Detection of Epstein-Barr virus-specific memory CD4+ T cells using a peptide-based cultured enzyme-linked immunospot assay.
Identifieur interne : 002016 ( Main/Exploration ); précédent : 002015; suivant : 002017Detection of Epstein-Barr virus-specific memory CD4+ T cells using a peptide-based cultured enzyme-linked immunospot assay.
Auteurs : Sandra A. Calarota [Italie] ; Antonella Chiesa ; Paola Zelini ; Giuditta Comolli ; Lorenzo Minoli ; Fausto BaldantiSource :
- Immunology [ 1365-2567 ] ; 2013.
Descripteurs français
- KwdFr :
- Adulte, Adulte d'âge moyen, Antigènes viraux (immunologie), Cellules cultivées, Femelle, Herpèsvirus humain de type 4 (immunologie), Herpèsvirus humain de type 4 (pathogénicité), Humains, Immunocompétence, Immunosuppresseurs (usage thérapeutique), Infections à virus Epstein-Barr (immunologie), Infections à virus Epstein-Barr (virologie), Interféron gamma (métabolisme), Lymphocytes T CD4+ (immunologie), Lymphocytes T CD4+ (virologie), Marqueurs biologiques (métabolisme), Mâle, Mémoire immunologique, Reproductibilité des résultats, Sujet immunodéprimé, Sujet âgé, Test ELISpot, Tests de libération d'interféron-gamma, Transplantation cardiaque (immunologie), Transplantation pulmonaire (immunologie).
- MESH :
- immunologie : Antigènes viraux, Herpèsvirus humain de type 4, Infections à virus Epstein-Barr, Lymphocytes T CD4+, Transplantation cardiaque, Transplantation pulmonaire.
- métabolisme : Interféron gamma, Marqueurs biologiques.
- pathogénicité : Herpèsvirus humain de type 4.
- usage thérapeutique : Immunosuppresseurs.
- virologie : Infections à virus Epstein-Barr, Lymphocytes T CD4+.
- Adulte, Adulte d'âge moyen, Cellules cultivées, Femelle, Humains, Immunocompétence, Mâle, Mémoire immunologique, Reproductibilité des résultats, Sujet immunodéprimé, Sujet âgé, Test ELISpot, Tests de libération d'interféron-gamma.
English descriptors
- KwdEn :
- Adult, Aged, Antigens, Viral (immunology), Biomarkers (metabolism), CD4-Positive T-Lymphocytes (immunology), CD4-Positive T-Lymphocytes (virology), Cells, Cultured, Enzyme-Linked Immunospot Assay, Epstein-Barr Virus Infections (immunology), Epstein-Barr Virus Infections (virology), Female, Heart Transplantation (immunology), Herpesvirus 4, Human (immunology), Herpesvirus 4, Human (pathogenicity), Humans, Immunocompetence, Immunocompromised Host, Immunologic Memory, Immunosuppressive Agents (therapeutic use), Interferon-gamma (metabolism), Interferon-gamma Release Tests, Lung Transplantation (immunology), Male, Middle Aged, Reproducibility of Results.
- MESH :
- chemical , immunology : Antigens, Viral.
- chemical , metabolism : Biomarkers, Interferon-gamma.
- immunology : CD4-Positive T-Lymphocytes, Epstein-Barr Virus Infections, Heart Transplantation, Herpesvirus 4, Human, Lung Transplantation.
- pathogenicity : Herpesvirus 4, Human.
- chemical , therapeutic use : Immunosuppressive Agents.
- virology : CD4-Positive T-Lymphocytes, Epstein-Barr Virus Infections.
- Adult, Aged, Cells, Cultured, Enzyme-Linked Immunospot Assay, Female, Humans, Immunocompetence, Immunocompromised Host, Immunologic Memory, Interferon-gamma Release Tests, Male, Middle Aged, Reproducibility of Results.
Abstract
Approaches to evaluate T-cell responses to Epstein-Barr virus (EBV) include enzyme-linked immunospot (ELISPOT), which quantifies cells capable of immediate interferon-γ secretion upon antigen stimulation. However, evaluation of expandable EBV-specific memory T cells in an ELISPOT format has not been described previously. We quantified EBV-specific T-cell precursors with high proliferative capacity by using a peptide-based cultured interferon-γ ELISPOT assay. Standard and cultured ELISPOT responses to overlapping peptide pools (15-mers overlapping by 11 amino acids) covering the lytic (BZLF1 and BMRF1) and latent (EBNA1, EBNA3a, EBNA3b, EBNA3c, LMP1 and LMP2) EBV proteins were evaluated in 20 healthy subjects with remote EBV infection and, for comparison, in four solid organ transplant recipients. Cultured ELISPOT responses to both lytic and latent EBV antigens were significantly higher than standard ELISPOT responses. The distribution of EBV-specific T-cell responses detected in healthy virus carriers showed more consistent cultured ELISPOT responses compared with standard ELISPOT responses. T-cell responses quantified by cultured ELISPOT were mainly mediated by CD4+ T cells and a marked pattern of immunodominance to latent-phase antigens (EBNA1 > EBNA3 family antigens > LMP2 > LMP1) was shown. Both the magnitude and distribution of EBV-specific T-cell responses were altered in solid organ transplant recipients; in particular, cultured ELISPOT responses were almost undetectable in a lung-transplanted patient with EBV-associated diseases. Analysis of T-cell responses to EBV by ELISPOT assays might provide new insights into the pathogenesis of EBV-related diseases and serve as new tools in the monitoring of EBV infection in immunocompromised patients.
