‡Instituto de Investigaciones Biotecnológicas - Instituto Tecnológico de Chascomús, Universidad de San Martín - CONICET, Sede San Martín, B 1650 HMP, San Martín, Buenos Aires, Argentina; §Center for Biological Sequence Analysis, Department of Systems Biology, Technical University of Denmark, 2800 Lyngby</wicri:regionArea><wicri:noRegion>2800 Lyngby</wicri:noRegion></affiliation></author><author><name sortKey="Schafer Nielsen, Claus" sort="Schafer Nielsen, Claus" uniqKey="Schafer Nielsen C" first="Claus" last="Schafer-Nielsen">Claus Schafer-Nielsen</name><affiliation wicri:level="1"><nlm:affiliation>¶Schafer-N ApS, 2100 Copenhagen, Denmark;</nlm:affiliation><country xml:lang="fr">Danemark</country><wicri:regionArea>¶Schafer-N ApS, 2100 Copenhagen</wicri:regionArea><placeName><settlement type="city">Copenhague</settlement><region type="région" nuts="2">Hovedstaden</region></placeName></affiliation></author><author><name sortKey="Mucci, Juan" sort="Mucci, Juan" uniqKey="Mucci J" first="Juan" last="Mucci">Juan Mucci</name><affiliation wicri:level="1"><nlm:affiliation>From the ‡Instituto de Investigaciones Biotecnológicas - Instituto Tecnológico de Chascomús, Universidad de San Martín - CONICET, Sede San Martín, B 1650 HMP, San Martín, Buenos Aires, Argentina;</nlm:affiliation><country xml:lang="fr">Argentine</country><wicri:regionArea>From the ‡Instituto de Investigaciones Biotecnológicas - Instituto Tecnológico de Chascomús, Universidad de San Martín - CONICET, Sede San Martín, B 1650 HMP, San Martín, Buenos Aires</wicri:regionArea><wicri:noRegion>Buenos Aires</wicri:noRegion></affiliation></author><author><name sortKey="Altcheh, Jaime" sort="Altcheh, Jaime" uniqKey="Altcheh J" first="Jaime" last="Altcheh">Jaime Altcheh</name><affiliation wicri:level="1"><nlm:affiliation>‖Servicio de Parasitología y Chagas, Hospital de Niños Ricardo Gutiérrez, Ciudad de Buenos Aires, Argentina.</nlm:affiliation><country xml:lang="fr">Argentine</country><wicri:regionArea>‖Servicio de Parasitología y Chagas, Hospital de Niños Ricardo Gutiérrez, Ciudad de Buenos Aires</wicri:regionArea><wicri:noRegion>Ciudad de Buenos Aires</wicri:noRegion></affiliation></author><author><name sortKey="Balouz, Virginia" sort="Balouz, Virginia" uniqKey="Balouz V" first="Virginia" last="Balouz">Virginia Balouz</name><affiliation wicri:level="1"><nlm:affiliation>From the ‡Instituto de Investigaciones Biotecnológicas - Instituto Tecnológico de Chascomús, Universidad de San Martín - CONICET, Sede San Martín, B 1650 HMP, San Martín, Buenos Aires, Argentina;</nlm:affiliation><country xml:lang="fr">Argentine</country><wicri:regionArea>From the ‡Instituto de Investigaciones Biotecnológicas - Instituto Tecnológico de Chascomús, Universidad de San Martín - CONICET, Sede San Martín, B 1650 HMP, San Martín, Buenos Aires</wicri:regionArea><wicri:noRegion>Buenos Aires</wicri:noRegion></affiliation></author><author><name sortKey="Tekiel, Valeria" sort="Tekiel, Valeria" uniqKey="Tekiel V" first="Valeria" last="Tekiel">Valeria Tekiel</name><affiliation wicri:level="1"><nlm:affiliation>From the ‡Instituto de Investigaciones Biotecnológicas - Instituto Tecnológico de Chascomús, Universidad de San Martín - CONICET, Sede San Martín, B 1650 HMP, San Martín, Buenos Aires, Argentina;</nlm:affiliation><country xml:lang="fr">Argentine</country><wicri:regionArea>From the ‡Instituto de Investigaciones Biotecnológicas - Instituto Tecnológico de Chascomús, Universidad de San Martín - CONICET, Sede San Martín, B 1650 HMP, San Martín, Buenos Aires</wicri:regionArea><wicri:noRegion>Buenos Aires</wicri:noRegion></affiliation></author><author><name sortKey="Frasch, Alberto C" sort="Frasch, Alberto C" uniqKey="Frasch A" first="Alberto C" last="Frasch">Alberto C. Frasch</name><affiliation wicri:level="1"><nlm:affiliation>From the ‡Instituto de Investigaciones