Differential antigenicity of recombinant polyepitope-antigens based on loop- and helix-forming B and T cell epitopes
Identifieur interne : 000094 ( PascalFrancis/Checkpoint ); précédent : 000093; suivant : 000095Differential antigenicity of recombinant polyepitope-antigens based on loop- and helix-forming B and T cell epitopes
Auteurs : D. M. Theisen [Luxembourg (pays), Allemagne] ; F. B. Bouche [Luxembourg (pays), Belgique] ; K. C. El Kasmi [Luxembourg (pays), Allemagne] ; I. Von Der Ahe [Luxembourg (pays), Allemagne] ; W. Ammerlaan [Luxembourg (pays)] ; S. Demotz [Suisse] ; C. P. Muller [Luxembourg (pays), Allemagne]Source :
- Journal of immunological methods [ 0022-1759 ] ; 2000.
Descripteurs français
- Pascal (Inist)
English descriptors
- KwdEn :
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.
Affiliations:
- Allemagne, Belgique, Luxembourg (pays), Suisse
- Bade-Wurtemberg, District de Tübingen, Région de Bruxelles-Capitale
- Bruxelles, Tübingen
- Université libre de Bruxelles
Links toward previous steps (curation, corpus...)
Links to Exploration step
Pascal:01-0262336Le document en format XML
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en" level="a">Differential antigenicity of recombinant polyepitope-antigens based on loop- and helix-forming B and T cell epitopes</title><author><name sortKey="Theisen, D M" sort="Theisen, D M" uniqKey="Theisen D" first="D. M." last="Theisen">D. M. Theisen</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Department of Immunology and WHO Collaborating Center for Measles, Laboratoire National de Santé, B.P. 1102</s1><s2>1011 Luxembourg</s2><s3>LUX</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>Luxembourg (pays)</country><wicri:noRegion>1011 Luxembourg</wicri:noRegion></affiliation><affiliation wicri:level="3"><inist:fA14 i1="02"><s1>Fakultät für Biologie, University of Tübingen</s1><s2>Tübingen</s2><s3>DEU</s3><sZ>1 aut.</sZ></inist:fA14><country>Allemagne</country><placeName><region type="land" nuts="1">Bade-Wurtemberg</region><region type="district" nuts="2">District de Tübingen</region><settlement type="city">Tübingen</settlement></placeName></affiliation></author><author><name sortKey="Bouche, F B" sort="Bouche, F B" uniqKey="Bouche F" first="F. B." last="Bouche">F. B. Bouche</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Department of Immunology and WHO Collaborating Center for Measles, Laboratoire National de Santé, B.P. 1102</s1><s2>1011 Luxembourg</s2><s3>LUX</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>Luxembourg (pays)</country><wicri:noRegion>1011 Luxembourg</wicri:noRegion></affiliation><affiliation wicri:level="4"><inist:fA14 i1="03"><s1>Service de Génétique Appliquée, Université Libre de Bruxelles</s1><s2>Nivelles</s2><s3>BEL</s3><sZ>2 aut.</sZ></inist:fA14><country>Belgique</country><placeName><settlement type="city">Bruxelles</settlement><region type="region" nuts="2">Région de Bruxelles-Capitale</region></placeName><orgName type="university">Université libre de Bruxelles</orgName></affiliation></author><author><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><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Department of Immunology and WHO Collaborating Center for Measles, Laboratoire National de Santé, B.P. 1102</s1><s2>1011 Luxembourg</s2><s3>LUX</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>Luxembourg (pays)</country><wicri:noRegion>1011 Luxembourg</wicri:noRegion></affiliation><affiliation wicri:level="3"><inist:fA14 i1="04"><s1>Medizinische Fakultät, University of