Engineering a Replication-Competent, Propagation-Defective Middle East Respiratory Syndrome Coronavirus as a Vaccine Candidate
Identifieur interne : 002001 ( Main/Exploration ); précédent : 002000; suivant : 002002Engineering a Replication-Competent, Propagation-Defective Middle East Respiratory Syndrome Coronavirus as a Vaccine Candidate
Auteurs : Fernando Almazán ; Marta L. Dediego ; Isabel Sola ; Sonia Zu Iga ; Jose L. Nieto-Torres ; Silvia Marquez-Jurado ; German Andrés ; Luis EnjuanesSource :
- mBio [ 2150-7511 ] ; 2013.
Descripteurs français
- KwdFr :
- Coronaviridae (génétique), Coronaviridae (immunologie), Coronaviridae (physiologie), Génie génétique, Humains, Infections à Coronaviridae (virologie), Lignée cellulaire, Protéines virales (génétique), Protéines virales (immunologie), Réplication virale, Vaccins antiviraux (génétique), Vaccins antiviraux (immunologie).
- MESH :
- génétique : Coronaviridae, Protéines virales, Vaccins antiviraux.
- immunologie : Coronaviridae, Protéines virales, Vaccins antiviraux.
- physiologie : Coronaviridae.
- virologie : Infections à Coronaviridae.
- Génie génétique, Humains, Lignée cellulaire, Réplication virale.
English descriptors
- KwdEn :
- MESH :
- chemical , genetics : Viral Proteins, Viral Vaccines.
- genetics : Coronaviridae.
- immunology : Coronaviridae, Viral Proteins, Viral Vaccines.
- physiology : Coronaviridae.
- virology : Coronaviridae Infections.
- Cell Line, Genetic Engineering, Humans, Virus Replication.
Abstract
Middle East respiratory syndrome coronavirus (MERS-CoV) is an emerging coronavirus infecting humans that is associated with acute pneumonia, occasional renal failure, and a high mortality rate and is considered a threat to public health. The construction of a full-length infectious cDNA clone of the MERS-CoV genome in a bacterial artificial chromosome is reported here, providing a reverse genetics system to study the molecular biology of the virus and to develop attenuated viruses as vaccine candidates. Following transfection with the cDNA clone, infectious virus was rescued in both Vero A66 and Huh-7 cells. Recombinant MERS-CoVs (rMERS-CoVs) lacking the accessory genes 3, 4a, 4b, and 5 were successfully rescued from cDNA clones with these genes deleted. The mutant viruses presented growth kinetics similar to those of the wild-type virus, indicating that accessory genes were not essential for MERS-CoV replication in cell cultures. In contrast, an engineered mutant virus lacking the structural E protein (rMERS-CoV-ΔE) was not successfully rescued, since viral infectivity was lost at early passages. Interestingly, the rMERS-CoV-ΔE genome replicated after cDNA clone was transfected into cells. The infectious virus was rescued and propagated in cells expressing the E protein in
Url:
DOI: 10.1128/mBio.00650-13
PubMed: 24023385
PubMed Central: 3774192
Affiliations:
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Le document en format XML
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Dediego</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Sola, Isabel" sort="Sola, Isabel" uniqKey="Sola I" first="Isabel" last="Sola">Isabel Sola</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Zu Iga, Sonia" sort="Zu Iga, Sonia" uniqKey="Zu Iga S" first="Sonia" last="Zu Iga">Sonia Zu Iga</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Nieto Torres, Jose L" sort="Nieto Torres, Jose L" uniqKey="Nieto Torres J" first="Jose L." last="Nieto-Torres">Jose L. Nieto-Torres</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Marquez Jurado, Silvia" sort="Marquez Jurado, Silvia" uniqKey="Marquez Jurado S" first="Silvia" last="Marquez-Jurado">Silvia Marquez-Jurado</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Andres, German" sort="Andres, German" uniqKey="Andres G" first="German" last="Andrés">German Andrés</name><affiliation><nlm:aff id="aff2"></nlm:aff></affiliation></author><author><name sortKey="Enjuanes, Luis" sort="Enjuanes, Luis" uniqKey="Enjuanes L" first="Luis" last="Enjuanes">Luis Enjuanes</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author></analytic><series><title level="j">mBio</title><idno type="eISSN">2150-7511</idno><imprint><date when="2013">2013</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Cell Line</term><term>Coronaviridae (genetics)</term><term>Coronaviridae (immunology)</term><term>Coronaviridae (physiology)</term><term>Coronaviridae Infections (virology)</term><term>Genetic Engineering</term><term>Humans</term><term>Viral Proteins (genetics)</term><term>Viral Proteins (immunology)</term><term>Viral Vaccines (genetics)</term><term>Viral Vaccines (immunology)</term><term>Virus Replication</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Coronaviridae (génétique)</term><term>Coronaviridae (immunologie)</term><term>Coronaviridae (physiologie)</term><term>Génie génétique</term><term>Humains</term><term>Infections à Coronaviridae (virologie)</term><term>Lignée cellulaire</term><term>Protéines virales (génétique)</term><term>Protéines virales (immunologie)</term><term>Réplication virale</term><term>Vaccins antiviraux (génétique)</term><term>Vaccins antiviraux (immunologie)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Viral Proteins</term><term>Viral Vaccines</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Coronaviridae</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Coronaviridae</term><term>Protéines virales</term><term>Vaccins antiviraux</term></keywords><keywords