Single amino acid substitutions in the severe acute respiratory syndrome coronavirus spike glycoprotein determine viral entry and immunogenicity of a major neutralizing domain
Identifieur interne : 004D63 ( Main/Exploration ); précédent : 004D62; suivant : 004D64Single amino acid substitutions in the severe acute respiratory syndrome coronavirus spike glycoprotein determine viral entry and immunogenicity of a major neutralizing domain
Auteurs : Christopher E. Yi [États-Unis] ; LEI BA [États-Unis] ; LINQI ZHANG [États-Unis] ; David D. Ho [États-Unis] ; ZHIWEI CHEN [États-Unis]Source :
- Journal of virology [ 0022-538X ] ; 2005.
Descripteurs français
- KwdFr :
- Animaux, Anticorps antiviraux (biosynthèse), Antigènes viraux (), Antigènes viraux (génétique), Délétion de séquence, Femelle, Glycoprotéine de spicule des coronavirus, Glycoprotéines membranaires (), Glycoprotéines membranaires (génétique), Glycoprotéines membranaires (immunologie), Glycoprotéines membranaires (physiologie), Humains, Lignée cellulaire, Microscopie électronique, Mutagenèse dirigée, Protéines de l'enveloppe virale (), Protéines de l'enveloppe virale (génétique), Protéines de l'enveloppe virale (immunologie), Protéines de l'enveloppe virale (physiologie), Souris, Souris de lignée BALB C, Structure tertiaire des protéines, Substitution d'acide aminé, Séquence d'acides aminés, Tests de neutralisation, Vaccins antiviraux (administration et posologie), Vaccins antiviraux (génétique), Vaccins à ADN (administration et posologie), Vaccins à ADN (génétique), Virulence (génétique), Virulence (immunologie), Virulence (physiologie), Virus du SRAS (génétique), Virus du SRAS (immunologie), Virus du SRAS (pathogénicité), Virus du SRAS (physiologie).
- MESH :
- administration et posologie : Vaccins antiviraux, Vaccins à ADN.
- biosynthèse : Anticorps antiviraux.
- génétique : Antigènes viraux, Glycoprotéines membranaires, Protéines de l'enveloppe virale, Vaccins antiviraux, Vaccins à ADN, Virulence, Virus du SRAS.
- immunologie : Glycoprotéines membranaires, Protéines de l'enveloppe virale, Virulence, Virus du SRAS.
- pathogénicité : Virus du SRAS.
- physiologie : Glycoprotéines membranaires, Protéines de l'enveloppe virale, Virulence, Virus du SRAS.
- Pascal (Inist)
- Animaux, Antigènes viraux, Coronavirus, Délétion de séquence, Femelle, Glycoprotéine, Glycoprotéine de spicule des coronavirus, Glycoprotéines membranaires, Humains, Immunogénicité, Lignée cellulaire, Microbiologie, Microscopie électronique, Mutagenèse dirigée, Protéines de l'enveloppe virale, Souris, Souris de lignée BALB C, Structure tertiaire des protéines, Substitution d'acide aminé, Séquence d'acides aminés, Tests de neutralisation, Virologie, Syndrome respiratoire aigu sévère.
English descriptors
- KwdEn :
- Amino Acid Sequence, Amino Acid Substitution, Animals, Antibodies, Viral (biosynthesis), Antigens, Viral (chemistry), Antigens, Viral (genetics), Cell Line, Coronavirus, Female, Glycoprotein, Humans, Immunogenicity, Membrane Glycoproteins (chemistry), Membrane Glycoproteins (genetics), Membrane Glycoproteins (immunology), Membrane Glycoproteins (physiology), Mice, Mice, Inbred BALB C, Microbiology, Microscopy, Electron, Mutagenesis, Site-Directed, Neutralization Tests, Protein Structure, Tertiary, SARS Virus (genetics), SARS Virus (immunology), SARS Virus (pathogenicity), SARS Virus (physiology), Sequence Deletion, Severe acute respiratory syndrome, Spike Glycoprotein, Coronavirus, Vaccines, DNA (administration & dosage), Vaccines, DNA (genetics), Viral Envelope Proteins (chemistry), Viral Envelope Proteins (genetics), Viral Envelope Proteins (immunology), Viral Envelope Proteins (physiology), Viral Vaccines (administration & dosage), Viral Vaccines (genetics), Virology, Virulence (genetics), Virulence (immunology), Virulence (physiology).