Tübingen</s1><s2>Tübingen</s2><s3>DEU</s3><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>Allemagne</country><placeName><region type="land" nuts="1">Bade-Wurtemberg</region><region type="district" nuts="2">District de Tübingen</region><settlement type="city">Tübingen</settlement></placeName></affiliation></author><author><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><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Department of Immunology and WHO Collaborating Center for Measles, Laboratoire National de Santé, B.P. 1102</s1><s2>1011 Luxembourg</s2><s3>LUX</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>Luxembourg (pays)</country><wicri:noRegion>1011 Luxembourg</wicri:noRegion></affiliation><affiliation wicri:level="3"><inist:fA14 i1="04"><s1>Medizinische Fakultät, University of Tübingen</s1><s2>Tübingen</s2><s3>DEU</s3><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>Allemagne</country><placeName><region type="land" nuts="1">Bade-Wurtemberg</region><region type="district" nuts="2">District de Tübingen</region><settlement type="city">Tübingen</settlement></placeName></affiliation></author><author><name sortKey="Ammerlaan, W" sort="Ammerlaan, W" uniqKey="Ammerlaan W" first="W." last="Ammerlaan">W. Ammerlaan</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Department of Immunology and WHO Collaborating Center for Measles, Laboratoire National de Santé, B.P. 1102</s1><s2>1011 Luxembourg</s2><s3>LUX</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>Luxembourg (pays)</country><wicri:noRegion>1011 Luxembourg</wicri:noRegion></affiliation></author><author><name sortKey="Demotz, S" sort="Demotz, S" uniqKey="Demotz S" first="S." last="Demotz">S. Demotz</name><affiliation wicri:level="1"><inist:fA14 i1="05"><s1>Institut de Biochemie, University of Lausanne</s1><s2>Epalinges</s2><s3>CHE</s3><sZ>6 aut.</sZ></inist:fA14><country>Suisse</country><wicri:noRegion>Epalinges</wicri:noRegion></affiliation></author><author><name sortKey="Muller, C P" sort="Muller, C P" uniqKey="Muller C" first="C. P." last="Muller">C. P. Muller</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Department of Immunology and WHO Collaborating Center for Measles, Laboratoire National de Santé, B.P. 1102</s1><s2>1011 Luxembourg</s2><s3>LUX</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>Luxembourg (pays)</country><wicri:noRegion>1011 Luxembourg</wicri:noRegion></affiliation><affiliation wicri:level="3"><inist:fA14 i1="04"><s1>Medizinische Fakultät, University of Tübingen</s1><s2>Tübingen</s2><s3>DEU</s3><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>Allemagne</country><placeName><region type="land" nuts="1">Bade-Wurtemberg</region><region type="district" nuts="2">District de Tübingen</region><settlement type="city">Tübingen</settlement></placeName></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">INIST</idno><idno type="inist">01-0262336</idno><date when="2000">2000</date><idno type="stanalyst">PASCAL 01-0262336 INIST</idno><idno type="RBID">Pascal:01-0262336</idno><idno type="wicri:Area/PascalFrancis/Corpus">000091</idno><idno type="wicri:Area/PascalFrancis/Curation">000003</idno><idno type="wicri:Area/PascalFrancis/Checkpoint">000094</idno><idno type="wicri:explorRef" wicri:stream="PascalFrancis" wicri:step="Checkpoint">000094</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a">Differential antigenicity of recombinant polyepitope-antigens based on loop- and helix-forming B and T cell epitopes</title><author><name sortKey="Theisen, D M" sort="Theisen, D M" uniqKey="Theisen D" first="D. M." last="Theisen">D. M. Theisen</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Department of Immunology and WHO Collaborating Center for Measles, Laboratoire National de Santé, B.P. 1102</s1><s2>1011 Luxembourg</s2><s3>LUX</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>Luxembourg (pays)</country><wicri:noRegion>1011 Luxembourg</wicri:noRegion></affiliation><affiliation wicri:level="3"><inist:fA14 i1="02"><s1>Fakultät für Biologie, University of Tübingen</s1><s2>Tübingen</s2><s3>DEU</s3><sZ>1 aut.