Biotecnológicas - Instituto Tecnológico de Chascomús, Universidad de San Martín - CONICET, Sede San Martín, B 1650 HMP, San Martín, Buenos Aires, Argentina;</nlm:affiliation><country xml:lang="fr">Argentine</country><wicri:regionArea>From the ‡Instituto de Investigaciones Biotecnológicas - Instituto Tecnológico de Chascomús, Universidad de San Martín - CONICET, Sede San Martín, B 1650 HMP, San Martín, Buenos Aires</wicri:regionArea><wicri:noRegion>Buenos Aires</wicri:noRegion></affiliation></author><author><name sortKey="Campetella, Oscar" sort="Campetella, Oscar" uniqKey="Campetella O" first="Oscar" last="Campetella">Oscar Campetella</name><affiliation wicri:level="1"><nlm:affiliation>From the ‡Instituto de Investigaciones Biotecnológicas - Instituto Tecnológico de Chascomús, Universidad de San Martín - CONICET, Sede San Martín, B 1650 HMP, San Martín, Buenos Aires, Argentina;</nlm:affiliation><country xml:lang="fr">Argentine</country><wicri:regionArea>From the ‡Instituto de Investigaciones Biotecnológicas - Instituto Tecnológico de Chascomús, Universidad de San Martín - CONICET, Sede San Martín, B 1650 HMP, San Martín, Buenos Aires</wicri:regionArea><wicri:noRegion>Buenos Aires</wicri:noRegion></affiliation></author><author><name sortKey="Buscaglia, Carlos A" sort="Buscaglia, Carlos A" uniqKey="Buscaglia C" first="Carlos A" last="Buscaglia">Carlos A. Buscaglia</name><affiliation wicri:level="1"><nlm:affiliation>From the ‡Instituto de Investigaciones Biotecnológicas - Instituto Tecnológico de Chascomús, Universidad de San Martín - CONICET, Sede San Martín, B 1650 HMP, San Martín, Buenos Aires, Argentina;</nlm:affiliation><country xml:lang="fr">Argentine</country><wicri:regionArea>From the ‡Instituto de Investigaciones Biotecnológicas - Instituto Tecnológico de Chascomús, Universidad de San Martín - CONICET, Sede San Martín, B 1650 HMP, San Martín, Buenos Aires</wicri:regionArea><wicri:noRegion>Buenos Aires</wicri:noRegion></affiliation></author><author><name sortKey="Aguero, Fernan" sort="Aguero, Fernan" uniqKey="Aguero F" first="Fernán" last="Agüero">Fernán Agüero</name><affiliation wicri:level="1"><nlm:affiliation>From the ‡Instituto de Investigaciones Biotecnológicas - Instituto Tecnológico de Chascomús, Universidad de San Martín - CONICET, Sede San Martín, B 1650 HMP, San Martín, Buenos Aires, Argentina; fernan@unsam.edu.ar.</nlm:affiliation><country wicri:rule="url">Argentine</country><wicri:regionArea>From the ‡Instituto de Investigaciones Biotecnológicas - Instituto Tecnológico de Chascomús, Universidad de San Martín - CONICET, Sede San Martín, B 1650 HMP, San Martín, Buenos Aires</wicri:regionArea><wicri:noRegion>Buenos Aires</wicri:noRegion></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2015">2015</date><idno type="RBID">pubmed:25922409</idno><idno type="pmid">25922409</idno><idno type="doi">10.1074/mcp.M114.045906</idno><idno type="wicri:Area/PubMed/Corpus">001633</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">001633</idno><idno type="wicri:Area/PubMed/Curation">001633</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Curation">001633</idno><idno type="wicri:Area/PubMed/Checkpoint">001299</idno><idno type="wicri:explorRef" wicri:stream="Checkpoint" wicri:step="PubMed">001299</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Towards High-throughput Immunomics for Infectious Diseases: Use of Next-generation Peptide Microarrays for Rapid Discovery and Mapping of Antigenic Determinants.</title><author><name sortKey="Carmona, Santiago J" sort="Carmona, Santiago J" uniqKey="Carmona S" first="Santiago J" last="Carmona">Santiago J. Carmona</name><affiliation wicri:level="1"><nlm:affiliation>From the ‡Instituto de Investigaciones Biotecnológicas - Instituto Tecnológico de Chascomús, Universidad de San Martín - CONICET, Sede San Martín, B 1650 HMP, San Martín, Buenos Aires, Argentina;</nlm:affiliation><country xml:lang="fr">Argentine</country><wicri:regionArea>From the ‡Instituto de Investigaciones Biotecnológicas - Instituto Tecnológico de Chascomús, Universidad de San Martín - CONICET, Sede San Martín, B 1650 HMP, San Martín, Buenos Aires</wicri:regionArea><wicri:noRegion>Buenos Aires</wicri:noRegion></affiliation></author><author><name sortKey="Nielsen, Morten" sort="Nielsen, Morten" uniqKey="Nielsen M" first="Morten" last="Nielsen">Morten Nielsen</name><affiliation wicri:level="1"><nlm:affiliation>From the ‡Instituto de Investigaciones Biotecnológicas - Instituto Tecnológico de Chascomús, Universidad de San Martín - CONICET, Sede San Martín, B 1650 HMP, San Martín, Buenos Aires, Argentina; §Center for Biological Sequence Analysis, Department of Systems Biology, Technical University of Denmark, 2800 Lyngby, Denmark;</nlm:affiliation><country xml:lang="fr">Danemark</country><wicri:regionArea>From the ‡Instituto de Investigaciones Biotecnológicas - Instituto Tecnológico de Chascomús, Universidad de San Martín - CONICET, Sede San Martín, B 1650 HMP, San Martín, Buenos Aires, Argentina; §Center for Biological Sequence Analysis, Department of Systems Biology, Technical University of Denmark, 2800 Lyngby</wicri:regionArea><wicri:noRegion>2800 Lyngby</wicri:noRegion></affiliation></author><author><name sortKey="Schafer Nielsen, Claus" sort="Schafer Nielsen, Claus" uniqKey="Schafer Nielsen C" first="Claus" last="Schafer-Nielsen">Claus Schafer-Nielsen</name><affiliation wicri:level="1"><nlm:affiliation>¶Schafer-N ApS, 2100 Copenhagen, Denmark;</nlm:affiliation><country xml:lang="fr">Danemark</country><wicri:regionArea>¶Schafer-N ApS, 2100 Copenhagen</wicri:regionArea><placeName><settlement type="city">Copenhague</settlement><region type="région" nuts="2">Hovedstaden</region></placeName></affiliation></author><author><name sortKey="Mucci, Juan" sort="Mucci, Juan" uniqKey="Mucci J" first="Juan" last="Mucci">Juan Mucci</name><affiliation wicri:level="1"><nlm:affiliation>From the ‡Instituto de Investigaciones Biotecnológicas - Instituto Tecnológico de Chascomús, Universidad de San Martín - CONICET, Sede San Martín, B 1650 HMP, San Martín, Buenos Aires, Argentina;</nlm:affiliation><country xml:lang="fr">Argentine</country><wicri:regionArea>From the ‡Instituto de Investigaciones Biotecnológicas - Instituto Tecnológico de Chascomús, Universidad de San Martín - CONICET, Sede San Martín, B 1650 HMP, San Martín, Buenos Aires</wicri:regionArea><wicri:noRegion>Buenos Aires</wicri:noRegion></affiliation></author><author><name sortKey="Altcheh, Jaime" sort="Altcheh, Jaime" uniqKey="Altcheh J" first="Jaime" last="Altcheh">Jaime Altcheh</name><affiliation wicri:level="1"><nlm:affiliation>‖Servicio de Parasitología y Chagas, Hospital de Niños Ricardo Gutiérrez, Ciudad de Buenos Aires, Argentina.</nlm:affiliation><country xml:lang="fr">Argentine</country><wicri:regionArea>‖Servicio de Parasitología y Chagas, Hospital de Niños Ricardo Gutiérrez, Ciudad de Buenos Aires</wicri:regionArea><wicri:noRegion>Ciudad de Buenos Aires</wicri:noRegion></affiliation></author><author><name sortKey="Balouz, Virginia" sort="Balouz, Virginia" uniqKey="Balouz V" first="Virginia" last="Balouz">Virginia Balouz</name><affiliation wicri:level="1"><nlm:affiliation>From the ‡Instituto de Investigaciones Biotecnológicas - Instituto Tecnológico de Chascomús, Universidad de San Martín - CONICET, Sede San Martín, B 1650 HMP, San Martín, Buenos Aires, Argentina;</nlm:affiliation><country xml:lang="fr">Argentine</country><wicri:regionArea>From the ‡Instituto de Investigaciones Biotecnológicas - Instituto Tecnológico de Chascomús, Universidad de San Martín - CONICET, Sede San Martín, B 1650 HMP, San Martín, Buenos Aires</wicri:regionArea><wicri:noRegion>Buenos Aires</wicri:noRegion></affiliation></author><author><name sortKey="Tekiel, Valeria" sort="Tekiel, Valeria" uniqKey="Tekiel V" first="Valeria" last="Tekiel">Valeria Tekiel</name><affiliation wicri:level="1"><nlm:affiliation>From the ‡Instituto de Investigaciones Biotecnológicas - Instituto Tecnológico de Chascomús, Universidad de San Martín - CONICET, Sede San Martín, B 1650 HMP, San Martín, Buenos Aires, Argentina;</nlm:affiliation><country xml:lang="fr">Argentine</country><wicri:regionArea>From the ‡Instituto de Investigaciones Biotecnológicas - Instituto Tecnológico de Chascomús, Universidad de San Martín - CONICET, Sede San Martín, B 1650 HMP, San Martín, Buenos Aires</wicri:regionArea><wicri:noRegion>Buenos Aires</wicri:noRegion></affiliation></author><author><name sortKey="Frasch, Alberto C" sort="Frasch, Alberto C" uniqKey="Frasch A" first="Alberto C" last="Frasch">Alberto C. Frasch</name><affiliation wicri:level="1"><nlm:affiliation>From the ‡Instituto de Investigaciones Biotecnológicas - Instituto Tecnológico de Chascomús, Universidad de San Martín - CONICET, Sede San Martín, B 1650 HMP, San Martín, Buenos Aires, Argentina;</nlm:affiliation><country xml:lang="fr">Argentine</country><wicri:regionArea>From the ‡Instituto de Investigaciones Biotecnológicas - Instituto Tecnológico de Chascomús, Universidad de San Martín - CONICET, Sede San Martín, B 1650 HMP, San Martín, Buenos Aires</wicri:regionArea><wicri:noRegion>Buenos Aires</wicri:noRegion></affiliation></author><author><name sortKey="Campetella, Oscar" sort="Campetella, Oscar" uniqKey="Campetella O" first="Oscar" last="Campetella">Oscar Campetella</name><affiliation wicri:level="1"><nlm:affiliation>From the ‡Instituto de Investigaciones Biotecnológicas - Instituto Tecnológico de Chascomús, Universidad de San Martín - CONICET, Sede San Martín, B 1650 HMP, San Martín, Buenos Aires, Argentina;</nlm:affiliation><country xml:lang="fr">Argentine</country><wicri:regionArea>From the ‡Instituto de Investigaciones Biotecnológicas - Instituto Tecnológico de Chascomús, Universidad de San Martín - CONICET, Sede San Martín, B 1650 HMP, San Martín, Buenos Aires</wicri:regionArea><wicri:noRegion>Buenos Aires</wicri:noRegion></affiliation></author><author><name sortKey="Buscaglia, Carlos A" sort="Buscaglia, Carlos A" uniqKey="Buscaglia C" first="Carlos A" last="Buscaglia">Carlos A. Buscaglia</name><affiliation wicri:level="1"><nlm:affiliation>From the ‡Instituto de Investigaciones Biotecnológicas - Instituto Tecnológico de Chascomús, Universidad de San Martín - CONICET, Sede San Martín, B 1650 HMP, San Martín, Buenos Aires, Argentina;</nlm:affiliation><country xml:lang="fr">Argentine</country><wicri:regionArea>From the ‡Instituto de Investigaciones Biotecnológicas - Instituto Tecnológico de Chascomús, Universidad de San Martín - CONICET, Sede San Martín, B 1650 HMP, San Martín, Buenos Aires</wicri:regionArea><wicri:noRegion>Buenos Aires</wicri:noRegion></affiliation></author><author><name sortKey="Aguero, Fernan" sort="Aguero, Fernan" uniqKey="Aguero F" first="Fernán" last="Agüero">Fernán Agüero</name><affiliation wicri:level="1"><nlm:affiliation>From the ‡Instituto de Investigaciones Biotecnológicas - Instituto Tecnológico de Chascomús, Universidad de San Martín - CONICET, Sede San Martín, B 1650 HMP, San Martín, Buenos Aires, Argentina; fernan@unsam.edu.ar.