- MESH :
- chemical , administration & dosage : Vaccines, DNA, Viral Vaccines.
- chemical , biosynthesis : Antibodies, Viral.
- chemical , chemistry : Antigens, Viral, Membrane Glycoproteins, Viral Envelope Proteins.
- chemical , genetics : Antigens, Viral, Membrane Glycoproteins, Vaccines, DNA, Viral Envelope Proteins, Viral Vaccines.
- chemical , immunology : Membrane Glycoproteins, Viral Envelope Proteins.
- chemical , physiology : Membrane Glycoproteins, Viral Envelope Proteins.
- genetics : SARS Virus, Virulence.
- immunology : SARS Virus, Virulence.
- pathogenicity : SARS Virus.
- physiology : SARS Virus, Virulence.
- Amino Acid Sequence, Amino Acid Substitution, Animals, Cell Line, Female, Humans, Mice, Mice, Inbred BALB C, Microscopy, Electron, Mutagenesis, Site-Directed, Neutralization Tests, Protein Structure, Tertiary, Sequence Deletion, Spike Glycoprotein, Coronavirus.
Abstract
Neutralizing antibodies (NAbs) against severe acute respiratory syndrome (SARS) coronavirus (SARS-CoV) spike (S) glycoprotein confer protection to animals experimentally infected with the pathogenic virus. We and others previously demonstrated that a major mechanism for neutralizing SARS-CoV was through blocking the interaction between the S glycoprotein and the cellular receptor angiotensin-converting enzyme 2 (ACE2). In this study, we used in vivo electroporation DNA immunization and a pseudovirus-based assay to functionally evaluate immunogenicity and viral entry. We characterized the neutralization and viral entry determinants within the ACE2-binding domain of the S glycoprotein. The deletion of a positively charged region SA(422-463) abolished the capacity of the S glycoprotein to induce NAbs in mice vaccinated by in vivo DNA electroporation. Moreover, the SΔ(422-463) pseudovirus was unable to infect HEK293T-ACE2 cells. To determine the specific residues that contribute to related phenotypes, we replaced eight basic amino acids with alanine. We found that a single amino acid substitution (R441A) in the full-length S DNA vaccine failed to induce NAbs and abolished viral entry when pseudoviruses were generated. However, another substitution (R453A) abolished viral entry while retaining the capacity for inducing NAbs. The difference between R441A and R453A suggests that the determinants for immunogenicity and viral entry may not be identical. Our findings provide direct evidence that these basic residues are essential for immunogenicity of the major neutralizing domain and for viral entry. Our data have implications for the rational design of vaccine and antiviral agents as well as for understanding viral tropism.
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Affiliations:
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Ho</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Aaron Diamond AIDS Research Center, The Rockefeller University</s1><s2>New York, New York 10016</s2><s3>USA</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ></inist:fA14><country>États-Unis</country><wicri:noRegion>New York, New York 10016</wicri:noRegion></affiliation></author><author><name sortKey="Zhiwei Chen" sort="Zhiwei Chen" uniqKey="Zhiwei Chen" last="Zhiwei Chen">ZHIWEI CHEN</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Aaron Diamond AIDS Research Center, The Rockefeller University</s1><s2>New York, New York 10016</s2><s3>USA</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ></inist:fA14><country>États-Unis</country><wicri:noRegion>New York, New York 10016</wicri:noRegion></affiliation></author></analytic><series><title level="j" type="main">Journal of virology</title><title level="j" type="abbreviated">J. virol.</title><idno type="ISSN">0022-538X</idno><imprint><date when="2005">2005</date></imprint></series></biblStruct></sourceDesc><seriesStmt><title level="j" type="main">Journal of virology</title><title level="j" type="abbreviated">J. virol.