Severe acute respiratory syndrome coronavirus spike protein expressed by attenuated vaccinia virus protectively immunizes mice

Himani Bisht, Anjeanette Roberts, Leatrice Vogel, Alexander Bukreyev, Peter L. Collins, Brian R. Murphy, Kanta Subbarao, Bernard Moss

Research output: Contribution to journalArticle

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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/S or 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-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 in vitro. 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.

Original languageEnglish (US)
Pages (from-to)6641-6646
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume101
Issue number17
DOIs
StatePublished - Apr 27 2004
Externally publishedYes

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Coronavirus Spike Glycoproteins
Severe Acute Respiratory Syndrome
Vaccinia virus
Coronavirus
Respiratory System
Antibodies
Epitopes
Vaccines
Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase
Glycoside Hydrolases
Protein S
Neutralizing Antibodies
Serum
Confocal Microscopy
Immunity

ASJC Scopus subject areas

  • Genetics
  • General

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Severe acute respiratory syndrome coronavirus spike protein expressed by attenuated vaccinia virus protectively immunizes mice. / Bisht, Himani; Roberts, Anjeanette; Vogel, Leatrice; Bukreyev, Alexander; Collins, Peter L.; Murphy, Brian R.; Subbarao, Kanta; Moss, Bernard.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 101, No. 17, 27.04.2004, p. 6641-6646.

Research output: Contribution to journalArticle

Bisht, Himani ; Roberts, Anjeanette ; Vogel, Leatrice ; Bukreyev, Alexander ; Collins, Peter L. ; Murphy, Brian R. ; Subbarao, Kanta ; Moss, Bernard. / Severe acute respiratory syndrome coronavirus spike protein expressed by attenuated vaccinia virus protectively immunizes mice. In: Proceedings of the National Academy of Sciences of the United States of America. 2004 ; Vol. 101, No. 17. pp. 6641-6646.
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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/S or 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-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 in vitro. 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 na{\"i}ve 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.",
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