Transcriptome profiling of gene expression during immunisation trial against Fasciola hepatica: identification of genes and pathways involved in conferring immunoprotection in a murine model

Fasciolosis remains a significant food-borne trematode disease causing high morbidity around the world and affecting grazing animals and humans. A deeper understanding concerning the molecular mechanisms by which Fasciola hepatica infection occurs, as well as the molecular basis involved in acquirin...

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Veröffentlicht in:BMC infectious diseases 2017-01, Vol.17 (1), p.94-94, Article 94
Hauptverfasser: Rojas-Caraballo, Jose, López-Abán, Julio, Moreno-Pérez, Darwin Andrés, Vicente, Belén, Fernández-Soto, Pedro, Del Olmo, Esther, Patarroyo, Manuel Alfonso, Muro, Antonio
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container_end_page 94
container_issue 1
container_start_page 94
container_title BMC infectious diseases
container_volume 17
creator Rojas-Caraballo, Jose
López-Abán, Julio
Moreno-Pérez, Darwin Andrés
Vicente, Belén
Fernández-Soto, Pedro
Del Olmo, Esther
Patarroyo, Manuel Alfonso
Muro, Antonio
description Fasciolosis remains a significant food-borne trematode disease causing high morbidity around the world and affecting grazing animals and humans. A deeper understanding concerning the molecular mechanisms by which Fasciola hepatica infection occurs, as well as the molecular basis involved in acquiring protection is extremely important when designing and selecting new vaccine candidates. The present study provides a first report of microarray-based technology for describing changes in the splenic gene expression profile for mice immunised with a highly effective, protection-inducing, multi-epitope, subunit-based, chemically-synthesised vaccine candidate against F. hepatica. The mice were immunised with synthetic peptides containing B- and T-cell epitopes, which are derived from F. hepatica cathepsin B and amoebapore proteins, as novel vaccine candidates against F. hepatica formulated in an adjuvant adaptation vaccination system; they were experimentally challenged with F. hepatica metacercariae. Spleen RNA from mice immunised with the highest protection-inducing synthetic peptides was isolated, amplified and labelled using Affymetrix standardised protocols. Data was then background corrected, normalised and the expression signal was calculated. The Ingenuity Pathway Analysis tool was then used for analysing differentially expressed gene identifiers for annotating bio-functions and constructing and visualising molecular interaction networks. Mice immunised with a combination of three peptides containing T-cell epitopes induced high protection against experimental challenge according to survival rates and hepatic damage scores. It also induced differential expression of 820 genes, 168 genes being up-regulated and 652 genes being down-regulated, p value
doi_str_mv 10.1186/s12879-017-2205-3
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A deeper understanding concerning the molecular mechanisms by which Fasciola hepatica infection occurs, as well as the molecular basis involved in acquiring protection is extremely important when designing and selecting new vaccine candidates. The present study provides a first report of microarray-based technology for describing changes in the splenic gene expression profile for mice immunised with a highly effective, protection-inducing, multi-epitope, subunit-based, chemically-synthesised vaccine candidate against F. hepatica. The mice were immunised with synthetic peptides containing B- and T-cell epitopes, which are derived from F. hepatica cathepsin B and amoebapore proteins, as novel vaccine candidates against F. hepatica formulated in an adjuvant adaptation vaccination system; they were experimentally challenged with F. hepatica metacercariae. Spleen RNA from mice immunised with the highest protection-inducing synthetic peptides was isolated, amplified and labelled using Affymetrix standardised protocols. Data was then background corrected, normalised and the expression signal was calculated. The Ingenuity Pathway Analysis tool was then used for analysing differentially expressed gene identifiers for annotating bio-functions and constructing and visualising molecular interaction networks. Mice immunised with a combination of three peptides containing T-cell epitopes induced high protection against experimental challenge according to survival rates and hepatic damage scores. It also induced differential expression of 820 genes, 168 genes being up-regulated and 652 genes being down-regulated, p value &lt;0.05, fold change ranging from -2.944 to 7.632. A functional study of these genes revealed changes in the pathways related to nitric oxide and reactive oxygen species production, Interleukin-12 signalling and production in macrophages and Interleukin-8 signalling with up-regulation of S100 calcium-binding protein A8, Matrix metallopeptidase 9 and CXC chemokine receptor 2 genes. 