Development and characterization of neutralizing monoclonal antibodies against the pandemic H1N1 virus (2009).
Identifieur interne : 001B54 ( Main/Exploration ); précédent : 001B53; suivant : 001B55Development and characterization of neutralizing monoclonal antibodies against the pandemic H1N1 virus (2009).
Auteurs : Teresa M. Cabral [Canada] ; Yohannes Berhane ; Lisa Schmidt ; Dobryan M. Tracz ; Kate Hole ; Marsha Leith ; Cindi R. CorbettSource :
- Journal of virological methods [ 1879-0984 ] ; 2012.
Descripteurs français
- KwdFr :
- Affinité des anticorps, Animaux, Anticorps antiviraux (immunologie), Anticorps antiviraux (isolement et purification), Anticorps antiviraux (usage thérapeutique), Anticorps monoclonaux (immunologie), Anticorps monoclonaux (isolement et purification), Anticorps monoclonaux (usage thérapeutique), Anticorps neutralisants (immunologie), Anticorps neutralisants (isolement et purification), Anticorps neutralisants (usage thérapeutique), Femelle, Glycoprotéine hémagglutinine du virus influenza (immunologie), Humains, Immunoglobuline G (immunologie), Immunoglobuline G (isolement et purification), Immunoglobuline G (usage thérapeutique), Immunothérapie (), Réactions croisées, Sensibilité et spécificité, Souris, Souris de lignée BALB C, Sous-type H1N1 du virus de la grippe A (), Sous-type H1N1 du virus de la grippe A (immunologie), Sous-type H1N1 du virus de la grippe A (isolement et purification), Test ELISA ().
- MESH :
- immunologie : Anticorps antiviraux, Anticorps monoclonaux, Anticorps neutralisants, Glycoprotéine hémagglutinine du virus influenza, Immunoglobuline G, Sous-type H1N1 du virus de la grippe A.
- isolement et purification : Anticorps antiviraux, Anticorps monoclonaux, Anticorps neutralisants, Immunoglobuline G, Sous-type H1N1 du virus de la grippe A.
- usage thérapeutique : Anticorps antiviraux, Anticorps monoclonaux, Anticorps neutralisants, Immunoglobuline G.
- Affinité des anticorps, Animaux, Femelle, Humains, Immunothérapie, Réactions croisées, Sensibilité et spécificité, Souris, Souris de lignée BALB C, Sous-type H1N1 du virus de la grippe A, Test ELISA.
English descriptors
- KwdEn :
- Animals, Antibodies, Monoclonal (immunology), Antibodies, Monoclonal (isolation & purification), Antibodies, Monoclonal (therapeutic use), Antibodies, Neutralizing (immunology), Antibodies, Neutralizing (isolation & purification), Antibodies, Neutralizing (therapeutic use), Antibodies, Viral (immunology), Antibodies, Viral (isolation & purification), Antibodies, Viral (therapeutic use), Antibody Affinity, Cross Reactions, Enzyme-Linked Immunosorbent Assay (methods), Female, Hemagglutinin Glycoproteins, Influenza Virus (immunology), Humans, Immunoglobulin G (immunology), Immunoglobulin G (isolation & purification), Immunoglobulin G (therapeutic use), Immunotherapy (methods), Influenza A Virus, H1N1 Subtype (drug effects), Influenza A Virus, H1N1 Subtype (immunology), Influenza A Virus, H1N1 Subtype (isolation & purification), Mice, Mice, Inbred BALB C, Sensitivity and Specificity.
- MESH :
- chemical , immunology : Antibodies, Monoclonal, Antibodies, Neutralizing, Antibodies, Viral, Hemagglutinin Glycoproteins, Influenza Virus, Immunoglobulin G.
- chemical , isolation & purification : Antibodies, Monoclonal, Antibodies, Neutralizing, Antibodies, Viral, Immunoglobulin G.
- chemical , therapeutic use : Antibodies, Monoclonal, Antibodies, Neutralizing, Antibodies, Viral, Immunoglobulin G.
- drug effects : Influenza A Virus, H1N1 Subtype.
- immunology : Influenza A Virus, H1N1 Subtype.
- isolation & purification : Influenza A Virus, H1N1 Subtype.
- methods : Enzyme-Linked Immunosorbent Assay, Immunotherapy.
- Animals, Antibody Affinity, Cross Reactions, Female, Humans, Mice, Mice, Inbred BALB C, Sensitivity and Specificity.
Abstract
The 2009 H1N1 influenza pandemic was a major international public health crisis which caused considerable morbidity and mortality worldwide. The goal of this study was to produce anti-H1 monoclonal antibodies (MAbs) for improving diagnostic immunological assays and to develop potential immunotherapeutics. Nine MAbs were produced after immunizing mice with recombinant hemagglutinin (HA) protein from A/California/06/09. Two spleenocyte myeloma fusions yielded 1588 hybridoma cultures. After screening the hybridoma culture supernatants for antibody reactivity to rHA, nine clones were selected for further characterization. Cross-reactivity studies of the anti-rHA antibodies against a panel of influenza viruses (H1-H16) revealed eight out of nine MAbs were specific to the pandemic H1 subtype, except for MAb F256G2sc1 which also cross-reacted with H5 subtype virus. All MAbs were of the IgG1κ isotype, except F256G2sc1 which was IgG2aκ. The anti-rHA MAbs had binding affinities to rHA that ranged from a K(D) (disassociation constant) of 1.34×10(-9)M (F255G7sc1) to the weakest affinity of 4.60×10(-8)M (F255G4sc1). Interestingly, in a plaque reduction neutralization assay, all MAbs except F255G3sc1 demonstrated neutralizing ability. Furthermore, all MAbs except F255G3sc1 and F255G9sc1 exhibited anti-hemagglutinin activity against pandemic H1N1 viruses, but not against classical North American swine influenza viruses of the same subtype. Immunofluorescence assay (IFA) demonstrated that all MAbs except F255G1sc1 and F255G3sc1 were able to detect 2009 pandemic H1N1 (2009) virus- infected MDCK cells. The MAbs were also evaluated for potential use in competitive ELISA (cELISA), and with the exception of F255G3sc1, all MAbs showed competitive activity with serum collected from pigs infected with pandemic H1N1 virus (2009). The developed MAbs have demonstrated utility as immunodiagnostic and research reagents, and their neutralizing capabilities also hold potential for designing antiviral drugs against pandemic influenza.
DOI: 10.1016/j.jviromet.2012.03.016
PubMed: 22575685
Affiliations:
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Cabral</name><affiliation wicri:level="1"><nlm:affiliation>National Microbiology Laboratory, Public Health Agency of Canada, 1015 Arlington Street, Winnipeg, Manitoba, R3E 3R2, Canada.</nlm:affiliation><country xml:lang="fr">Canada</country><wicri:regionArea>National Microbiology Laboratory, Public Health Agency of Canada, 1015 Arlington Street, Winnipeg, Manitoba, R3E 3R2</wicri:regionArea><wicri:noRegion>R3E 3R2</wicri:noRegion></affiliation></author><author><name sortKey="Berhane, Yohannes" sort="Berhane, Yohannes" uniqKey="Berhane Y" first="Yohannes" last="Berhane">Yohannes Berhane</name></author><author><name sortKey="Schmidt, Lisa" sort="Schmidt, Lisa" uniqKey="Schmidt L" first="Lisa" last="Schmidt">Lisa Schmidt</name></author><author><name sortKey="Tracz, Dobryan M" sort="Tracz, Dobryan M" uniqKey="Tracz D" first="Dobryan M" last="Tracz">Dobryan M. Tracz</name></author><author><name sortKey="Hole, Kate" sort="Hole, Kate" uniqKey="Hole K" first="Kate" last="Hole">Kate Hole</name></author><author><name sortKey="Leith, Marsha" sort="Leith, Marsha" uniqKey="Leith M" first="Marsha" last="Leith">Marsha Leith</name></author><author><name sortKey="Corbett, Cindi R" sort="Corbett, Cindi R" uniqKey="Corbett C" first="Cindi R" last="Corbett">Cindi R. 