MersV1, Main, Exploration, bibRecord, 003661

Differential antigenicity of recombinant polyepitope-antigens based on loop- and helix-forming B and T cell epitopes

Identifieur interne : 003661 ( Main/Exploration ); précédent : 003660; suivant : 003662

Differential antigenicity of recombinant polyepitope-antigens based on loop- and helix-forming B and T cell epitopes

Auteurs : D. M Theisen [Luxembourg (pays)] ; F. B Bouche [Luxembourg (pays)] ; K. C El Kasmi [Luxembourg (pays)] ; I. Von Der Ahe [Luxembourg (pays)] ; W. Ammerlaan [Luxembourg (pays)] ; S. Demotz [Suisse] ; C. P Muller [Luxembourg (pays)]

Source :

Journal of Immunological Methods [ 0022-1759 ] ; 2000.

RBID : ISTEX:15E2F4832F500C24C588C078CCDE7CA309CBA52D

Descripteurs français

English descriptors

KwdEn :
- Amino Acid Sequence, Animals, Antigen, Antigenic determinant, Antigenicity, Antigens, Viral (biosynthesis), Antigens, Viral (chemistry), Antigens, Viral (genetics), Antigens, Viral (immunology), B-Lymphocyte, Cell Line, Chimera, Cricetinae, Epitopes, B-Lymphocyte (biosynthesis), Epitopes, B-Lymphocyte (chemistry), Epitopes, B-Lymphocyte (genetics), Epitopes, B-Lymphocyte (immunology), Epitopes, T-Lymphocyte (biosynthesis), Epitopes, T-Lymphocyte (chemistry), Epitopes, T-Lymphocyte (genetics), Epitopes, T-Lymphocyte (immunology), Gene Expression, Helical structure, Hemagglutinin, Hemagglutinins, Viral (biosynthesis), Hemagglutinins, Viral (chemistry), Hemagglutinins, Viral (genetics), Hemagglutinins, Viral (immunology), Measles virus, Molecular Sequence Data, Peptides (chemistry), Peptides (genetics), Peptides (immunology), Protein, Protein Conformation, Recombinant Fusion Proteins (chemistry), Recombinant Fusion Proteins (genetics), Recombinant Fusion Proteins (immunology), T-Lymphocyte.
MESH :
- chemical , biosynthesis : Antigens, Viral, Epitopes, B-Lymphocyte, Epitopes, T-Lymphocyte, Hemagglutinins, Viral.
- chemical , chemistry : Antigens, Viral, Epitopes, B-Lymphocyte, Epitopes, T-Lymphocyte, Hemagglutinins, Viral, Peptides, Recombinant Fusion Proteins.
- chemical , genetics : Antigens, Viral, Epitopes, B-Lymphocyte, Epitopes, T-Lymphocyte, Hemagglutinins, Viral, Peptides, Recombinant Fusion Proteins.
- chemical , immunology : Antigens, Viral, Epitopes, B-Lymphocyte, Epitopes, T-Lymphocyte, Hemagglutinins, Viral, Peptides, Recombinant Fusion Proteins.
- Amino Acid Sequence, Animals, Cell Line, Cricetinae, Gene Expression, Molecular Sequence Data, Protein Conformation.
Teeft :
- Amino acids, Anking sequences, Antigenic, Apc, Bces, Brons, Cassette, Cell epitope, Cell epitopes, Cell line, Cell proliferation assay, Chimeric, Class peptides, Conformation, Different permutational, Epitope, Fournier, General rules, Helical, Helical conformation, Helix, Helix conformation, Hemagglutinin, Hemagglutinin protein, Immune, Immunogenicity, Immunol, Immunological, Immunological methods, Jung, Kasmi, Mabs, Mammalian cells, Maternal antibodies, Measles, Measles virus, Measles virus hemagglutinin protein, Molecular environment, Molecular weight, More antigenic, Multiple copies, Negative control, Neutralizing, Neutralizing antibodies, Peptide, Permutational, Plasmid, Polyepitope, Polyepitopes, Protective antibodies, Recombinant, Rotzschke, Schneider, Semliki forest virus replicon, Sequential, Sequential bces, Sequential epitopes, Stimulation index, Synthetic peptides, Tandem, Tandem tces, Tce, Theisen, Transfected cells, Transmembrane signal sequence, Vaccine, Virol, Western blot, Wiesmuller.