</nlm:affiliation><country wicri:rule="url">Argentine</country><wicri:regionArea>From the ‡Instituto de Investigaciones Biotecnológicas - Instituto Tecnológico de Chascomús, Universidad de San Martín - CONICET, Sede San Martín, B 1650 HMP, San Martín, Buenos Aires</wicri:regionArea><wicri:noRegion>Buenos Aires</wicri:noRegion></affiliation></author></analytic><series><title level="j">Molecular & cellular proteomics : MCP</title><idno type="eISSN">1535-9484</idno><imprint><date when="2015" type="published">2015</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Antigens, Protozoan (immunology)</term><term>B-Lymphocytes (immunology)</term><term>Chagas Disease (immunology)</term><term>Databases, Protein</term><term>Enzyme-Linked Immunosorbent Assay</term><term>Epitope Mapping (methods)</term><term>Epitopes, B-Lymphocyte (immunology)</term><term>High-Throughput Screening Assays (methods)</term><term>Humans</term><term>Peptides (metabolism)</term><term>Protein Array Analysis (methods)</term><term>Proteomics (methods)</term><term>Reproducibility of Results</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Analyse par réseau de protéines ()</term><term>Antigènes de protozoaire (immunologie)</term><term>Bases de données de protéines</term><term>Cartographie épitopique ()</term><term>Déterminants antigéniques des lymphocytes B (immunologie)</term><term>Humains</term><term>Lymphocytes B (immunologie)</term><term>Maladie de Chagas (immunologie)</term><term>Peptides (métabolisme)</term><term>Protéomique ()</term><term>Reproductibilité des résultats</term><term>Test ELISA</term><term>Tests de criblage à haut débit ()</term></keywords><keywords scheme="MESH" type="chemical" qualifier="immunology" xml:lang="en"><term>Antigens, Protozoan</term><term>Epitopes, B-Lymphocyte</term></keywords><keywords scheme="MESH" qualifier="immunologie" xml:lang="fr"><term>Antigènes de protozoaire</term><term>Déterminants antigéniques des lymphocytes B</term><term>Lymphocytes B</term><term>Maladie de Chagas</term></keywords><keywords scheme="MESH" qualifier="immunology" xml:lang="en"><term>B-Lymphocytes</term><term>Chagas Disease</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Peptides</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Epitope Mapping</term><term>High-Throughput Screening Assays</term><term>Protein Array Analysis</term><term>Proteomics</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Peptides</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Databases, Protein</term><term>Enzyme-Linked Immunosorbent Assay</term><term>Humans</term><term>Reproducibility of Results</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Analyse par réseau de protéines</term><term>Bases de données de protéines</term><term>Cartographie épitopique</term><term>Humains</term><term>Protéomique</term><term>Reproductibilité des résultats</term><term>Test ELISA</term><term>Tests de criblage à haut débit</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Complete characterization of antibody specificities associated to natural infections is expected to provide a rich source of serologic biomarkers with potential applications in molecular diagnosis, follow-up of chemotherapeutic treatments, and prioritization of targets for vaccine development. Here, we developed a highly-multiplexed platform based on next-generation high-density peptide microarrays to map these specificities in Chagas Disease, an exemplar of a human infectious disease caused by the protozoan Trypanosoma cruzi. We designed a high-density peptide microarray containing more than 175,000 overlapping 15 mer peptides derived from T. cruzi proteins. Peptides were synthesized in situ on microarray slides, spanning the complete length of 457 parasite proteins with fully overlapped 15 mers (1 residue shift). Screening of these slides with antibodies purified from infected patients and healthy donors demonstrated both a high technical reproducibility as well as epitope mapping consistency when compared with earlier low-throughput technologies. Using a conservative signal threshold to classify positive (reactive) peptides we identified 2,031 disease-specific peptides and 97 novel parasite antigens, effectively doubling the number of known antigens and providing a 10-fold increase in the number of fine mapped antigenic determinants for this disease. Finally, further analysis of the chip data showed that optimizing the amount of sequence overlap of displayed peptides can increase the protein space covered in a single chip by at least ∼ threefold without sacrificing sensitivity. In conclusion, we show the power of high-density peptide chips for the discovery of pathogen-specific linear B-cell epitopes from clinical samples, thus setting the stage for high-throughput biomarker discovery screenings and proteome-wide studies of immune responses against pathogens.