scheme="MESH" qualifier="immunologie" xml:lang="fr"><term>Coronaviridae</term><term>Protéines virales</term><term>Vaccins antiviraux</term></keywords><keywords scheme="MESH" qualifier="immunology" xml:lang="en"><term>Coronaviridae</term><term>Viral Proteins</term><term>Viral Vaccines</term></keywords><keywords scheme="MESH" qualifier="physiologie" xml:lang="fr"><term>Coronaviridae</term></keywords><keywords scheme="MESH" qualifier="physiology" xml:lang="en"><term>Coronaviridae</term></keywords><keywords scheme="MESH" qualifier="virologie" xml:lang="fr"><term>Infections à Coronaviridae</term></keywords><keywords scheme="MESH" qualifier="virology" xml:lang="en"><term>Coronaviridae Infections</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Cell Line</term><term>Genetic Engineering</term><term>Humans</term><term>Virus Replication</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Génie génétique</term><term>Humains</term><term>Lignée cellulaire</term><term>Réplication virale</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><title>ABSTRACT</title><p>Middle East respiratory syndrome coronavirus (MERS-CoV) is an emerging coronavirus infecting humans that is associated with acute pneumonia, occasional renal failure, and a high mortality rate and is considered a threat to public health. The construction of a full-length infectious cDNA clone of the MERS-CoV genome in a bacterial artificial chromosome is reported here, providing a reverse genetics system to study the molecular biology of the virus and to develop attenuated viruses as vaccine candidates. Following transfection with the cDNA clone, infectious virus was rescued in both Vero A66 and Huh-7 cells. Recombinant MERS-CoVs (rMERS-CoVs) lacking the accessory genes 3, 4a, 4b, and 5 were successfully rescued from cDNA clones with these genes deleted. The mutant viruses presented growth kinetics similar to those of the wild-type virus, indicating that accessory genes were not essential for MERS-CoV replication in cell cultures. In contrast, an engineered mutant virus lacking the structural E protein (rMERS-CoV-ΔE) was not successfully rescued, since viral infectivity was lost at early passages. Interestingly, the rMERS-CoV-ΔE genome replicated after cDNA clone was transfected into cells. The infectious virus was rescued and propagated in cells expressing the E protein in <italic>trans</italic>, indicating that this virus was replication competent and propagation defective. Therefore, the rMERS-CoV-ΔE mutant virus is potentially a safe and promising vaccine candidate to prevent MERS-CoV infection.</p><p><bold>IMPORTANCE </bold> Since the emergence of MERS-CoV in the Arabian Peninsula during the summer of 2012, it has already spread to 10 different countries, infecting around 94 persons and showing a mortality rate higher than 50%. This article describes the development of the first reverse genetics system for MERS-CoV, based on the construction of an infectious cDNA clone inserted into a bacterial artificial chromosome. Using this system, a collection of rMERS-CoV deletion mutants has been generated. Interestingly, one of the mutants with the E gene deleted was a replication-competent, propagation-defective virus that could only be grown in the laboratory by providing E protein in <italic>trans</italic>, whereas it would only survive a single virus infection cycle <italic>in vivo</italic>. This virus constitutes a vaccine candidate that may represent a balance between safety and efficacy for the induction of mucosal immunity, which is needed to prevent MERS-CoV infection.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Danielsson, N" uniqKey="Danielsson N">N Danielsson</name></author><author><name sortKey="Catchpole, M" uniqKey="Catchpole M">M Catchpole</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Zaki, Am" uniqKey="Zaki A">AM Zaki</name></author><author><name sortKey="Van Boheemen, S" uniqKey="Van Boheemen S">S van Boheemen</name></author><author><name sortKey="Bestebroer, Tm" uniqKey="Bestebroer T">TM Bestebroer</name></author><author><name sortKey="Osterhaus, Ad" uniqKey="Osterhaus A">AD Osterhaus</name></author><author><name sortKey="Fouchier, Ra" uniqKey="Fouchier R">RA 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Dediego</name><name sortKey="Enjuanes, Luis" sort="Enjuanes, Luis" uniqKey="Enjuanes L" first="Luis" last="Enjuanes">Luis Enjuanes</name><name sortKey="Marquez Jurado, Silvia" sort="Marquez Jurado, Silvia" uniqKey="Marquez Jurado S" first="Silvia" last="Marquez-Jurado">Silvia Marquez-Jurado</name><name sortKey="Nieto Torres, Jose L" sort="Nieto Torres, Jose L" uniqKey="Nieto Torres J" first="Jose L." last="Nieto-Torres">Jose L. Nieto-Torres</name><name sortKey="Sola, Isabel" sort="Sola, Isabel" uniqKey="Sola I" first="Isabel" last="Sola">Isabel Sola</name><name sortKey="Zu Iga, Sonia" sort="Zu Iga, Sonia" uniqKey="Zu Iga S" first="Sonia" last="Zu Iga">Sonia Zu Iga</name></noCountry></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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|texte= Engineering a Replication-Competent, Propagation-Defective Middle East Respiratory Syndrome Coronavirus as a Vaccine Candidate
}}
Pour générer des pages wiki
HfdIndexSelect -h $EXPLOR_AREA/Data/Main/Exploration/RBID.i -Sk "pubmed:24023385" \
| HfdSelect -Kh $EXPLOR_AREA/Data/Main/Exploration/biblio.hfd \
| NlmPubMed2Wicri -a MersV1
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