DOI: 10.1111/imm.12106
PubMed: 23560877
Affiliations:
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Le document en format XML
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Calarota</name><affiliation wicri:level="1"><nlm:affiliation>Virology and Microbiology Department, Fondazione IRCCS Policlinico San Matteo, Pavia, Italy.</nlm:affiliation><country xml:lang="fr">Italie</country><wicri:regionArea>Virology and Microbiology Department, Fondazione IRCCS Policlinico San Matteo, Pavia</wicri:regionArea><wicri:noRegion>Pavia</wicri:noRegion></affiliation></author><author><name sortKey="Chiesa, Antonella" sort="Chiesa, Antonella" uniqKey="Chiesa A" first="Antonella" last="Chiesa">Antonella Chiesa</name></author><author><name sortKey="Zelini, Paola" sort="Zelini, Paola" uniqKey="Zelini P" first="Paola" last="Zelini">Paola Zelini</name></author><author><name sortKey="Comolli, Giuditta" sort="Comolli, Giuditta" uniqKey="Comolli G" first="Giuditta" last="Comolli">Giuditta Comolli</name></author><author><name sortKey="Minoli, Lorenzo" sort="Minoli, Lorenzo" uniqKey="Minoli L" first="Lorenzo" last="Minoli">Lorenzo Minoli</name></author><author><name sortKey="Baldanti, Fausto" sort="Baldanti, Fausto" uniqKey="Baldanti F" first="Fausto" last="Baldanti">Fausto Baldanti</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2013">2013</date><idno type="RBID">pubmed:23560877</idno><idno type="pmid">23560877</idno><idno type="doi">10.1111/imm.12106</idno><idno type="wicri:Area/PubMed/Corpus">001D05</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">001D05</idno><idno type="wicri:Area/PubMed/Curation">001D05</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Curation">001D05</idno><idno type="wicri:Area/PubMed/Checkpoint">001B67</idno><idno type="wicri:explorRef" wicri:stream="Checkpoint" wicri:step="PubMed">001B67</idno><idno type="wicri:Area/Ncbi/Merge">000A35</idno><idno type="wicri:Area/Ncbi/Curation">000A35</idno><idno type="wicri:Area/Ncbi/Checkpoint">000A35</idno><idno type="wicri:Area/Main/Merge">002037</idno><idno type="wicri:Area/Main/Curation">002016</idno><idno type="wicri:Area/Main/Exploration">002016</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Detection of Epstein-Barr virus-specific memory CD4+ T cells using a peptide-based cultured enzyme-linked immunospot assay.</title><author><name sortKey="Calarota, Sandra A" sort="Calarota, Sandra A" uniqKey="Calarota S" first="Sandra A" last="Calarota">Sandra A. 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(pathogenicity)</term><term>Humans</term><term>Immunocompetence</term><term>Immunocompromised Host</term><term>Immunologic Memory</term><term>Immunosuppressive Agents (therapeutic use)</term><term>Interferon-gamma (metabolism)</term><term>Interferon-gamma Release Tests</term><term>Lung Transplantation (immunology)</term><term>Male</term><term>Middle Aged</term><term>Reproducibility of Results</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Adulte</term><term>Adulte d'âge moyen</term><term>Antigènes viraux (immunologie)</term><term>Cellules cultivées</term><term>Femelle</term><term>Herpèsvirus humain de type 4 (immunologie)</term><term>Herpèsvirus humain de type 4 (pathogénicité)</term><term>Humains</term><term>Immunocompétence</term><term>Immunosuppresseurs (usage thérapeutique)</term><term>Infections à virus Epstein-Barr (immunologie)</term><term>Infections à virus Epstein-Barr (virologie)</term><term>Interféron gamma (métabolisme)</term><term>Lymphocytes T CD4+ (immunologie)</term><term>Lymphocytes T CD4+ (virologie)</term><term>Marqueurs biologiques (métabolisme)</term><term>Mâle</term><term>Mémoire immunologique</term><term>Reproductibilité des résultats</term><term>Sujet immunodéprimé</term><term>Sujet âgé</term><term>Test ELISpot</term><term>Tests de libération d'interféron-gamma</term><term>Transplantation cardiaque (immunologie)</term><term>Transplantation pulmonaire (immunologie)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="immunology" xml:lang="en"><term>Antigens, Viral</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Biomarkers</term><term>Interferon-gamma</term></keywords><keywords