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">25922409</PMID><DateCompleted><Year>2016</Year><Month>04</Month><Day>05</Day></DateCompleted><DateRevised><Year>2018</Year><Month>11</Month><Day>13</Day></DateRevised><Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Electronic">1535-9484</ISSN><JournalIssue CitedMedium="Internet"><Volume>14</Volume><Issue>7</Issue><PubDate><Year>2015</Year><Month>Jul</Month></PubDate></JournalIssue><Title>Molecular & cellular proteomics : MCP</Title><ISOAbbreviation>Mol. Cell Proteomics</ISOAbbreviation></Journal><ArticleTitle>Towards High-throughput Immunomics for Infectious Diseases: Use of Next-generation Peptide Microarrays for Rapid Discovery and Mapping of Antigenic Determinants.</ArticleTitle><Pagination><MedlinePgn>1871-84</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1074/mcp.M114.045906</ELocationID><Abstract><AbstractText>Complete characterization of antibody specificities associated to natural infections is expected to provide a rich source of serologic biomarkers with potential applications in molecular diagnosis, follow-up of chemotherapeutic treatments, and prioritization of targets for vaccine development. Here, we developed a highly-multiplexed platform based on next-generation high-density peptide microarrays to map these specificities in Chagas Disease, an exemplar of a human infectious disease caused by the protozoan Trypanosoma cruzi. We designed a high-density peptide microarray containing more than 175,000 overlapping 15 mer peptides derived from T. cruzi proteins. Peptides were synthesized in situ on microarray slides, spanning the complete length of 457 parasite proteins with fully overlapped 15 mers (1 residue shift). Screening of these slides with antibodies purified from infected patients and healthy donors demonstrated both a high technical reproducibility as well as epitope mapping consistency when compared with earlier low-throughput technologies. Using a conservative signal threshold to classify positive (reactive) peptides we identified 2,031 disease-specific peptides and 97 novel parasite antigens, effectively doubling the number of known antigens and providing a 10-fold increase in the number of fine mapped antigenic determinants for this disease. Finally, further analysis of the chip data showed that optimizing the amount of sequence overlap of displayed peptides can increase the protein space covered in a single chip by at least ∼ threefold without sacrificing sensitivity. In conclusion, we show the power of high-density peptide chips for the discovery of pathogen-specific linear B-cell epitopes from clinical samples, thus setting the stage for high-throughput biomarker discovery screenings and proteome-wide studies of immune responses against pathogens.</AbstractText><CopyrightInformation>© 2015 by The American Society for Biochemistry and Molecular Biology, Inc.