A severe acute respiratory syndrome coronavirus that lacks the E gene is attenuated in vitro and in vivo
Identifieur interne : 003B92 ( Main/Exploration ); précédent : 003B91; suivant : 003B93A severe acute respiratory syndrome coronavirus that lacks the E gene is attenuated in vitro and in vivo
Auteurs : Marta L. Dediego [Espagne] ; Enrique Alvarez [Espagne] ; Fernando Almazan [Espagne] ; Maria Teresa Rejas [Espagne] ; Elaine Lamirande [États-Unis] ; Anieanette Roberts [États-Unis] ; Wun-Ju Shieh [États-Unis] ; Sherif R. Zaki [États-Unis] ; Kanta Subbarao [États-Unis] ; Luis Enjuanes [Espagne]Source :
- Journal of virology [ 0022-538X ] ; 2007.
Descripteurs français
- Pascal (Inist)
English descriptors
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Abstract
A deletion mutant of severe acute respiratory syndrome coronavirus (SARS-CoV) has been engineered by deleting the structural E gene in an infectious cDNA clone that was constructed as a bacterial artificial chromosome (BAC). The recombinant virus lacking the E gene (rSARS-CoV-AE) was rescued in Vero E6 cells. The recovered deletion mutant grew in Vero E6, Huh-7, and CaCo-2 cells to titers 20-, 200-, and 200-fold lower than the recombinant wild-type virus, respectively, indicating that although the E protein has an effect on growth, it is not essential for virus replication. No differences in virion stability under a wide range of pH and temperature were detected between the deletion mutant and recombinant wild-type viruses. Although both viruses showed the same morphology by electron microscopy, the process of morphogenesis seemed to be less efficient with the defective virus than with the recombinant wild-type one. The rSARS-CoV-AE virus replicated to titers 100- to 1,000-fold lower than the recombinant wild-type virus in the upper and lower respiratory tract of hamsters, and the lower viral load was accompanied by less inflammation in the lungs of hamsters infected with rSARS-CoV-AE virus than with the recombinant wild-type virus. Therefore, the SARS-CoV that lacks the E gene is attenuated in hamsters, might be a safer research tool, and may be a good candidate for the development of a live attenuated SARS-CoV vaccine.
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Dediego</name><affiliation wicri:level="2"><inist:fA14 i1="01"><s1>Department of Molecular and Cell Biology, Centro Nacional de Biotecnología (CSIC), Campus Universidad Aut ónoma, Darwin 3, Cantoblanco</s1><s2>28049 Madrid</s2><s3>ESP</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>10 aut.</sZ></inist:fA14><country>Espagne</country><placeName><region nuts="2" type="communauté">Communauté de Madrid</region></placeName></affiliation></author><author><name sortKey="Alvarez, Enrique" sort="Alvarez, Enrique" uniqKey="Alvarez E" first="Enrique" last="Alvarez">Enrique Alvarez</name><affiliation wicri:level="2"><inist:fA14 i1="01"><s1>Department of Molecular and Cell Biology, Centro Nacional de Biotecnología (CSIC), Campus Universidad Aut ónoma, Darwin 3, Cantoblanco</s1><s2>28049 Madrid</s2><s3>ESP</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>10 aut.</sZ></inist:fA14><country>Espagne</country><placeName><region nuts="2" type="communauté">Communauté de Madrid</region></placeName></affiliation></author><author><name sortKey="Almazan, Fernando" sort="Almazan, Fernando" uniqKey="Almazan F" first="Fernando" last="Almazan">Fernando Almazan</name><affiliation wicri:level="2"><inist:fA14 i1="01"><s1>Department of Molecular and Cell Biology, Centro Nacional de Biotecnología (CSIC), Campus Universidad Aut ónoma, Darwin 3, Cantoblanco</s1><s2>28049 Madrid</s2><s3>ESP</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>10 aut.