scheme="MESH" qualifier="immunologie" xml:lang="fr"><term>Antigènes viraux</term><term>Herpèsvirus humain de type 4</term><term>Infections à virus Epstein-Barr</term><term>Lymphocytes T CD4+</term><term>Transplantation cardiaque</term><term>Transplantation 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Epstein-Barr</term><term>Lymphocytes T CD4+</term></keywords><keywords scheme="MESH" qualifier="virology" xml:lang="en"><term>CD4-Positive T-Lymphocytes</term><term>Epstein-Barr Virus Infections</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Adult</term><term>Aged</term><term>Cells, Cultured</term><term>Enzyme-Linked Immunospot Assay</term><term>Female</term><term>Humans</term><term>Immunocompetence</term><term>Immunocompromised Host</term><term>Immunologic Memory</term><term>Interferon-gamma Release Tests</term><term>Male</term><term>Middle Aged</term><term>Reproducibility of Results</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Adulte</term><term>Adulte d'âge moyen</term><term>Cellules cultivées</term><term>Femelle</term><term>Humains</term><term>Immunocompétence</term><term>Mâle</term><term>Mémoire immunologique</term><term>Reproductibilité des résultats</term><term>Sujet immunodéprimé</term><term>Sujet âgé</term><term>Test ELISpot</term><term>Tests de libération d'interféron-gamma</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Approaches to evaluate T-cell responses to Epstein-Barr virus (EBV) include enzyme-linked immunospot (ELISPOT), which quantifies cells capable of immediate interferon-γ secretion upon antigen stimulation. However, evaluation of expandable EBV-specific memory T cells in an ELISPOT format has not been described previously. We quantified EBV-specific T-cell precursors with high proliferative capacity by using a peptide-based cultured interferon-γ ELISPOT assay. Standard and cultured ELISPOT responses to overlapping peptide pools (15-mers overlapping by 11 amino acids) covering the lytic (BZLF1 and BMRF1) and latent (EBNA1, EBNA3a, EBNA3b, EBNA3c, LMP1 and LMP2) EBV proteins were evaluated in 20 healthy subjects with remote EBV infection and, for comparison, in four solid organ transplant recipients. Cultured ELISPOT responses to both lytic and latent EBV antigens were significantly higher than standard ELISPOT responses. The distribution of EBV-specific T-cell responses detected in healthy virus carriers showed more consistent cultured ELISPOT responses compared with standard ELISPOT responses. T-cell responses quantified by cultured ELISPOT were mainly mediated by CD4+ T cells and a marked pattern of immunodominance to latent-phase antigens (EBNA1 > EBNA3 family antigens > LMP2 > LMP1) was shown. Both the magnitude and distribution of EBV-specific T-cell responses were altered in solid organ transplant recipients; in particular, cultured ELISPOT responses were almost undetectable in a lung-transplanted patient with EBV-associated diseases. Analysis of T-cell responses to EBV by ELISPOT assays might provide new insights into the pathogenesis of EBV-related diseases and serve as new tools in the monitoring of EBV infection in immunocompromised patients.</div></front></TEI><affiliations><list><country><li>Italie</li></country></list><tree><noCountry><name sortKey="Baldanti, Fausto" sort="Baldanti, Fausto" uniqKey="Baldanti F" first="Fausto" last="Baldanti">Fausto Baldanti</name><name sortKey="Chiesa, Antonella" sort="Chiesa, Antonella" uniqKey="Chiesa A" first="Antonella" last="Chiesa">Antonella Chiesa</name><name sortKey="Comolli, Giuditta" sort="Comolli, Giuditta" uniqKey="Comolli G" first="Giuditta" last="Comolli">Giuditta Comolli</name><name sortKey="Minoli, Lorenzo" sort="Minoli, Lorenzo" uniqKey="Minoli L" first="Lorenzo" last="Minoli">Lorenzo Minoli</name><name sortKey="Zelini, Paola" sort="Zelini, Paola" uniqKey="Zelini P" first="Paola" last="Zelini">Paola Zelini</name></noCountry><country name="Italie"><noRegion><name sortKey="Calarota, Sandra A" sort="Calarota, Sandra A" uniqKey="Calarota S" first="Sandra A" last="Calarota">Sandra A. 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