Characterization of monoclonal antibodies against feline coronavirus accessory protein 7b.
Identifieur interne : 001069 ( Main/Exploration ); précédent : 001068; suivant : 001070Characterization of monoclonal antibodies against feline coronavirus accessory protein 7b.
Auteurs : Tanja Lemmermeyer [Allemagne] ; Benjamin Lamp [Autriche] ; Rainer Schneider [Autriche] ; John Ziebuhr [Allemagne] ; Gergely Tekes [Allemagne] ; Heinz-Jürgen Thiel [Allemagne]Source :
- Veterinary microbiology [ 1873-2542 ] ; 2016.
Descripteurs français
- KwdFr :
- Animaux, Anticorps antiviraux (génétique), Anticorps antiviraux (métabolisme), Anticorps monoclonaux (génétique), Anticorps monoclonaux (métabolisme), Coronavirus félin (immunologie), Escherichia coli (génétique), Glycosylation, Protéines recombinantes (génétique), Protéines recombinantes (immunologie), Protéines virales (génétique), Protéines virales (immunologie), Sites de fixation, Échappement immunitaire.
- MESH :
- génétique : Anticorps antiviraux, Anticorps monoclonaux, Escherichia coli, Protéines recombinantes, Protéines virales.
- immunologie : Coronavirus félin, Protéines recombinantes, Protéines virales.
- métabolisme : Anticorps antiviraux, Anticorps monoclonaux.
- Animaux, Glycosylation, Sites de fixation, Échappement immunitaire.
English descriptors
- KwdEn :
- Animals, Antibodies, Monoclonal (genetics), Antibodies, Monoclonal (metabolism), Antibodies, Viral (genetics), Antibodies, Viral (metabolism), Binding Sites, Coronavirus, Feline (immunology), Escherichia coli (genetics), Glycosylation, Immune Evasion, Recombinant Proteins (genetics), Recombinant Proteins (immunology), Viral Proteins (genetics), Viral Proteins (immunology).
- MESH :
- chemical , genetics : Antibodies, Monoclonal, Antibodies, Viral, Recombinant Proteins, Viral Proteins.
- chemical , immunology : Recombinant Proteins, Viral Proteins.
- chemical , metabolism : Antibodies, Monoclonal, Antibodies, Viral.
- genetics : Escherichia coli.
- immunology : Coronavirus, Feline.
- Animals, Binding Sites, Glycosylation, Immune Evasion.
Abstract
Feline coronaviruses (FCoVs) encode five accessory proteins termed 3a, 3b, 3c, 7a and 7b of unknown function. These proteins are dispensable for viral replication in vitro but are supposed to play a role in virulence. In the current study, we produced and characterized 7b-specific monoclonal antibodies (mAbs). A recombinant form of the 7b protein was expressed as a fusion protein in Escherichia coli, purified by immobilized metal affinity chromatography and used as immunogen. Two hybridoma lines, 5B6 and 14D8, were isolated that expressed mAbs that recognized 7b proteins of both FCoV serotypes. Using an extensive set of N- and C-terminally truncated 7b proteins expressed in E. coli and a synthetic peptide, the binding sites of mAbs 5B6 and 14D8 were mapped to an 18-residue region that comprises the only potential N-glycosylation site of the FCoV 7b protein. The two mAbs were suitable to detect a 24-kDa protein, which represents the nonglycosylated form of 7b in FCoV-infected cells. We speculate that glycosylation of 7b is part of the viral evasion strategy to prevent an immune response against this antigenic site.
