Contamination of synthetic HuD protein spanning peptide pools with a CMV-encoded peptide.
Identifieur interne : 002997 ( Main/Exploration ); précédent : 002996; suivant : 002998Contamination of synthetic HuD protein spanning peptide pools with a CMV-encoded peptide.
Auteurs : Marieke T. De Graaf [Pays-Bas] ; Janet W. De Beukelaar ; Peter C. Burgers ; Theo M. Luider ; Jaco Kraan ; Peter A Sillevis Smitt ; Jan W. GratamaSource :
- Cytometry. Part A : the journal of the International Society for Analytical Cytology [ 1552-4930 ] ; 2008.
Descripteurs français
- KwdFr :
- Activation des lymphocytes, Antigènes Hu de l'encéphalomyélite paranéoplasique (), Antigènes Hu de l'encéphalomyélite paranéoplasique (immunologie), Cytomegalovirus (immunologie), Données de séquences moléculaires, Humains, Lymphocytes T CD8+ (immunologie), Peptides (), Peptides (immunologie), Spectrométrie de masse MALDI, Séquence d'acides aminés, Test d'histocompatibilité.
- MESH :
- immunologie : Antigènes Hu de l'encéphalomyélite paranéoplasique, Cytomegalovirus, Lymphocytes T CD8+, Peptides.
- Activation des lymphocytes, Antigènes Hu de l'encéphalomyélite paranéoplasique, Données de séquences moléculaires, Humains, Peptides, Spectrométrie de masse MALDI, Séquence d'acides aminés, Test d'histocompatibilité.
English descriptors
- KwdEn :
- Amino Acid Sequence, CD8-Positive T-Lymphocytes (immunology), Cytomegalovirus (immunology), ELAV Proteins (chemistry), ELAV Proteins (immunology), Histocompatibility Testing, Humans, Lymphocyte Activation, Molecular Sequence Data, Peptides (chemistry), Peptides (immunology), Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization.
- MESH :
- chemical , chemistry : ELAV Proteins, Peptides.
- immunology : CD8-Positive T-Lymphocytes, Cytomegalovirus, ELAV Proteins, Peptides.
- Amino Acid Sequence, Histocompatibility Testing, Humans, Lymphocyte Activation, Molecular Sequence Data, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization.
Abstract
To detect HuD-specific T cells in patients with Hu-antibody associated paraneoplastic neurological syndromes (Hu-PNS), we used short-term stimulation assays with HuD protein spanning peptide pools (PSPP) with purities of at least 70% and found reproducible false-positive CD8+ T-cell responses in three of 127 individuals (two healthy controls and one Hu-PNS patient), which all shared HLA-A*2402 and HLA-B*1801. After testing the 15-mer peptides of the HuD antigen separately, we discovered that the same three 15-mers yielded the CD8+ T cell response in those three individuals. This highly unusual result could not be reproduced when using new batches of peptides with a higher level of purity (>82% and >95%). Therefore, we assumed this response was not directed against the HuD peptides and analyzed the HuD 15-mers by Fourier transform ion cyclotron resonance (FT-ICR) tandem mass spectrometry (MS/MS), which showed the presence of a cytomegalovirus (CMV)-encoded peptide (AIAEESDEEEAIVAY) as a contaminant. The three responding individuals all were CMV-seropositive and the contaminating peptide appeared to fit in the binding groove of HLA-B*18. Our data reveal that synthetic PSPP may contain immunogenic contaminations which may cause false positive results in T-cell stimulation assays.
DOI: 10.1002/cyto.a.20636
PubMed: 18727061
Affiliations:
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Le document en format XML
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De Graaf</name><affiliation wicri:level="3"><nlm:affiliation>Department of Neurology, Erasmus University Medical Center, Rotterdam, The Netherlands.</nlm:affiliation><country xml:lang="fr">Pays-Bas</country><wicri:regionArea>Department of Neurology, Erasmus University Medical Center, Rotterdam</wicri:regionArea><placeName><settlement type="city">Rotterdam</settlement><region nuts="2" type="province">Hollande-Méridionale</region></placeName></affiliation></author><author><name sortKey="De Beukelaar, Janet W" sort="De Beukelaar, Janet W" uniqKey="De Beukelaar J" first="Janet W" last="De Beukelaar">Janet W. De Beukelaar</name></author><author><name sortKey="Burgers, Peter C" sort="Burgers, Peter C" uniqKey="Burgers P" first="Peter C" last="Burgers">Peter C. Burgers</name></author><author><name sortKey="Luider, Theo M" sort="Luider, Theo M" uniqKey="Luider T" first="Theo M" last="Luider">Theo M. Luider</name></author><author><name sortKey="Kraan, Jaco" sort="Kraan, Jaco" uniqKey="Kraan J" first="Jaco" last="Kraan">Jaco Kraan</name></author><author><name sortKey="Smitt, Peter A Sillevis" sort="Smitt, Peter A Sillevis" uniqKey="Smitt P" first="Peter A Sillevis" last="Smitt">Peter A Sillevis Smitt</name></author><author><name sortKey="Gratama, Jan W" sort="Gratama, Jan W" uniqKey="Gratama J" first="Jan W" last="Gratama">Jan W. Gratama</name></author></analytic><series><title level="j">Cytometry. Part