</sZ></inist:fA14><country>Allemagne</country><placeName><region type="land" nuts="1">Bade-Wurtemberg</region><region type="district" nuts="2">District de Tübingen</region><settlement type="city">Tübingen</settlement></placeName></affiliation></author><author><name sortKey="Bouche, F B" sort="Bouche, F B" uniqKey="Bouche F" first="F. B." last="Bouche">F. B. Bouche</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Department of Immunology and WHO Collaborating Center for Measles, Laboratoire National de Santé, B.P. 1102</s1><s2>1011 Luxembourg</s2><s3>LUX</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>Luxembourg (pays)</country><wicri:noRegion>1011 Luxembourg</wicri:noRegion></affiliation><affiliation wicri:level="4"><inist:fA14 i1="03"><s1>Service de Génétique Appliquée, Université Libre de Bruxelles</s1><s2>Nivelles</s2><s3>BEL</s3><sZ>2 aut.</sZ></inist:fA14><country>Belgique</country><placeName><settlement type="city">Bruxelles</settlement><region type="region" nuts="2">Région de Bruxelles-Capitale</region></placeName><orgName type="university">Université libre de Bruxelles</orgName></affiliation></author><author><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><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Department of Immunology and WHO Collaborating Center for Measles, Laboratoire National de Santé, B.P. 1102</s1><s2>1011 Luxembourg</s2><s3>LUX</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>Luxembourg (pays)</country><wicri:noRegion>1011 Luxembourg</wicri:noRegion></affiliation><affiliation wicri:level="3"><inist:fA14 i1="04"><s1>Medizinische Fakultät, University of Tübingen</s1><s2>Tübingen</s2><s3>DEU</s3><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>Allemagne</country><placeName><region type="land" nuts="1">Bade-Wurtemberg</region><region type="district" nuts="2">District de Tübingen</region><settlement type="city">Tübingen</settlement></placeName></affiliation></author><author><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><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Department of Immunology and WHO Collaborating Center for Measles, Laboratoire National de Santé, B.P. 1102</s1><s2>1011 Luxembourg</s2><s3>LUX</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>Luxembourg (pays)</country><wicri:noRegion>1011 Luxembourg</wicri:noRegion></affiliation><affiliation wicri:level="3"><inist:fA14 i1="04"><s1>Medizinische Fakultät, University of Tübingen</s1><s2>Tübingen</s2><s3>DEU</s3><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>Allemagne</country><placeName><region type="land" nuts="1">Bade-Wurtemberg</region><region type="district" nuts="2">District de Tübingen</region><settlement type="city">Tübingen</settlement></placeName></affiliation></author><author><name sortKey="Ammerlaan, W" sort="Ammerlaan, W" uniqKey="Ammerlaan W" first="W." last="Ammerlaan">W. Ammerlaan</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Department of Immunology and WHO Collaborating Center for Measles, Laboratoire National de Santé, B.P. 1102</s1><s2>1011 Luxembourg</s2><s3>LUX</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>Luxembourg (pays)</country><wicri:noRegion>1011 Luxembourg</wicri:noRegion></affiliation></author><author><name sortKey="Demotz, S" sort="Demotz, S" uniqKey="Demotz S" first="S." last="Demotz">S. Demotz</name><affiliation wicri:level="1"><inist:fA14 i1="05"><s1>Institut de Biochemie, University of Lausanne</s1><s2>Epalinges</s2><s3>CHE</s3><sZ>6 aut.