</title><idno type="ISSN">0022-538X</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Amino Acid Sequence</term><term>Amino Acid Substitution</term><term>Animals</term><term>Antibodies, Viral (biosynthesis)</term><term>Antigens, Viral (chemistry)</term><term>Antigens, Viral (genetics)</term><term>Cell Line</term><term>Coronavirus</term><term>Female</term><term>Glycoprotein</term><term>Humans</term><term>Immunogenicity</term><term>Membrane Glycoproteins (chemistry)</term><term>Membrane Glycoproteins (genetics)</term><term>Membrane Glycoproteins (immunology)</term><term>Membrane Glycoproteins (physiology)</term><term>Mice</term><term>Mice, Inbred BALB C</term><term>Microbiology</term><term>Microscopy, Electron</term><term>Mutagenesis, Site-Directed</term><term>Neutralization Tests</term><term>Protein Structure, Tertiary</term><term>SARS Virus (genetics)</term><term>SARS Virus (immunology)</term><term>SARS Virus (pathogenicity)</term><term>SARS Virus (physiology)</term><term>Sequence Deletion</term><term>Severe acute respiratory syndrome</term><term>Spike Glycoprotein, Coronavirus</term><term>Vaccines, DNA (administration & dosage)</term><term>Vaccines, DNA (genetics)</term><term>Viral Envelope Proteins (chemistry)</term><term>Viral Envelope Proteins (genetics)</term><term>Viral Envelope Proteins (immunology)</term><term>Viral Envelope Proteins (physiology)</term><term>Viral Vaccines (administration & dosage)</term><term>Viral Vaccines (genetics)</term><term>Virology</term><term>Virulence (genetics)</term><term>Virulence (immunology)</term><term>Virulence (physiology)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Animaux</term><term>Anticorps antiviraux (biosynthèse)</term><term>Antigènes viraux ()</term><term>Antigènes viraux (génétique)</term><term>Délétion de séquence</term><term>Femelle</term><term>Glycoprotéine de spicule des coronavirus</term><term>Glycoprotéines membranaires ()</term><term>Glycoprotéines membranaires (génétique)</term><term>Glycoprotéines membranaires (immunologie)</term><term>Glycoprotéines membranaires (physiologie)</term><term>Humains</term><term>Lignée cellulaire</term><term>Microscopie électronique</term><term>Mutagenèse dirigée</term><term>Protéines de l'enveloppe virale ()</term><term>Protéines de l'enveloppe virale (génétique)</term><term>Protéines de l'enveloppe virale (immunologie)</term><term>Protéines de l'enveloppe virale (physiologie)</term><term>Souris</term><term>Souris de lignée BALB C</term><term>Structure tertiaire des protéines</term><term>Substitution d'acide aminé</term><term>Séquence d'acides aminés</term><term>Tests de neutralisation</term><term>Vaccins antiviraux (administration et posologie)</term><term>Vaccins antiviraux (génétique)</term><term>Vaccins à ADN (administration et posologie)</term><term>Vaccins à ADN (génétique)</term><term>Virulence (génétique)</term><term>Virulence (immunologie)</term><term>Virulence (physiologie)</term><term>Virus du SRAS (génétique)</term><term>Virus du SRAS (immunologie)</term><term>Virus du SRAS (pathogénicité)</term><term>Virus du SRAS (physiologie)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="administration & dosage" xml:lang="en"><term>Vaccines, DNA</term><term>Viral Vaccines</term></keywords><keywords scheme="MESH" type="chemical" qualifier="biosynthesis" xml:lang="en"><term>Antibodies, Viral</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Antigens, Viral</term><term>Membrane Glycoproteins</term><term>Viral Envelope Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Antigens, Viral</term><term>Membrane Glycoproteins</term><term>Vaccines, DNA</term><term>Viral Envelope Proteins</term><term>Viral Vaccines</term></keywords><keywords scheme="MESH" type="chemical" qualifier="immunology" xml:lang="en"><term>Membrane Glycoproteins</term><term>Viral Envelope Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="physiology" xml:lang="en"><term>Membrane Glycoproteins</term><term>Viral Envelope Proteins</term></keywords><keywords scheme="MESH" qualifier="administration et posologie" xml:lang="fr"><term>Vaccins antiviraux</term><term>Vaccins à ADN</term></keywords><keywords scheme="MESH" qualifier="biosynthèse" xml:lang="fr"><term>Anticorps antiviraux</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>SARS Virus</term><term>Virulence</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Antigènes viraux</term><term>Glycoprotéines membranaires</term><term>Protéines de l'enveloppe virale</term><term>Vaccins antiviraux</term><term>Vaccins à ADN</term><term>Virulence</term><term>Virus du SRAS</term></keywords><keywords scheme="MESH" qualifier="immunologie" xml:lang="fr"><term>Glycoprotéines membranaires</term><term>Protéines de l'enveloppe virale</term><term>Virulence</term><term>Virus du SRAS</term></keywords><keywords