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Tracz</name></author><author><name sortKey="Hole, Kate" sort="Hole, Kate" uniqKey="Hole K" first="Kate" last="Hole">Kate Hole</name></author><author><name sortKey="Leith, Marsha" sort="Leith, Marsha" uniqKey="Leith M" first="Marsha" last="Leith">Marsha Leith</name></author><author><name sortKey="Corbett, Cindi R" sort="Corbett, Cindi R" uniqKey="Corbett C" first="Cindi R" last="Corbett">Cindi R. Corbett</name></author></analytic><series><title level="j">Journal of virological methods</title><idno type="eISSN">1879-0984</idno><imprint><date when="2012" type="published">2012</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term><term>Antibodies, Monoclonal (immunology)</term><term>Antibodies, Monoclonal (isolation & purification)</term><term>Antibodies, Monoclonal (therapeutic use)</term><term>Antibodies, Neutralizing (immunology)</term><term>Antibodies, Neutralizing (isolation & purification)</term><term>Antibodies, Neutralizing (therapeutic use)</term><term>Antibodies, Viral (immunology)</term><term>Antibodies, Viral (isolation & purification)</term><term>Antibodies, Viral (therapeutic use)</term><term>Antibody Affinity</term><term>Cross Reactions</term><term>Enzyme-Linked Immunosorbent Assay (methods)</term><term>Female</term><term>Hemagglutinin Glycoproteins, Influenza Virus (immunology)</term><term>Humans</term><term>Immunoglobulin G (immunology)</term><term>Immunoglobulin G (isolation & purification)</term><term>Immunoglobulin G (therapeutic use)</term><term>Immunotherapy (methods)</term><term>Influenza A Virus, H1N1 Subtype (drug effects)</term><term>Influenza A Virus, H1N1 Subtype (immunology)</term><term>Influenza A Virus, H1N1 Subtype (isolation & purification)</term><term>Mice</term><term>Mice, Inbred BALB C</term><term>Sensitivity and Specificity</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Affinité des anticorps</term><term>Animaux</term><term>Anticorps antiviraux (immunologie)</term><term>Anticorps antiviraux (isolement et purification)</term><term>Anticorps antiviraux (usage thérapeutique)</term><term>Anticorps monoclonaux (immunologie)</term><term>Anticorps monoclonaux (isolement et purification)</term><term>Anticorps monoclonaux (usage thérapeutique)</term><term>Anticorps neutralisants (immunologie)</term><term>Anticorps neutralisants (isolement et purification)</term><term>Anticorps neutralisants (usage thérapeutique)</term><term>Femelle</term><term>Glycoprotéine hémagglutinine du virus influenza (immunologie)</term><term>Humains</term><term>Immunoglobuline G (immunologie)</term><term>Immunoglobuline G (isolement et purification)</term><term>Immunoglobuline G (usage thérapeutique)</term><term>Immunothérapie ()</term><term>Réactions croisées</term><term>Sensibilité et spécificité</term><term>Souris</term><term>Souris de lignée BALB C</term><term>Sous-type H1N1 du virus de la grippe A ()</term><term>Sous-type H1N1 du virus de la grippe A (immunologie)</term><term>Sous-type H1N1 du virus de la grippe A (isolement et purification)</term><term>Test ELISA ()</term></keywords><keywords scheme="MESH" type="chemical" qualifier="immunology" xml:lang="en"><term>Antibodies, Monoclonal</term><term>Antibodies, Neutralizing</term><term>Antibodies, Viral</term><term>Hemagglutinin Glycoproteins, Influenza 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xml:lang="en"><term>Influenza A Virus, H1N1 Subtype</term></keywords><keywords scheme="MESH" qualifier="isolation & purification" xml:lang="en"><term>Influenza A Virus, H1N1 Subtype</term></keywords><keywords scheme="MESH" qualifier="isolement et purification" xml:lang="fr"><term>Anticorps antiviraux</term><term>Anticorps monoclonaux</term><term>Anticorps neutralisants</term><term>Immunoglobuline G</term><term>Sous-type H1N1 du virus de la grippe A</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Enzyme-Linked Immunosorbent Assay</term><term>Immunotherapy</term></keywords><keywords scheme="MESH" qualifier="usage thérapeutique" xml:lang="fr"><term>Anticorps antiviraux</term><term>Anticorps