</CopyrightInformation></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Carmona</LastName><ForeName>Santiago J</ForeName><Initials>SJ</Initials><AffiliationInfo><Affiliation>From the ‡Instituto de Investigaciones Biotecnológicas - Instituto Tecnológico de Chascomús, Universidad de San Martín - CONICET, Sede San Martín, B 1650 HMP, San Martín, Buenos Aires, Argentina;</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Nielsen</LastName><ForeName>Morten</ForeName><Initials>M</Initials><AffiliationInfo><Affiliation>From the ‡Instituto de Investigaciones Biotecnológicas - Instituto Tecnológico de Chascomús, Universidad de San Martín - CONICET, Sede San Martín, B 1650 HMP, San Martín, Buenos Aires, Argentina; §Center for Biological Sequence Analysis, Department of Systems Biology, Technical University of Denmark, 2800 Lyngby, Denmark;</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Schafer-Nielsen</LastName><ForeName>Claus</ForeName><Initials>C</Initials><AffiliationInfo><Affiliation>¶Schafer-N ApS, 2100 Copenhagen, Denmark;</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Mucci</LastName><ForeName>Juan</ForeName><Initials>J</Initials><AffiliationInfo><Affiliation>From the ‡Instituto de Investigaciones Biotecnológicas - Instituto Tecnológico de Chascomús, Universidad de San Martín - CONICET, Sede San Martín, B 1650 HMP, San Martín, Buenos Aires, Argentina;</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Altcheh</LastName><ForeName>Jaime</ForeName><Initials>J</Initials><AffiliationInfo><Affiliation>‖Servicio de Parasitología y Chagas, Hospital de Niños Ricardo Gutiérrez, Ciudad de Buenos Aires, Argentina.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Balouz</LastName><ForeName>Virginia</ForeName><Initials>V</Initials><AffiliationInfo><Affiliation>From the ‡Instituto de Investigaciones Biotecnológicas - Instituto Tecnológico de Chascomús, Universidad de San Martín - CONICET, Sede San Martín, B 1650 HMP, San Martín, Buenos Aires, Argentina;</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Tekiel</LastName><ForeName>Valeria</ForeName><Initials>V</Initials><AffiliationInfo><Affiliation>From the ‡Instituto de Investigaciones Biotecnológicas - Instituto Tecnológico de Chascomús, Universidad de San Martín - CONICET, Sede San Martín, B 1650 HMP, San Martín, Buenos Aires, Argentina;</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Frasch</LastName><ForeName>Alberto C</ForeName><Initials>AC</Initials><AffiliationInfo><Affiliation>From the ‡Instituto de Investigaciones Biotecnológicas - Instituto Tecnológico de Chascomús, Universidad de San Martín - CONICET, Sede San Martín, B 1650 HMP, San Martín, Buenos Aires, Argentina;</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Campetella</LastName><ForeName>Oscar</ForeName><Initials>O</Initials><AffiliationInfo><Affiliation>From the ‡Instituto de Investigaciones Biotecnológicas - Instituto Tecnológico de Chascomús, Universidad de San Martín - CONICET, Sede San Martín, B 1650 HMP, San Martín, Buenos Aires, Argentina;</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Buscaglia</LastName><ForeName>Carlos A</ForeName><Initials>CA</Initials><AffiliationInfo><Affiliation>From the ‡Instituto de Investigaciones Biotecnológicas - Instituto Tecnológico de Chascomús, Universidad de San Martín - CONICET, Sede San Martín, B 1650 HMP, San Martín, Buenos Aires, Argentina;</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Agüero</LastName><ForeName>Fernán</ForeName><Initials>F</Initials><AffiliationInfo><Affiliation>From the ‡Instituto de Investigaciones Biotecnológicas - Instituto Tecnológico de Chascomús, Universidad de San Martín - CONICET, Sede San Martín, B 1650 HMP, San Martín, Buenos Aires, Argentina; fernan@unsam.edu.ar.