title: Immunogenicity of a receptor-binding domain of SARS coronavirus spike protein in mice: implications for a subunit vaccine.
abstract: We studied the immunogenicity of an anti-SARS subunit vaccine comprised of the fragment of the SARS coronavirus (SARS-CoV) spike protein amino acids 318-510 (S318-510) containing the receptor-binding domain. The S protein fragment was purified from the culture supernatant of stably transformed HEK293T cells secreting a tagged version of the protein. The vaccine was given subcutaneously to 129S6/SvEv mice in saline, with alum adjuvant or with alum plus CpG oligodeoxynucleotides (ODN). Mice immunized with the adjuvanted antigen elicited strong antibody and cellular immune responses; furthermore, adding the CpG ODN to the alum resulted in increased IgG2a antibody titers and a higher number of INF-gamma-secreting murine splenocytes. Mice vaccinated with S318-510 deglycosylated by PNGase F (dgS318-510) showed a lower neutralizing antibody response but had similar numbers of INF-gamma-producing cells in the spleen. This finding suggests that carbohydrate is important for the immunogenicity of the S318-510 protein fragment and provide useful information for designing an effective and safe SARS subunit vaccine.
authors.affiliation: Vaccine and Infectious Disease Organization (VIDO), University of Saskatchewan, 120 Veterinary Road, Saskatoon, Sask., