Severe acute respiratory syndrome coronavirus spike protein expressed by attenuated vaccinia virus protectively immunizes mice
Identifieur interne : 000744 ( Pmc/Curation ); précédent : 000743; suivant : 000745Severe acute respiratory syndrome coronavirus spike protein expressed by attenuated vaccinia virus protectively immunizes mice
Auteurs : Himani Bisht ; Anjeanette Roberts [États-Unis] ; Leatrice Vogel [États-Unis] ; Alexander Bukreyev [États-Unis] ; Peter L. Collins [États-Unis] ; Brian R. Murphy [États-Unis] ; Kanta Subbarao [États-Unis] ; Bernard MossSource :
- Proceedings of the National Academy of Sciences of the United States of America [ 0027-8424 ] ; 2004.
Abstract
The spike protein (S), a membrane component of severe acute respiratory syndrome coronavirus (SARS-CoV) is anticipated to be an important component of candidate vaccines. We constructed recombinant forms of the highly attenuated modified vaccinia virus Ankara (MVA) containing the gene encoding full-length SARS-CoV S with and without a C-terminal epitope tag called MVA/S-HA and MVA/S, respectively. Cells infected with MVA/Sor MVA/S-HA synthesized a 200-kDa protein, which was recognized by antibody raised against a synthetic peptide of SARS-CoV S or the epitope tag in Western blot analyses. Further studies indicated that S was N
Url:
DOI: 10.1073/pnas.0401939101
PubMed: 15096611
PubMed Central: 404098
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<front><div type="abstract" xml:lang="en"><p>The spike protein (S), a membrane component of severe acute respiratory syndrome coronavirus (SARS-CoV) is anticipated to be an important component of candidate vaccines. We constructed recombinant forms of the highly attenuated modified vaccinia virus Ankara (MVA) containing the gene encoding full-length SARS-CoV S with and without a C-terminal epitope tag called MVA/S-HA and MVA/S, respectively. Cells infected with MVA/Sor MVA/S-HA synthesized a 200-kDa protein, which was recognized by antibody raised against a synthetic peptide of SARS-CoV S or the epitope tag in Western blot analyses. Further studies indicated that S was N<italic>-</italic>
glycosylated and migrated in SDS polyacrylamide gels with an apparent mass of ≈160 kDa after treatment with peptide N-glycosidase F. The acquisition of resistance to endoglycosidase H indicated trafficking of S to the medial Golgi compartment, and confocal microscopy showed that S was transported to the cell surface. Intranasal or intramuscular inoculations of BALB/c mice with MVA/S produced serum antibodies that recognized the SARS S in ELISA and neutralized SARS-CoV <italic>in vitro</italic>
. Moreover, MVA/S administered by either route elicited protective immunity, as shown by reduced titers of SARS-CoV in the upper and lower respiratory tracts of mice after challenge. Passive transfer of serum from mice immunized with MVA/S to naïve mice also reduced the replication of SARS-CoV in the respiratory tract after challenge, demonstrating a role for antibody to S in protection. The attenuated nature of MVA and the ability of MVA/S to induce neutralizing antibody that protects mice support further development of this candidate vaccine.</p>
</div>
</front>
</TEI>
<pmc article-type="research-article"><pmc-comment>The publisher of this article does not allow downloading of the full text in XML form.</pmc-comment>
<front><journal-meta><journal-id journal-id-type="nlm-ta">Proc Natl Acad Sci U S A</journal-id>
<journal-id journal-id-type="publisher-id">pnas</journal-id>
<journal-title>Proceedings of the National Academy of Sciences of the United States of America</journal-title>
<issn pub-type="ppub">0027-8424</issn>
<issn pub-type="epub">1091-6490</issn>
<publisher><publisher-name>National Academy of Sciences</publisher-name>
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<article-id pub-id-type="doi">10.1073/pnas.0401939101</article-id>
<article-categories><subj-group subj-group-type="heading"><subject>Biological Sciences</subject>
<subj-group><subject>Medical Sciences</subject>
</subj-group>
</subj-group>
</article-categories>
<title-group><article-title>Severe acute respiratory syndrome coronavirus spike protein expressed by attenuated vaccinia virus protectively immunizes mice</article-title>
</title-group>
<contrib-group><contrib contrib-type="author"><name><surname>Bisht</surname>