</Affiliation></AffiliationInfo></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType><PublicationType UI="D013485">Research Support, Non-U.S. Gov't</PublicationType></PublicationTypeList><ArticleDate DateType="Electronic"><Year>2015</Year><Month>04</Month><Day>28</Day></ArticleDate></Article><MedlineJournalInfo><Country>United States</Country><MedlineTA>Mol Cell Proteomics</MedlineTA><NlmUniqueID>101125647</NlmUniqueID><ISSNLinking>1535-9476</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D000953">Antigens, Protozoan</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D018985">Epitopes, B-Lymphocyte</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D010455">Peptides</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D000953" MajorTopicYN="N">Antigens, Protozoan</DescriptorName><QualifierName UI="Q000276" MajorTopicYN="N">immunology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D001402" MajorTopicYN="N">B-Lymphocytes</DescriptorName><QualifierName UI="Q000276" MajorTopicYN="N">immunology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D014355" MajorTopicYN="N">Chagas Disease</DescriptorName><QualifierName UI="Q000276" MajorTopicYN="Y">immunology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D030562" MajorTopicYN="N">Databases, Protein</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D004797" MajorTopicYN="N">Enzyme-Linked Immunosorbent Assay</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D018604" MajorTopicYN="N">Epitope Mapping</DescriptorName><QualifierName UI="Q000379" MajorTopicYN="Y">methods</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D018985" MajorTopicYN="N">Epitopes, B-Lymphocyte</DescriptorName><QualifierName UI="Q000276" MajorTopicYN="Y">immunology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D057166" MajorTopicYN="N">High-Throughput Screening Assays</DescriptorName><QualifierName UI="Q000379" MajorTopicYN="Y">methods</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D006801" MajorTopicYN="N">Humans</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D010455" MajorTopicYN="N">Peptides</DescriptorName><QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D040081" MajorTopicYN="N">Protein Array Analysis</DescriptorName><QualifierName UI="Q000379" MajorTopicYN="Y">methods</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D040901" MajorTopicYN="N">Proteomics</DescriptorName><QualifierName UI="Q000379" MajorTopicYN="Y">methods</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D015203" MajorTopicYN="N">Reproducibility of Results</DescriptorName></MeshHeading></MeshHeadingList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="received"><Year>2014</Year><Month>10</Month><Day>27</Day></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2015</Year><Month>4</Month><Day>30</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate 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Carmona</name></noRegion><name sortKey="Aguero, Fernan" sort="Aguero, Fernan" uniqKey="Aguero F" first="Fernán" last="Agüero">Fernán Agüero</name><name sortKey="Altcheh, Jaime" sort="Altcheh, Jaime" uniqKey="Altcheh J" first="Jaime" last="Altcheh">Jaime Altcheh</name><name sortKey="Balouz, Virginia" sort="Balouz, Virginia" uniqKey="Balouz V" first="Virginia" last="Balouz">Virginia Balouz</name><name sortKey="Buscaglia, Carlos A" sort="Buscaglia, Carlos A" uniqKey="Buscaglia C" first="Carlos A" last="Buscaglia">Carlos A. Buscaglia</name><name sortKey="Campetella, Oscar" sort="Campetella, Oscar" uniqKey="Campetella O" first="Oscar" last="Campetella">Oscar Campetella</name><name sortKey="Frasch, Alberto C" sort="Frasch, Alberto C" uniqKey="Frasch A" first="Alberto C" last="Frasch">Alberto C. Frasch</name><name sortKey="Mucci, Juan" sort="Mucci, Juan" uniqKey="Mucci J" first="Juan" last="Mucci">Juan Mucci</name><name sortKey="Tekiel, Valeria" sort="Tekiel, Valeria" uniqKey="Tekiel V" first="Valeria" last="Tekiel">Valeria Tekiel</name></country><country name="Danemark"><noRegion><name sortKey="Nielsen, Morten" sort="Nielsen, Morten" uniqKey="Nielsen M" first="Morten" last="Nielsen">Morten Nielsen</name></noRegion><name sortKey="Schafer Nielsen, Claus" sort="Schafer Nielsen, Claus" uniqKey="Schafer Nielsen C" first="Claus" last="Schafer-Nielsen">Claus Schafer-Nielsen</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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