</sZ></inist:fA14><country>Espagne</country><placeName><region nuts="2" type="communauté">Communauté de Madrid</region></placeName></affiliation></author><author><name sortKey="Rejas, Maria Teresa" sort="Rejas, Maria Teresa" uniqKey="Rejas M" first="Maria Teresa" last="Rejas">Maria Teresa Rejas</name><affiliation wicri:level="2"><inist:fA14 i1="02"><s1>Centro de Biología Molecular (CSIC-UAM), Facultad de Ciencias, Campus Universidad Autónoma, Cantoblanco</s1><s2>28049 Madrid</s2><s3>ESP</s3><sZ>4 aut.</sZ></inist:fA14><country>Espagne</country><placeName><region nuts="2" type="communauté">Communauté de Madrid</region></placeName></affiliation></author><author><name sortKey="Lamirande, Elaine" sort="Lamirande, Elaine" uniqKey="Lamirande E" first="Elaine" last="Lamirande">Elaine Lamirande</name><affiliation wicri:level="1"><inist:fA14 i1="03"><s1>Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health</s1><s2>Bethesda, Maryland 208923</s2><s3>USA</s3><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>9 aut.</sZ></inist:fA14><country>États-Unis</country><wicri:noRegion>Bethesda, Maryland 208923</wicri:noRegion></affiliation></author><author><name sortKey="Roberts, Anieanette" sort="Roberts, Anieanette" uniqKey="Roberts A" first="Anieanette" last="Roberts">Anieanette Roberts</name><affiliation wicri:level="1"><inist:fA14 i1="03"><s1>Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health</s1><s2>Bethesda, Maryland 208923</s2><s3>USA</s3><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>9 aut.</sZ></inist:fA14><country>États-Unis</country><wicri:noRegion>Bethesda, Maryland 208923</wicri:noRegion></affiliation></author><author><name sortKey="Shieh, Wun Ju" sort="Shieh, Wun Ju" uniqKey="Shieh W" first="Wun-Ju" last="Shieh">Wun-Ju Shieh</name><affiliation wicri:level="1"><inist:fA14 i1="04"><s1>Infectious Disease Pathology Activity, National Center for Infectious Diseases, Centers for Disease Control and Prevention</s1><s2>Atlanta, Georgia 30333</s2><s3>USA</s3><sZ>7 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>États-Unis</country><wicri:noRegion>Atlanta, Georgia 30333</wicri:noRegion></affiliation></author><author><name sortKey="Zaki, Sherif R" sort="Zaki, Sherif R" uniqKey="Zaki S" first="Sherif R." last="Zaki">Sherif R. Zaki</name><affiliation wicri:level="1"><inist:fA14 i1="04"><s1>Infectious Disease Pathology Activity, National Center for Infectious Diseases, Centers for Disease Control and Prevention</s1><s2>Atlanta, Georgia 30333</s2><s3>USA</s3><sZ>7 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>États-Unis</country><wicri:noRegion>Atlanta, Georgia 30333</wicri:noRegion></affiliation></author><author><name sortKey="Subbarao, Kanta" sort="Subbarao, Kanta" uniqKey="Subbarao K" first="Kanta" last="Subbarao">Kanta Subbarao</name><affiliation wicri:level="1"><inist:fA14 i1="03"><s1>Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health</s1><s2>Bethesda, Maryland 208923</s2><s3>USA</s3><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>9 aut.</sZ></inist:fA14><country>États-Unis</country><wicri:noRegion>Bethesda, Maryland 208923</wicri:noRegion></affiliation></author><author><name sortKey="Enjuanes, Luis" sort="Enjuanes, Luis" uniqKey="Enjuanes L" first="Luis" last="Enjuanes">Luis Enjuanes</name><affiliation wicri:level="2"><inist:fA14 i1="01"><s1>Department of Molecular and Cell Biology, Centro Nacional de Biotecnología (CSIC), Campus Universidad Aut ónoma, Darwin 3, Cantoblanco</s1><s2>28049 Madrid</s2><s3>ESP</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>10 aut.</sZ></inist:fA14><country>Espagne</country><placeName><region nuts="2" type="communauté">Communauté de Madrid</region></placeName></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="2007">2007</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>Coronavirus</term><term>Gene</term><term>Severe acute respiratory syndrome</term><term>Virology</term></keywords><keywords scheme="Pascal" xml:lang="fr"><term>Coronavirus</term><term>Gène</term><term>Virologie</term><term>Syndrome respiratoire aigu sévère</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">A deletion mutant of severe acute respiratory syndrome coronavirus (SARS-CoV) has been engineered by deleting the structural E gene in an infectious cDNA clone that was constructed as a bacterial artificial chromosome (BAC). The recombinant virus lacking the E gene (rSARS-CoV-AE) was rescued in Vero E6 cells. The recovered deletion mutant grew in Vero E6, Huh-7, and CaCo-2 cells to titers 20-, 200-, and 200-fold lower than the recombinant wild-type virus, respectively, indicating that although the E protein has an effect on growth, it is not essential for virus replication. No differences in virion stability under a wide range of pH and temperature were detected between the deletion mutant and recombinant wild-type viruses. Although both viruses showed the same morphology by electron microscopy, the process of morphogenesis seemed to be less efficient with the defective virus than with the recombinant wild-type one. The rSARS-CoV-AE virus replicated to titers 100- to 1,000-fold lower than the recombinant wild-type virus in the upper and lower respiratory tract of hamsters, and the lower viral load was accompanied by less inflammation in the lungs of hamsters infected with rSARS-CoV-AE virus than with the recombinant wild-type virus. Therefore, the SARS-CoV that lacks the E gene is attenuated in hamsters, might be a safer research tool, and may be a good candidate for the development of a live attenuated SARS-CoV vaccine.</div></front></TEI><affiliations><list><country><li>Espagne</li><li>États-Unis</li></country><region><li>Communauté de Madrid</li></region></list><tree><country name="Espagne"><region name="Communauté de Madrid"><name sortKey="Dediego, Marta L" sort="Dediego, Marta L" uniqKey="Dediego M" first="Marta L." last="Dediego">Marta L. Dediego</name></region><name sortKey="Almazan, Fernando" sort="Almazan, Fernando" uniqKey="Almazan F" first="Fernando" last="Almazan">Fernando Almazan</name><name sortKey="Alvarez, Enrique" sort="Alvarez, Enrique" uniqKey="Alvarez E" first="Enrique" last="Alvarez">Enrique Alvarez</name><name sortKey="Enjuanes, Luis" sort="Enjuanes, Luis" uniqKey="Enjuanes L" first="Luis" last="Enjuanes">Luis Enjuanes</name><name sortKey="Rejas, Maria Teresa" sort="Rejas, Maria Teresa" uniqKey="Rejas M" first="Maria Teresa" last="Rejas">Maria Teresa Rejas</name></country><country name="États-Unis"><noRegion><name sortKey="Lamirande, Elaine" sort="Lamirande, Elaine" uniqKey="Lamirande E" first="Elaine" last="Lamirande">Elaine Lamirande</name></noRegion><name sortKey="Roberts, Anieanette" sort="Roberts, Anieanette" uniqKey="Roberts A" first="Anieanette" last="Roberts">Anieanette Roberts</name><name sortKey="Shieh, Wun Ju" sort="Shieh, Wun Ju" uniqKey="Shieh W" first="Wun-Ju" last="Shieh">Wun-Ju Shieh</name><name sortKey="Subbarao, Kanta" sort="Subbarao, Kanta" uniqKey="Subbarao K" first="Kanta" last="Subbarao">Kanta Subbarao</name><name sortKey="Zaki, Sherif R" sort="Zaki, Sherif R" uniqKey="Zaki S" first="Sherif R." last="Zaki">Sherif R. 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