scheme="MESH" qualifier="immunology" xml:lang="en"><term>SARS Virus</term><term>Virulence</term></keywords><keywords scheme="MESH" qualifier="pathogenicity" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="pathogénicité" xml:lang="fr"><term>Virus du SRAS</term></keywords><keywords scheme="MESH" qualifier="physiologie" xml:lang="fr"><term>Glycoprotéines membranaires</term><term>Protéines de l'enveloppe virale</term><term>Virulence</term><term>Virus du SRAS</term></keywords><keywords scheme="MESH" qualifier="physiology" xml:lang="en"><term>SARS Virus</term><term>Virulence</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Amino Acid Sequence</term><term>Amino Acid Substitution</term><term>Animals</term><term>Cell Line</term><term>Female</term><term>Humans</term><term>Mice</term><term>Mice, Inbred BALB C</term><term>Microscopy, Electron</term><term>Mutagenesis, Site-Directed</term><term>Neutralization Tests</term><term>Protein Structure, Tertiary</term><term>Sequence Deletion</term><term>Spike Glycoprotein, Coronavirus</term></keywords><keywords scheme="Pascal" xml:lang="fr"><term>Animaux</term><term>Antigènes viraux</term><term>Coronavirus</term><term>Délétion de séquence</term><term>Femelle</term><term>Glycoprotéine</term><term>Glycoprotéine de spicule des coronavirus</term><term>Glycoprotéines membranaires</term><term>Humains</term><term>Immunogénicité</term><term>Lignée cellulaire</term><term>Microbiologie</term><term>Microscopie électronique</term><term>Mutagenèse dirigée</term><term>Protéines de l'enveloppe virale</term><term>Souris</term><term>Souris de lignée BALB C</term><term>Structure tertiaire des protéines</term><term>Substitution d'acide aminé</term><term>Séquence d'acides aminés</term><term>Tests de neutralisation</term><term>Virologie</term><term>Syndrome respiratoire aigu sévère</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Neutralizing antibodies (NAbs) against severe acute respiratory syndrome (SARS) coronavirus (SARS-CoV) spike (S) glycoprotein confer protection to animals experimentally infected with the pathogenic virus. We and others previously demonstrated that a major mechanism for neutralizing SARS-CoV was through blocking the interaction between the S glycoprotein and the cellular receptor angiotensin-converting enzyme 2 (ACE2). In this study, we used in vivo electroporation DNA immunization and a pseudovirus-based assay to functionally evaluate immunogenicity and viral entry. We characterized the neutralization and viral entry determinants within the ACE2-binding domain of the S glycoprotein. The deletion of a positively charged region SA(422-463) abolished the capacity of the S glycoprotein to induce NAbs in mice vaccinated by in vivo DNA electroporation. Moreover, the SΔ(422-463) pseudovirus was unable to infect HEK293T-ACE2 cells. To determine the specific residues that contribute to related phenotypes, we replaced eight basic amino acids with alanine. We found that a single amino acid substitution (R441A) in the full-length S DNA vaccine failed to induce NAbs and abolished viral entry when pseudoviruses were generated. However, another substitution (R453A) abolished viral entry while retaining the capacity for inducing NAbs. The difference between R441A and R453A suggests that the determinants for immunogenicity and viral entry may not be identical. Our findings provide direct evidence that these basic residues are essential for immunogenicity of the major neutralizing domain and for viral entry. Our data have implications for the rational design of vaccine and antiviral agents as well as for understanding viral tropism.</div></front></TEI><affiliations><list><country><li>États-Unis</li></country></list><tree><country name="États-Unis"><noRegion><name sortKey="Yi, Christopher E" sort="Yi, Christopher E" uniqKey="Yi C" first="Christopher E." last="Yi">Christopher E. Yi</name></noRegion><name sortKey="Ho, David D" sort="Ho, David D" uniqKey="Ho D" first="David D." last="Ho">David D. Ho</name><name sortKey="Lei Ba" sort="Lei Ba" uniqKey="Lei Ba" last="Lei Ba">LEI BA</name><name sortKey="Linqi Zhang" sort="Linqi Zhang" uniqKey="Linqi Zhang" last="Linqi Zhang">LINQI ZHANG</name><name sortKey="Zhiwei Chen" sort="Zhiwei Chen" uniqKey="Zhiwei Chen" last="Zhiwei Chen">ZHIWEI CHEN</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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