<given-names>Himani</given-names>
</name>
<xref ref-type="aff" rid="N0x9c4d208.0xa0c0df8">*</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Roberts</surname>
<given-names>Anjeanette</given-names>
</name>
<xref ref-type="aff" rid="N0x9c4d208.0xa0c0df8">†</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Vogel</surname>
<given-names>Leatrice</given-names>
</name>
<xref ref-type="aff" rid="N0x9c4d208.0xa0c0df8">†</xref>
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<contrib contrib-type="author"><name><surname>Bukreyev</surname>
<given-names>Alexander</given-names>
</name>
<xref ref-type="aff" rid="N0x9c4d208.0xa0c0df8">†</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Collins</surname>
<given-names>Peter L.</given-names>
</name>
<xref ref-type="aff" rid="N0x9c4d208.0xa0c0df8">†</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Murphy</surname>
<given-names>Brian R.</given-names>
</name>
<xref ref-type="aff" rid="N0x9c4d208.0xa0c0df8">†</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Subbarao</surname>
<given-names>Kanta</given-names>
</name>
<xref ref-type="aff" rid="N0x9c4d208.0xa0c0df8">†</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Moss</surname>
<given-names>Bernard</given-names>
</name>
<xref ref-type="aff" rid="N0x9c4d208.0xa0c0df8">*</xref>
<xref ref-type="corresp" rid="cor1">‡</xref>
</contrib>
</contrib-group>
<aff id="N0x9c4d208.0xa0c0df8"><label>*</label>
Laboratory of Viral Diseases and<label>†</label>
Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892</aff>
<author-notes><fn id="cor1"><label>‡</label>
<p> To whom correspondence should be addressed. E-mail: <email>bmoss@niaid.nih.gov</email>
. </p>
</fn>
<fn><p>Contributed by Bernard Moss, March 19, 2004</p>
</fn>
</author-notes>
<pub-date pub-type="ppub"><day>27</day>
<month>4</month>
<year>2004</year>
</pub-date>
<pub-date pub-type="epub"><day>19</day>
<month>4</month>
<year>2004</year>
</pub-date>
<volume>101</volume>
<issue>17</issue>
<fpage>6641</fpage>
<lpage>6646</lpage>
<copyright-statement>Copyright © 2004, The National Academy of Sciences</copyright-statement>
<copyright-year>2004</copyright-year>
<abstract><p>The spike protein (S), a membrane component of severe acute respiratory syndrome coronavirus (SARS-CoV) is anticipated to be an important component of candidate vaccines. We constructed recombinant forms of the highly attenuated modified vaccinia virus Ankara (MVA) containing the gene encoding full-length SARS-CoV S with and without a C-terminal epitope tag called MVA/S-HA and MVA/S, respectively. Cells infected with MVA/Sor MVA/S-HA synthesized a 200-kDa protein, which was recognized by antibody raised against a synthetic peptide of SARS-CoV S or the epitope tag in Western blot analyses. Further studies indicated that S was N<italic>-</italic>
glycosylated and migrated in SDS polyacrylamide gels with an apparent mass of ≈160 kDa after treatment with peptide N-glycosidase F. The acquisition of resistance to endoglycosidase H indicated trafficking of S to the medial Golgi compartment, and confocal microscopy showed that S was transported to the cell surface. Intranasal or intramuscular inoculations of BALB/c mice with MVA/S produced serum antibodies that recognized the SARS S in ELISA and neutralized SARS-CoV <italic>in vitro</italic>
. Moreover, MVA/S administered by either route elicited protective immunity, as shown by reduced titers of SARS-CoV in the upper and lower respiratory tracts of mice after challenge. Passive transfer of serum from mice immunized with MVA/S to naïve mice also reduced the replication of SARS-CoV in the respiratory tract after challenge, demonstrating a role for antibody to S in protection. The attenuated nature of MVA and the ability of MVA/S to induce neutralizing antibody that protects mice support further development of this candidate vaccine.</p>
</abstract>
</article-meta>
<notes><fn-group><fn><p>Abbreviations: SARS, severe acute respiratory syndrome; CoV, coronavirus; S, spike protein; CEF, chicken embryo fibroblast; MVA, modified vaccinia virus Ankara; pfu, plaque forming unit; HA, hemagglutinin; endo, endoglycosidase; PNGase, peptide N-glycosidase; i.n. intranasally; TCID<sub>50</sub>
, tissue culture 50% infective dose.</p>
</fn>
</fn-group>
</notes>
</front>
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