</sZ></inist:fA14><country>Suisse</country><wicri:noRegion>Epalinges</wicri:noRegion></affiliation></author><author><name sortKey="Muller, C P" sort="Muller, C P" uniqKey="Muller C" first="C. P." last="Muller">C. P. Muller</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Department of Immunology and WHO Collaborating Center for Measles, Laboratoire National de Santé, B.P. 1102</s1><s2>1011 Luxembourg</s2><s3>LUX</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>Luxembourg (pays)</country><wicri:noRegion>1011 Luxembourg</wicri:noRegion></affiliation><affiliation wicri:level="3"><inist:fA14 i1="04"><s1>Medizinische Fakultät, University of Tübingen</s1><s2>Tübingen</s2><s3>DEU</s3><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>Allemagne</country><placeName><region type="land" nuts="1">Bade-Wurtemberg</region><region type="district" nuts="2">District de Tübingen</region><settlement type="city">Tübingen</settlement></placeName></affiliation></author></analytic><series><title level="j" type="main">Journal of immunological methods</title><title level="j" type="abbreviated">J. immunol. methods</title><idno type="ISSN">0022-1759</idno><imprint><date when="2000">2000</date></imprint></series></biblStruct></sourceDesc><seriesStmt><title level="j" type="main">Journal of immunological methods</title><title level="j" type="abbreviated">J. immunol. methods</title><idno type="ISSN">0022-1759</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Antigen</term><term>Antigenic determinant</term><term>Antigenicity</term><term>B-Lymphocyte</term><term>Chimera</term><term>Helical structure</term><term>Hemagglutinin</term><term>Measles virus</term><term>Protein</term><term>T-Lymphocyte</term></keywords><keywords scheme="Pascal" xml:lang="fr"><term>Lymphocyte T</term><term>Lymphocyte B</term><term>Déterminant antigénique</term><term>Virus rougeole</term><term>Antigène</term><term>Chimère</term><term>Protéine</term><term>Hémagglutinine</term><term>Structure hélice</term><term>Antigénicité</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">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 [(L<sub>4</sub>T<sub>4</sub>)<sub>2</sub> and (L<sub>2</sub>T<sub>2</sub>)<sub>4</sub>] and the helix conformation in one [(H<sub>2</sub>T<sub>2</sub>)<sub>4</sub>] 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 (L<sub>2</sub>T<sub>2</sub>)<sub>4</sub> 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 [(L<sub>4</sub>T<sub>4</sub>)<sub>2</sub>, (L<sub>2</sub>T<sub>2</sub>)<sub>4</sub>, (H<sub>2</sub>T<sub>2</sub>)<sub>4</sub>] 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><inist><standard h6="B"><pA><fA01 i1="01" i2="1"><s0>0022-1759</s0></fA01><fA02 i1="01"><s0>JIMMBG</s0></fA02><fA03 i2="1"><s0>J. immunol. methods</s0></fA03><fA05><s2>242</s2></fA05><fA06><s2>1-2</s2></fA06><fA08 i1="01" i2="1" l="ENG"><s1>Differential antigenicity of recombinant polyepitope-antigens based on loop- and helix-forming B and T cell epitopes</s1></fA08><fA11 i1="01" i2="1"><s1>THEISEN (D. M.)</s1></fA11><fA11 i1="02" i2="1"><s1>BOUCHE (F. B.)