Abstract

Abstract: To investigate a strategy for the design of chimeric antigens based on B cell epitopes (BCEs) we have genetically recombined multiple copies of loop- (L) and helix-forming (H) sequential and protective BCEs of the measles virus hemagglutinin protein (MVH) in a number of high-molecular-weight polyepitope constructs (24.5–45.5 kDa). The BCE cassettes were combined semi-randomly together with a promiscuous T cell epitope (TCE; tt830–844) to yield 13 different permutational constructs. When expressed in mammalian cells, all constructs were detectable by Western blot as distinct bands of predicted molecular weight. Flow cytometry with conformation-specific antibodies revealed the Cys-loop in two [(L4T4)2 and (L2T2)4] and the helix conformation in one [(H2T2)4] of the different permutational constructs. The larger constructs, containing 16 epitope cassettes, seemed more likely to express the BCEs in their native conformation than the 8-mers. In the T cell proliferation assay, constructs with a higher copy number of TCEs, such as (L2T2)4, were more antigenic, as long as tandem repeats were separated by spacers. Since the conformation of even sequential BCEs and the processing of TCEs are both sensitive to their molecular environment it is difficult to predict the antigenic properties of polyepitopes. However, with the permutational approach we have developed several polyepitope constructs [(L4T4)2, (L2T2)4, (H2T2)4] based on complex sequential BCEs that are antigenic for both T and B cells. Several constructs induced sera that reacted with reporter peptides, demonstrating that the sequential nature of the viral epitopes was conserved in the polyepitopes. Although several sera contained antibodies directed against amino acids critical for neutralization, only one construct induced antibodies that cross-reacted with the virus. Our results show the difficulty of designing chimeric antigens based on B cell epitopes mimicking their antigenic and immunologic properties even when these are sequential in nature.

Url:

https://api.istex.fr/ark:/67375/6H6-QQHDZG8L-Z/fulltext.pdf

DOI: 10.1016/S0022-1759(00)00197-6

Affiliations:

Luxembourg (pays), Suisse

Le document en format XML

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<term>Animals</term>
<term>Antigen</term>
<term>Antigenic determinant</term>
<term>Antigenicity</term>
<term>Antigens, Viral (biosynthesis)</term>
<term>Antigens, Viral (chemistry)</term>
<term>Antigens, Viral (genetics)</term>
<term>Antigens, Viral (immunology)</term>
<term>B-Lymphocyte</term>
<term>Cell Line</term>
<term>Chimera</term>
<term>Cricetinae</term>
<term>Epitopes, B-Lymphocyte (biosynthesis)</term>
<term>Epitopes, B-Lymphocyte (chemistry)</term>
<term>Epitopes, B-Lymphocyte (genetics)</term>
<term>Epitopes, B-Lymphocyte (immunology)</term>
<term>Epitopes, T-Lymphocyte (biosynthesis)</term>
<term>Epitopes, T-Lymphocyte (chemistry)</term>
<term>Epitopes, T-Lymphocyte (genetics)</term>
<term>Epitopes, T-Lymphocyte (immunology)</term>
<term>Gene Expression</term>
<term>Helical structure</term>
<term>Hemagglutinin</term>
<term>Hemagglutinins, Viral (biosynthesis)</term>
<term>Hemagglutinins, Viral (chemistry)</term>
<term>Hemagglutinins, Viral (genetics)</term>
<term>Hemagglutinins, Viral (immunology)</term>
<term>Measles virus</term>
<term>Molecular Sequence Data</term>
<term>Peptides (chemistry)</term>
<term>Peptides (genetics)</term>
<term>Peptides (immunology)</term>
<term>Protein</term>
<term>Protein Conformation</term>
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<term>Recombinant Fusion Proteins (genetics)</term>
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<term>Antigènes viraux (biosynthèse)</term>
<term>Antigènes viraux (génétique)</term>
<term>Antigènes viraux (immunologie)</term>
<term>Conformation des protéines</term>
<term>Cricetinae</term>
<term>Données de séquences moléculaires</term>
<term>Déterminants antigéniques des lymphocytes B ()</term>
<term>Déterminants antigéniques des lymphocytes B (biosynthèse)</term>
<term>Déterminants antigéniques des lymphocytes B (génétique)</term>
<term>Déterminants antigéniques des lymphocytes B (immunologie)</term>
<term>Déterminants antigéniques des lymphocytes T ()</term>
<term>Déterminants antigéniques des lymphocytes T (biosynthèse)</term>
<term>Déterminants antigéniques des lymphocytes T (génétique)</term>
<term>Déterminants antigéniques des lymphocytes T (immunologie)</term>
<term>Expression des gènes</term>
<term>Hémagglutinines virales ()</term>
<term>Hémagglutinines virales (biosynthèse)</term>
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<term>Hémagglutinines virales (immunologie)</term>
<term>Lignée cellulaire</term>
<term>Peptides ()</term>
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<term>Protéines de fusion recombinantes (immunologie)</term>
<term>Séquence d'acides aminés</term>
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<term>Hemagglutinins, Viral</term>
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<term>Epitopes, B-Lymphocyte</term>
<term>Epitopes, T-Lymphocyte</term>
<term>Hemagglutinins, Viral</term>
<term>Peptides</term>
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<term>Epitopes, B-Lymphocyte</term>
<term>Epitopes, T-Lymphocyte</term>
<term>Hemagglutinins, Viral</term>
<term>Peptides</term>
<term>Recombinant Fusion Proteins</term>
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<term>Epitopes, B-Lymphocyte</term>
<term>Epitopes, T-Lymphocyte</term>
<term>Hemagglutinins, Viral</term>
<term>Peptides</term>
<term>Recombinant Fusion Proteins</term>
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<term>Déterminants antigéniques des lymphocytes B</term>
<term>Déterminants antigéniques des lymphocytes T</term>
<term>Hémagglutinines virales</term>
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<term>Déterminants antigéniques des lymphocytes B</term>
<term>Déterminants antigéniques des lymphocytes T</term>
<term>Hémagglutinines virales</term>
<term>Peptides</term>
<term>Protéines de fusion recombinantes</term>
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<term>Déterminants antigéniques des lymphocytes B</term>
<term>Déterminants antigéniques des lymphocytes T</term>
<term>Hémagglutinines virales</term>
<term>Peptides</term>
<term>Protéines de fusion recombinantes</term>
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<term>Animals</term>
<term>Cell Line</term>
<term>Cricetinae</term>
<term>Gene Expression</term>
<term>Molecular Sequence Data</term>
<term>Protein Conformation</term>
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<term>Antigène</term>
<term>Antigènes viraux</term>
<term>Antigénicité</term>
<term>Chimère</term>
<term>Conformation des protéines</term>
<term>Cricetinae</term>
<term>Données de séquences moléculaires</term>
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<term>Déterminants antigéniques des lymphocytes B</term>
<term>Déterminants antigéniques des lymphocytes T</term>
<term>Expression des gènes</term>
<term>Hémagglutinine</term>
<term>Hémagglutinines virales</term>
<term>Lignée cellulaire</term>
<term>Lymphocyte B</term>
<term>Lymphocyte T</term>
<term>Peptides</term>
<term>Protéine</term>
<term>Protéines de fusion recombinantes</term>
<term>Structure hélice</term>
<term>Séquence d'acides aminés</term>
<term>Virus rougeole</term>
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<keywords scheme="Teeft" xml:lang="en"><term>Amino acids</term>
<term>Anking sequences</term>
<term>Antigenic</term>
<term>Apc</term>
<term>Bces</term>