DOI: 10.1016/j.vetmic.2015.12.009
PubMed: 26854339
Affiliations:
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Electronic address: heinz-juergen.thiel@vetmed.uni-giessen.de.</nlm:affiliation><country xml:lang="fr">Allemagne</country><wicri:regionArea>Institute of Virology, Justus Liebig University Giessen</wicri:regionArea><wicri:noRegion>Justus Liebig University Giessen</wicri:noRegion><wicri:noRegion>Justus Liebig University Giessen</wicri:noRegion><wicri:noRegion>Justus Liebig University Giessen</wicri:noRegion></affiliation></author></analytic><series><title level="j">Veterinary microbiology</title><idno type="eISSN">1873-2542</idno><imprint><date when="2016" type="published">2016</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term><term>Antibodies, Monoclonal (genetics)</term><term>Antibodies, Monoclonal (metabolism)</term><term>Antibodies, Viral (genetics)</term><term>Antibodies, Viral (metabolism)</term><term>Binding Sites</term><term>Coronavirus, Feline (immunology)</term><term>Escherichia coli (genetics)</term><term>Glycosylation</term><term>Immune Evasion</term><term>Recombinant Proteins (genetics)</term><term>Recombinant Proteins (immunology)</term><term>Viral Proteins (genetics)</term><term>Viral Proteins (immunology)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Animaux</term><term>Anticorps antiviraux (génétique)</term><term>Anticorps antiviraux (métabolisme)</term><term>Anticorps monoclonaux (génétique)</term><term>Anticorps monoclonaux (métabolisme)</term><term>Coronavirus félin (immunologie)</term><term>Escherichia coli (génétique)</term><term>Glycosylation</term><term>Protéines recombinantes (génétique)</term><term>Protéines recombinantes (immunologie)</term><term>Protéines virales (génétique)</term><term>Protéines virales (immunologie)</term><term>Sites de fixation</term><term>Échappement immunitaire</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Antibodies, Monoclonal</term><term>Antibodies, Viral</term><term>Recombinant Proteins</term><term>Viral Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="immunology" xml:lang="en"><term>Recombinant Proteins</term><term>Viral Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Antibodies, Monoclonal</term><term>Antibodies, Viral</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Escherichia coli</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Anticorps antiviraux</term><term>Anticorps monoclonaux</term><term>Escherichia coli</term><term>Protéines recombinantes</term><term>Protéines virales</term></keywords><keywords scheme="MESH" qualifier="immunologie" xml:lang="fr"><term>Coronavirus félin</term><term>Protéines recombinantes</term><term>Protéines virales</term></keywords><keywords scheme="MESH" qualifier="immunology" xml:lang="en"><term>Coronavirus, Feline</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Anticorps antiviraux</term><term>Anticorps monoclonaux</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Animals</term><term>Binding Sites</term><term>Glycosylation</term><term>Immune Evasion</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Animaux</term><term>Glycosylation</term><term>Sites de fixation</term><term>Échappement immunitaire</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Feline coronaviruses (FCoVs) encode five accessory proteins termed 3a, 3b, 3c, 7a and 7b of unknown function. These proteins are dispensable for viral replication in vitro but are supposed to play a role in virulence. In the current study, we produced and characterized 7b-specific monoclonal antibodies (mAbs). A recombinant form of the 7b protein was expressed as a fusion protein in Escherichia coli, purified by immobilized metal affinity chromatography and used as immunogen. Two hybridoma lines, 5B6 and 14D8, were isolated that expressed mAbs that recognized 7b proteins of both FCoV serotypes. Using an extensive set of N- and C-terminally truncated 7b proteins expressed in E. coli and a synthetic peptide, the binding sites of mAbs 5B6 and 14D8 were mapped to an 18-residue region that comprises the only potential N-glycosylation site of the FCoV 7b protein. The two mAbs were suitable to detect a 24-kDa protein, which represents the nonglycosylated form of 7b in FCoV-infected cells. We speculate that glycosylation of 7b is part of the viral evasion strategy to prevent an immune response against this antigenic site. </div></front></TEI><affiliations><list><country><li>Allemagne</li><li>Autriche</li></country><region><li>Vienne (Autriche)</li></region><settlement><li>Vienne (Autriche)</li></settlement></list><tree><country name="Allemagne"><noRegion><name sortKey="Lemmermeyer, Tanja" sort="Lemmermeyer, Tanja" uniqKey="Lemmermeyer T" first="Tanja" last="Lemmermeyer">Tanja Lemmermeyer</name></noRegion><name sortKey="Tekes, Gergely" sort="Tekes, Gergely" uniqKey="Tekes G" first="Gergely" last="Tekes">Gergely Tekes</name><name sortKey="Thiel, Heinz Jurgen" sort="Thiel, Heinz Jurgen" uniqKey="Thiel H" first="Heinz-Jürgen" last="Thiel">Heinz-Jürgen Thiel</name><name sortKey="Ziebuhr, John" sort="Ziebuhr, John" uniqKey="Ziebuhr J" first="John" last="Ziebuhr">John Ziebuhr</name></country><country name="Autriche"><region name="Vienne (Autriche)"><name sortKey="Lamp, Benjamin" sort="Lamp, Benjamin" uniqKey="Lamp B" first="Benjamin" last="Lamp">Benjamin Lamp</name></region><name sortKey="Schneider, Rainer" sort="Schneider, Rainer" uniqKey="Schneider R" first="Rainer" last="Schneider">Rainer Schneider</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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