</s1></fA11><fA11 i1="03" i2="1"><s1>EL KASMI (K. C.)</s1></fA11><fA11 i1="04" i2="1"><s1>VON DER AHE (I.)</s1></fA11><fA11 i1="05" i2="1"><s1>AMMERLAAN (W.)</s1></fA11><fA11 i1="06" i2="1"><s1>DEMOTZ (S.)</s1></fA11><fA11 i1="07" i2="1"><s1>MULLER (C. P.)</s1></fA11><fA14 i1="01"><s1>Department of Immunology and WHO Collaborating Center for Measles, Laboratoire National de Santé, B.P. 1102</s1><s2>1011 Luxembourg</s2><s3>LUX</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>7 aut.</sZ></fA14><fA14 i1="02"><s1>Fakultät für Biologie, University of Tübingen</s1><s2>Tübingen</s2><s3>DEU</s3><sZ>1 aut.</sZ></fA14><fA14 i1="03"><s1>Service de Génétique Appliquée, Université Libre de Bruxelles</s1><s2>Nivelles</s2><s3>BEL</s3><sZ>2 aut.</sZ></fA14><fA14 i1="04"><s1>Medizinische Fakultät, University of Tübingen</s1><s2>Tübingen</s2><s3>DEU</s3><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>7 aut.</sZ></fA14><fA14 i1="05"><s1>Institut de Biochemie, University of Lausanne</s1><s2>Epalinges</s2><s3>CHE</s3><sZ>6 aut.</sZ></fA14><fA20><s1>145-157</s1></fA20><fA21><s1>2000</s1></fA21><fA23 i1="01"><s0>ENG</s0></fA23><fA43 i1="01"><s1>INIST</s1><s2>15654</s2><s5>354000090666720140</s5></fA43><fA44><s0>0000</s0><s1>© 2001 INIST-CNRS. All rights reserved.</s1></fA44><fA45><s0>1 p.1/2</s0></fA45><fA47 i1="01" i2="1"><s0>01-0262336</s0></fA47><fA60><s1>P</s1></fA60><fA61><s0>A</s0></fA61><fA64 i1="01" i2="1"><s0>Journal of immunological methods</s0></fA64><fA66 i1="01"><s0>NLD</s0></fA66><fC01 i1="01" l="ENG"><s0>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 [(L<sub>4</sub>T<sub>4</sub>)<sub>2</sub> and (L<sub>2</sub>T<sub>2</sub>)<sub>4</sub>] and the helix conformation in one [(H<sub>2</sub>T<sub>2</sub>)<sub>4</sub>] 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 (L<sub>2</sub>T<sub>2</sub>)<sub>4</sub> 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 [(L<sub>4</sub>T<sub>4</sub>)<sub>2</sub>, (L<sub>2</sub>T<sub>2</sub>)<sub>4</sub>, (H<sub>2</sub>T<sub>2</sub>)<sub>4</sub>] 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.</s0></fC01><fC02 i1="01" i2="X"><s0>002A05C10</s0></fC02><fC03 i1="01" i2="X" l="FRE"><s0>Lymphocyte T</s0><s5>01</s5></fC03><fC03 i1="01" i2="X" l="ENG"><s0>T-Lymphocyte</s0><s5>01</s5><s6>«T»-Lymphocyte</s6></fC03><fC03 i1="01" i2="X" l="SPA"><s0>Linfocito T</s0><s5>01</s5></fC03><fC03 i1="02" i2="X" l="FRE"><s0>Lymphocyte B</s0><s5>02</s5></fC03><fC03 i1="02" i2="X" l="ENG"><s0>B-Lymphocyte</s0><s5>02</s5><s6>«B»-Lymphocyte</s6></fC03><fC03 i1="02" i2="X" l="SPA"><s0>Linfocito B</s0><s5>02</s5></fC03><fC03 i1="03" i2="X" l="FRE"><s0>Déterminant antigénique</s0><s5>03</s5></fC03><fC03 i1="03" i2="X" l="ENG"><s0>Antigenic determinant</s0><s5>03</s5></fC03><fC03 i1="03" i2="X" l="SPA"><s0>Determinante antigénico</s0><s5>03</s5></fC03><fC03 i1="04" i2="X" l="FRE"><s0>Virus rougeole</s0><s2>NW</s2><s5>04</s5></fC03><fC03 i1="04" i2="X" l="ENG"><s0>Measles virus</s0><s2>NW</s2><s5>04</s5></fC03><fC03 i1="04" i2="X" l="SPA"><s0>Measles virus</s0><s2>NW</s2><s5>04</s5></fC03><fC03 i1="05" i2="X" l="FRE"><s0>Antigène</s0><s5>05</s5></fC03><fC03 i1="05" i2="X" l="ENG"><s0>Antigen</s0><s5>05</s5></fC03><fC03 i1="05" i2="X" l="SPA"><s0>Antígeno</s0><s5>05</s5></fC03><fC03 i1="06" i2="X" l="FRE"><s0>Chimère</s0><s5>06</s5></fC03><fC03 i1="06" i2="X" l="ENG"><s0>Chimera</s0><s5>06</s5></fC03><fC03 i1="06" i2="X" l="SPA"><s0>Quimera</s0><s5>06</s5></fC03><fC03 i1="07" i2="X" l="FRE"><s0>Protéine</s0><s5>07</s5></fC03><fC03 i1="07" i2="X" l="ENG"><s0>Protein</s0><s5>07</s5></fC03><fC03 