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A deeper understanding concerning the molecular mechanisms by which Fasciola hepatica infection occurs, as well as the molecular basis involved in acquiring protection is extremely important when designing and selecting new vaccine candidates. The present study provides a first report of microarray-based technology for describing changes in the splenic gene expression profile for mice immunised with a highly effective, protection-inducing, multi-epitope, subunit-based, chemically-synthesised vaccine candidate against F. hepatica. The mice were immunised with synthetic peptides containing B- and T-cell epitopes, which are derived from F. hepatica cathepsin B and amoebapore proteins, as novel vaccine candidates against F. hepatica formulated in an adjuvant adaptation vaccination system; they were experimentally challenged with F. hepatica metacercariae. Spleen RNA from mice immunised with the highest protection-inducing synthetic peptides was isolated, amplified and labelled using Affymetrix standardised protocols. Data was then background corrected, normalised and the expression signal was calculated. The Ingenuity Pathway Analysis tool was then used for analysing differentially expressed gene identifiers for annotating bio-functions and constructing and visualising molecular interaction networks. Mice immunised with a combination of three peptides containing T-cell epitopes induced high protection against experimental challenge according to survival rates and hepatic damage scores. It also induced differential expression of 820 genes, 168 genes being up-regulated and 652 genes being down-regulated, p value &lt;0.05, fold change ranging from -2.944 to 7.632. A functional study of these genes revealed changes in the pathways related to nitric oxide and reactive oxygen species production, Interleukin-12 signalling and production in macrophages and Interleukin-8 signalling with up-regulation of S100 calcium-binding protein A8, Matrix metallopeptidase 9 and CXC chemokine receptor 2 genes. The data obtained in the present study provided us with a more comprehensive overview concerning the possible molecular pathways implied in inducing protection against F. hepatica in a murine model, which could be useful for evaluating future vaccine candidates.</description><subject>Amino acids</subject><subject>Animals</subject><subject>Antibodies, Helminth - immunology</subject><subject>Antigens</subject><subject>Calgranulin A - drug effects</subject><subject>Calgranulin A - genetics</subject><subject>Care and treatment</subject><subject>Development and progression</subject><subject>Disease</subject><subject>Epitopes - immunology</subject><subject>Fasciola hepatica - immunology</subject><subject>Fascioliasis - prevention &amp; control</subject><subject>Female</subject><subject>Fluke infections</subject><subject>Gene expression</subject><subject>Gene Expression - drug effects</subject><subject>Gene Expression Profiling</subject><subject>Genetic aspects</subject><subject>Health aspects</subject><subject>Immune response</subject><subject>Infections</subject><subject>Interleukin-12 - genetics</subject><subject>Interleukin-8 - drug effects</subject><subject>Interleukin-8 - genetics</subject><subject>Laboratory animals</subject><subject>Matrix Metalloproteinase 9 - drug effects</subject><subject>Matrix Metalloproteinase 9 - genetics</subject><subject>Mice</subject><subject>Parasites</subject><subject>Peptides</subject><subject>Peptides - immunology</subject><subject>Proteins</subject><subject>Protozoan Vaccines - pharmacology</subject><subject>Receptors, Interleukin-8B - drug effects</subject><subject>Receptors, Interleukin-8B - genetics</subject><subject>RNA, Messenger - drug effects</subject><subject>RNA, Messenger - metabolism</subject><subject>Spleen - drug effects</subject><subject>Spleen - metabolism</subject><subject>Testing</subject><subject>Up-Regulation</subject><subject>Vaccination</subject><subject>Vaccines</subject><issn>1471-2334</issn><issn>1471-2334</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2017</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>BENPR</sourceid><recordid>eNqNks9u1DAQxiMEoqXwAFyQJS5wSPGfZO3lUKmqKFSqVAkKV2vWnqSuEnuxk6V9OZ4NZ3cpu4gDsiKPMr_vG3s8RfGS0WPG1OxdYlzJeUmZLDmndSkeFYeskqzkQlSPd-KD4llKtzSDis-fFgdcMVYppQ6Ln9cRfDLRLYfQI1nG0LjO-ZaEhrTokeDdMmJKLnhixzhlXN-P3iUYpn9DdNARaMH5NJBzSMaFDsgNLnPewHviLPrBNTle81vbRMBbkpmbH3CfiPOr0K3Q5oCY4BuMfyqFfKYBzVqd00D66RhI-mCxe148aaBL-GK7HxVfzz9cn30qL68-XpydXpamnvGhxBlKgwJAgZoLTithKMi6UZbbheWMGWkYp0bmbyEYF7ioGmmslaapLZXiqDjZ-C7HRY_W5DtF6PQyuh7ivQ7g9H7GuxvdhpWueT1XNc8Gb7YGMXwfMQ26d8lg14HHMCadn5PNKJ-va73-C70NY_T5ehMlJKWq3qFa6FA734Rc10ym-rRSdEZrJifq-B9UXhZ7lzuN-bVxX_B2T5CZAe-GFsaU9MWXz__PXn3bZ9mGNTGkFLF56B2jehpmvRlmnWdUT8OsRda82m36g-L39Ipfl77zqQ</recordid><startdate>20170123</startdate><enddate>20170123</enddate><creator>Rojas-Caraballo, Jose</creator><creator>López-Abán, Julio</creator><creator>Moreno-Pérez, Darwin Andrés</creator><creator>Vicente, Belén</creator><creator>Fernández-Soto, Pedro</creator><creator>Del Olmo, Esther</creator><creator>Patarroyo, Manuel Alfonso</creator><creator>Muro, Antonio</creator><general>BioMed Central Ltd</general><general>BioMed Central</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>IOV</scope><scope>ISR</scope><scope>3V.