A : the journal of the International Society for Analytical Cytology</title><idno type="eISSN">1552-4930</idno><imprint><date when="2008" type="published">2008</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Amino Acid Sequence</term><term>CD8-Positive T-Lymphocytes (immunology)</term><term>Cytomegalovirus (immunology)</term><term>ELAV Proteins (chemistry)</term><term>ELAV Proteins (immunology)</term><term>Histocompatibility Testing</term><term>Humans</term><term>Lymphocyte Activation</term><term>Molecular Sequence Data</term><term>Peptides (chemistry)</term><term>Peptides (immunology)</term><term>Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Activation des lymphocytes</term><term>Antigènes Hu de l'encéphalomyélite paranéoplasique ()</term><term>Antigènes Hu de l'encéphalomyélite paranéoplasique (immunologie)</term><term>Cytomegalovirus (immunologie)</term><term>Données de séquences moléculaires</term><term>Humains</term><term>Lymphocytes T CD8+ (immunologie)</term><term>Peptides ()</term><term>Peptides (immunologie)</term><term>Spectrométrie de masse MALDI</term><term>Séquence d'acides aminés</term><term>Test d'histocompatibilité</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>ELAV Proteins</term><term>Peptides</term></keywords><keywords scheme="MESH" qualifier="immunologie" xml:lang="fr"><term>Antigènes Hu de l'encéphalomyélite paranéoplasique</term><term>Cytomegalovirus</term><term>Lymphocytes T CD8+</term><term>Peptides</term></keywords><keywords scheme="MESH" qualifier="immunology" xml:lang="en"><term>CD8-Positive T-Lymphocytes</term><term>Cytomegalovirus</term><term>ELAV Proteins</term><term>Peptides</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Amino Acid Sequence</term><term>Histocompatibility Testing</term><term>Humans</term><term>Lymphocyte Activation</term><term>Molecular Sequence Data</term><term>Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Activation des lymphocytes</term><term>Antigènes Hu de l'encéphalomyélite paranéoplasique</term><term>Données de séquences moléculaires</term><term>Humains</term><term>Peptides</term><term>Spectrométrie de masse MALDI</term><term>Séquence d'acides aminés</term><term>Test d'histocompatibilité</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">To detect HuD-specific T cells in patients with Hu-antibody associated paraneoplastic neurological syndromes (Hu-PNS), we used short-term stimulation assays with HuD protein spanning peptide pools (PSPP) with purities of at least 70% and found reproducible false-positive CD8+ T-cell responses in three of 127 individuals (two healthy controls and one Hu-PNS patient), which all shared HLA-A*2402 and HLA-B*1801. After testing the 15-mer peptides of the HuD antigen separately, we discovered that the same three 15-mers yielded the CD8+ T cell response in those three individuals. This highly unusual result could not be reproduced when using new batches of peptides with a higher level of purity (>82% and >95%). Therefore, we assumed this response was not directed against the HuD peptides and analyzed the HuD 15-mers by Fourier transform ion cyclotron resonance (FT-ICR) tandem mass spectrometry (MS/MS), which showed the presence of a cytomegalovirus (CMV)-encoded peptide (AIAEESDEEEAIVAY) as a contaminant. The three responding individuals all were CMV-seropositive and the contaminating peptide appeared to fit in the binding groove of HLA-B*18. Our data reveal that synthetic PSPP may contain immunogenic contaminations which may cause false positive results in T-cell stimulation assays.</div></front></TEI><affiliations><list><country><li>Pays-Bas</li></country><region><li>Hollande-Méridionale</li></region><settlement><li>Rotterdam</li></settlement></list><tree><noCountry><name sortKey="Burgers, Peter C" sort="Burgers, Peter C" uniqKey="Burgers P" first="Peter C" last="Burgers">Peter C. Burgers</name><name sortKey="De Beukelaar, Janet W" sort="De Beukelaar, Janet W" uniqKey="De Beukelaar J" first="Janet W" last="De Beukelaar">Janet W. De Beukelaar</name><name sortKey="Gratama, Jan W" sort="Gratama, Jan W" uniqKey="Gratama J" first="Jan W" last="Gratama">Jan W. Gratama</name><name sortKey="Kraan, Jaco" sort="Kraan, Jaco" uniqKey="Kraan J" first="Jaco" last="Kraan">Jaco Kraan</name><name sortKey="Luider, Theo M" sort="Luider, Theo M" uniqKey="Luider T" first="Theo M" last="Luider">Theo M. Luider</name><name sortKey="Smitt, Peter A Sillevis" sort="Smitt, Peter A Sillevis" uniqKey="Smitt P" first="Peter A Sillevis" last="Smitt">Peter A Sillevis Smitt</name></noCountry><country name="Pays-Bas"><region name="Hollande-Méridionale"><name sortKey="De Graaf, Marieke T" sort="De Graaf, Marieke T" uniqKey="De Graaf M" first="Marieke T" last="De Graaf">Marieke T. De Graaf</name></region></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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