i1="07" i2="X" l="SPA"><s0>Proteína</s0><s5>07</s5></fC03><fC03 i1="08" i2="X" l="FRE"><s0>Hémagglutinine</s0><s5>08</s5></fC03><fC03 i1="08" i2="X" l="ENG"><s0>Hemagglutinin</s0><s5>08</s5></fC03><fC03 i1="08" i2="X" l="SPA"><s0>Hemoaglutinina</s0><s5>08</s5></fC03><fC03 i1="09" i2="X" l="FRE"><s0>Structure hélice</s0><s5>09</s5></fC03><fC03 i1="09" i2="X" l="ENG"><s0>Helical structure</s0><s5>09</s5></fC03><fC03 i1="09" i2="X" l="SPA"><s0>Estructura helicoidal</s0><s5>09</s5></fC03><fC03 i1="10" i2="X" l="FRE"><s0>Antigénicité</s0><s5>10</s5></fC03><fC03 i1="10" i2="X" l="ENG"><s0>Antigenicity</s0><s5>10</s5></fC03><fC03 i1="10" i2="X" l="SPA"><s0>Antigenicidad</s0><s5>10</s5></fC03><fC07 i1="01" i2="X" l="FRE"><s0>Morbillivirus</s0><s2>NW</s2></fC07><fC07 i1="01" i2="X" l="ENG"><s0>Morbillivirus</s0><s2>NW</s2></fC07><fC07 i1="01" i2="X" l="SPA"><s0>Morbillivirus</s0><s2>NW</s2></fC07><fC07 i1="02" i2="X" l="FRE"><s0>Paramyxovirinae</s0><s2>NW</s2></fC07><fC07 i1="02" i2="X" l="ENG"><s0>Paramyxovirinae</s0><s2>NW</s2></fC07><fC07 i1="02" i2="X" l="SPA"><s0>Paramyxovirinae</s0><s2>NW</s2></fC07><fC07 i1="03" i2="X" l="FRE"><s0>Paramyxoviridae</s0><s2>NW</s2></fC07><fC07 i1="03" i2="X" l="ENG"><s0>Paramyxoviridae</s0><s2>NW</s2></fC07><fC07 i1="03" i2="X" l="SPA"><s0>Paramyxoviridae</s0><s2>NW</s2></fC07><fC07 i1="04" i2="X" l="FRE"><s0>Mononegavirales</s0><s2>NW</s2></fC07><fC07 i1="04" i2="X" l="ENG"><s0>Mononegavirales</s0><s2>NW</s2></fC07><fC07 i1="04" i2="X" l="SPA"><s0>Mononegavirales</s0><s2>NW</s2></fC07><fC07 i1="05" i2="X" l="FRE"><s0>Virus</s0><s2>NW</s2></fC07><fC07 i1="05" i2="X" l="ENG"><s0>Virus</s0><s2>NW</s2></fC07><fC07 i1="05" i2="X" l="SPA"><s0>Virus</s0><s2>NW</s2></fC07><fC07 i1="06" i2="X" l="FRE"><s0>Glycoprotéine</s0><s5>28</s5></fC07><fC07 i1="06" i2="X" l="ENG"><s0>Glycoprotein</s0><s5>28</s5></fC07><fC07 i1="06" i2="X" l="SPA"><s0>Glicoproteína</s0><s5>28</s5></fC07><fN21><s1>183</s1></fN21></pA></standard></inist><affiliations><list><country><li>Allemagne</li><li>Belgique</li><li>Luxembourg (pays)</li><li>Suisse</li></country><region><li>Bade-Wurtemberg</li><li>District de Tübingen</li><li>Région de Bruxelles-Capitale</li></region><settlement><li>Bruxelles</li><li>Tübingen</li></settlement><orgName><li>Université libre de Bruxelles</li></orgName></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="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="Allemagne"><region name="Bade-Wurtemberg"><name sortKey="Theisen, D M" sort="Theisen, D M" uniqKey="Theisen D" first="D. M." last="Theisen">D. M. Theisen</name></region><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="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="Belgique"><region name="Région de Bruxelles-Capitale"><name sortKey="Bouche, F B" sort="Bouche, F B" uniqKey="Bouche F" first="F. B." last="Bouche">F. B. Bouche</name></region></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>Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=$WICRI_ROOT/Sante/explor/MersV1/Data/PascalFrancis/Checkpoint
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 000094 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/PascalFrancis/Checkpoint/biblio.hfd -nk 000094 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien
|wiki= Sante
|area= MersV1
|flux= PascalFrancis
|étape= Checkpoint
|type= RBID
|clé= Pascal:01-0262336
|texte= Differential antigenicity of recombinant polyepitope-antigens based on loop- and helix-forming B and T cell epitopes
}}
| This area was generated with Dilib version V0.6.33. |  |