<term>Brons</term>
<term>Cassette</term>
<term>Cell epitope</term>
<term>Cell epitopes</term>
<term>Cell line</term>
<term>Cell proliferation assay</term>
<term>Chimeric</term>
<term>Class peptides</term>
<term>Conformation</term>
<term>Different permutational</term>
<term>Epitope</term>
<term>Fournier</term>
<term>General rules</term>
<term>Helical</term>
<term>Helical conformation</term>
<term>Helix</term>
<term>Helix conformation</term>
<term>Hemagglutinin</term>
<term>Hemagglutinin protein</term>
<term>Immune</term>
<term>Immunogenicity</term>
<term>Immunol</term>
<term>Immunological</term>
<term>Immunological methods</term>
<term>Jung</term>
<term>Kasmi</term>
<term>Mabs</term>
<term>Mammalian cells</term>
<term>Maternal antibodies</term>
<term>Measles</term>
<term>Measles virus</term>
<term>Measles virus hemagglutinin protein</term>
<term>Molecular environment</term>
<term>Molecular weight</term>
<term>More antigenic</term>
<term>Multiple copies</term>
<term>Negative control</term>
<term>Neutralizing</term>
<term>Neutralizing antibodies</term>
<term>Peptide</term>
<term>Permutational</term>
<term>Plasmid</term>
<term>Polyepitope</term>
<term>Polyepitopes</term>
<term>Protective antibodies</term>
<term>Recombinant</term>
<term>Rotzschke</term>
<term>Schneider</term>
<term>Semliki forest virus replicon</term>
<term>Sequential</term>
<term>Sequential bces</term>
<term>Sequential epitopes</term>
<term>Stimulation index</term>
<term>Synthetic peptides</term>
<term>Tandem</term>
<term>Tandem tces</term>
<term>Tce</term>
<term>Theisen</term>
<term>Transfected cells</term>
<term>Transmembrane signal sequence</term>
<term>Vaccine</term>
<term>Virol</term>
<term>Western blot</term>
<term>Wiesmuller</term>
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<front><div type="abstract" xml:lang="en">Abstract: To investigate a strategy for the design of chimeric antigens based on B cell epitopes (BCEs) we have genetically recombined multiple copies of loop- (L) and helix-forming (H) sequential and protective BCEs of the measles virus hemagglutinin protein (MVH) in a number of high-molecular-weight polyepitope constructs (24.5–45.5 kDa). The BCE cassettes were combined semi-randomly together with a promiscuous T cell epitope (TCE; tt830–844) to yield 13 different permutational constructs. When expressed in mammalian cells, all constructs were detectable by Western blot as distinct bands of predicted molecular weight. Flow cytometry with conformation-specific antibodies revealed the Cys-loop in two [(L4T4)2 and (L2T2)4] and the helix conformation in one [(H2T2)4] of the different permutational constructs. The larger constructs, containing 16 epitope cassettes, seemed more likely to express the BCEs in their native conformation than the 8-mers. In the T cell proliferation assay, constructs with a higher copy number of TCEs, such as (L2T2)4, were more antigenic, as long as tandem repeats were separated by spacers. Since the conformation of even sequential BCEs and the processing of TCEs are both sensitive to their molecular environment it is difficult to predict the antigenic properties of polyepitopes. However, with the permutational approach we have developed several polyepitope constructs [(L4T4)2, (L2T2)4, (H2T2)4] based on complex sequential BCEs that are antigenic for both T and B cells. Several constructs induced sera that reacted with reporter peptides, demonstrating that the sequential nature of the viral epitopes was conserved in the polyepitopes. Although several sera contained antibodies directed against amino acids critical for neutralization, only one construct induced antibodies that cross-reacted with the virus. Our results show the difficulty of designing chimeric antigens based on B cell epitopes mimicking their antigenic and immunologic properties even when these are sequential in nature.</div>
</front>
</TEI>
<affiliations><list><country><li>Luxembourg (pays)</li>
<li>Suisse</li>
</country>
</list>
<tree><country name="Luxembourg (pays)"><noRegion><name sortKey="Theisen, D M" sort="Theisen, D M" uniqKey="Theisen D" first="D. M" last="Theisen">D. M Theisen</name>
</noRegion>
<name sortKey="Ammerlaan, W" sort="Ammerlaan, W" uniqKey="Ammerlaan W" first="W" last="Ammerlaan">W. Ammerlaan</name>
<name sortKey="Bouche, F B" sort="Bouche, F B" uniqKey="Bouche F" first="F. B" last="Bouche">F. B Bouche</name>
<name sortKey="El Kasmi, K C" sort="El Kasmi, K C" uniqKey="El Kasmi K" first="K. C" last="El Kasmi">K. C El Kasmi</name>
<name sortKey="Muller, C P" sort="Muller, C P" uniqKey="Muller C" first="C. P" last="Muller">C. P Muller</name>
<name sortKey="Muller, C P" sort="Muller, C P" uniqKey="Muller C" first="C. P" last="Muller">C. P Muller</name>
<name sortKey="Von Der Ahe, I" sort="Von Der Ahe, I" uniqKey="Von Der Ahe I" first="I" last="Von Der Ahe">I. Von Der Ahe</name>
</country>
<country name="Suisse"><noRegion><name sortKey="Demotz, S" sort="Demotz, S" uniqKey="Demotz S" first="S" last="Demotz">S. Demotz</name>
</noRegion>
</country>
</tree>
</affiliations>
</record>

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Differential antigenicity of recombinant polyepitope-antigens based on loop- and helix-forming B and T cell epitopes

Differential antigenicity of recombinant polyepitope-antigens based on loop- and helix-forming B and T cell epitopes

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