monoclonaux</term><term>Anticorps neutralisants</term><term>Immunoglobuline G</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Animals</term><term>Antibody Affinity</term><term>Cross Reactions</term><term>Female</term><term>Humans</term><term>Mice</term><term>Mice, Inbred BALB C</term><term>Sensitivity and Specificity</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Affinité des anticorps</term><term>Animaux</term><term>Femelle</term><term>Humains</term><term>Immunothérapie</term><term>Réactions croisées</term><term>Sensibilité et spécificité</term><term>Souris</term><term>Souris de lignée BALB C</term><term>Sous-type H1N1 du virus de la grippe A</term><term>Test ELISA</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">The 2009 H1N1 influenza pandemic was a major international public health crisis which caused considerable morbidity and mortality worldwide. The goal of this study was to produce anti-H1 monoclonal antibodies (MAbs) for improving diagnostic immunological assays and to develop potential immunotherapeutics. Nine MAbs were produced after immunizing mice with recombinant hemagglutinin (HA) protein from A/California/06/09. Two spleenocyte myeloma fusions yielded 1588 hybridoma cultures. After screening the hybridoma culture supernatants for antibody reactivity to rHA, nine clones were selected for further characterization. Cross-reactivity studies of the anti-rHA antibodies against a panel of influenza viruses (H1-H16) revealed eight out of nine MAbs were specific to the pandemic H1 subtype, except for MAb F256G2sc1 which also cross-reacted with H5 subtype virus. All MAbs were of the IgG1κ isotype, except F256G2sc1 which was IgG2aκ. The anti-rHA MAbs had binding affinities to rHA that ranged from a K(D) (disassociation constant) of 1.34×10(-9)M (F255G7sc1) to the weakest affinity of 4.60×10(-8)M (F255G4sc1). Interestingly, in a plaque reduction neutralization assay, all MAbs except F255G3sc1 demonstrated neutralizing ability. Furthermore, all MAbs except F255G3sc1 and F255G9sc1 exhibited anti-hemagglutinin activity against pandemic H1N1 viruses, but not against classical North American swine influenza viruses of the same subtype. Immunofluorescence assay (IFA) demonstrated that all MAbs except F255G1sc1 and F255G3sc1 were able to detect 2009 pandemic H1N1 (2009) virus- infected MDCK cells. The MAbs were also evaluated for potential use in competitive ELISA (cELISA), and with the exception of F255G3sc1, all MAbs showed competitive activity with serum collected from pigs infected with pandemic H1N1 virus (2009). The developed MAbs have demonstrated utility as immunodiagnostic and research reagents, and their neutralizing capabilities also hold potential for designing antiviral drugs against pandemic influenza.</div></front></TEI><affiliations><list><country><li>Canada</li></country></list><tree><noCountry><name sortKey="Berhane, Yohannes" sort="Berhane, Yohannes" uniqKey="Berhane Y" first="Yohannes" last="Berhane">Yohannes Berhane</name><name sortKey="Corbett, Cindi R" sort="Corbett, Cindi R" uniqKey="Corbett C" first="Cindi R" last="Corbett">Cindi R. Corbett</name><name sortKey="Hole, Kate" sort="Hole, Kate" uniqKey="Hole K" first="Kate" last="Hole">Kate Hole</name><name sortKey="Leith, Marsha" sort="Leith, Marsha" uniqKey="Leith M" first="Marsha" last="Leith">Marsha Leith</name><name sortKey="Schmidt, Lisa" sort="Schmidt, Lisa" uniqKey="Schmidt L" first="Lisa" last="Schmidt">Lisa Schmidt</name><name sortKey="Tracz, Dobryan M" sort="Tracz, Dobryan M" uniqKey="Tracz D" first="Dobryan M" last="Tracz">Dobryan M. Tracz</name></noCountry><country name="Canada"><noRegion><name sortKey="Cabral, Teresa M" sort="Cabral, Teresa M" uniqKey="Cabral T" first="Teresa M" last="Cabral">Teresa M. Cabral</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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