</scope><scope>7QL</scope><scope>7T2</scope><scope>7U9</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8C1</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BENPR</scope><scope>C1K</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>H94</scope><scope>K9.</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>20170123</creationdate><title>Transcriptome profiling of gene expression during immunisation trial against Fasciola hepatica: identification of genes and pathways involved in conferring immunoprotection in a murine model</title><author>Rojas-Caraballo, Jose ; 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A deeper understanding concerning the molecular mechanisms by which Fasciola hepatica infection occurs, as well as the molecular basis involved in acquiring protection is extremely important when designing and selecting new vaccine candidates. The present study provides a first report of microarray-based technology for describing changes in the splenic gene expression profile for mice immunised with a highly effective, protection-inducing, multi-epitope, subunit-based, chemically-synthesised vaccine candidate against F. hepatica. The mice were immunised with synthetic peptides containing B- and T-cell epitopes, which are derived from F. hepatica cathepsin B and amoebapore proteins, as novel vaccine candidates against F. hepatica formulated in an adjuvant adaptation vaccination system; they were experimentally challenged with F. hepatica metacercariae. Spleen RNA from mice immunised with the highest protection-inducing synthetic peptides was isolated, amplified and labelled using Affymetrix standardised protocols. Data was then background corrected, normalised and the expression signal was calculated. The Ingenuity Pathway Analysis tool was then used for analysing differentially expressed gene identifiers for annotating bio-functions and constructing and visualising molecular interaction networks. Mice immunised with a combination of three peptides containing T-cell epitopes induced high protection against experimental challenge according to survival rates and hepatic damage scores. It also induced differential expression of 820 genes, 168 genes being up-regulated and 652 genes being down-regulated, p value &lt;0.05, fold change ranging from -2.944 to 7.632. A functional study of these genes revealed changes in the pathways related to nitric oxide and reactive oxygen species production, Interleukin-12 signalling and production in macrophages and Interleukin-8 signalling with up-regulation of S100 calcium-binding protein A8, Matrix metallopeptidase 9 and CXC chemokine receptor 2 genes. The data obtained in the present study provided us with a more comprehensive overview concerning the possible molecular pathways implied in inducing protection against F. hepatica in a murine model, which could be useful for evaluating future vaccine candidates.</abstract><cop>England</cop><pub>BioMed Central Ltd</pub><pmid>28114888</pmid><doi>10.1186/s12879-017-2205-3</doi><tpages>1</tpages><oa>free_for_read</oa></addata></record>
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subjects Amino acids
Animals
Antibodies, Helminth - immunology
Antigens
Calgranulin A - drug effects
Calgranulin A - genetics
Care and treatment
Development and progression
Disease
Epitopes - immunology
Fasciola hepatica - immunology
Fascioliasis - prevention & control
Female
Fluke infections
Gene expression
Gene Expression - drug effects
Gene Expression Profiling
Genetic aspects
Health aspects
Immune response
Infections
Interleukin-12 - genetics
Interleukin-8 - drug effects
Interleukin-8 - genetics
Laboratory animals
Matrix Metalloproteinase 9 - drug effects
Matrix Metalloproteinase 9 - genetics
Mice
Parasites
Peptides
Peptides - immunology
Proteins
Protozoan Vaccines - pharmacology
Receptors, Interleukin-8B - drug effects
Receptors, Interleukin-8B - genetics
RNA, Messenger - drug effects
RNA, Messenger - metabolism
Spleen - drug effects
Spleen - metabolism
Testing
Up-Regulation
Vaccination
Vaccines
title Transcriptome profiling of gene expression during immunisation trial against Fasciola hepatica: identification of genes and pathways involved in conferring immunoprotection in a murine model
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