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Towards High-throughput Immunomics for Infectious Diseases: Use of Next-generation Peptide Microarrays for Rapid Discovery and Mapping of Antigenic Determinants.

Identifieur interne : 001532 ( Main/Exploration ); précédent : 001531; suivant : 001533

Towards 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 :

RBID : pubmed:25922409

Descripteurs français

English descriptors

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


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Le document en format XML

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<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>
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<name sortKey="Balouz, Virginia" sort="Balouz, Virginia" uniqKey="Balouz V" first="Virginia" last="Balouz">Virginia Balouz</name>
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<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>
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<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>
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<name sortKey="Altcheh, Jaime" sort="Altcheh, Jaime" uniqKey="Altcheh J" first="Jaime" last="Altcheh">Jaime Altcheh</name>
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<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>
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<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>
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<name sortKey="Frasch, Alberto C" sort="Frasch, Alberto C" uniqKey="Frasch A" first="Alberto C" last="Frasch">Alberto C. Frasch</name>
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<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>
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<wicri:noRegion>Buenos Aires</wicri:noRegion>
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<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>
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<name sortKey="Buscaglia, Carlos A" sort="Buscaglia, Carlos A" uniqKey="Buscaglia C" first="Carlos A" last="Buscaglia">Carlos A. Buscaglia</name>
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<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>
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<name sortKey="Aguero, Fernan" sort="Aguero, Fernan" uniqKey="Aguero F" first="Fernán" last="Agüero">Fernán Agüero</name>
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<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>
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<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>
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<keywords scheme="MESH" qualifier="immunology" xml:lang="en">
<term>B-Lymphocytes</term>
<term>Chagas Disease</term>
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<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>
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<keywords scheme="MESH" xml:lang="en">
<term>Databases, Protein</term>
<term>Enzyme-Linked Immunosorbent Assay</term>
<term>Humans</term>
<term>Reproducibility of Results</term>
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<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>
<affiliations>
<list>
<country>
<li>Argentine</li>
<li>Danemark</li>
</country>
<region>
<li>Hovedstaden</li>
</region>
<settlement>
<li>Copenhague</li>
</settlement>
</list>
<tree>
<country name="Argentine">
<noRegion>
<name sortKey="Carmona, Santiago J" sort="Carmona, Santiago J" uniqKey="Carmona S" first="Santiago J" last="Carmona">Santiago J. 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>

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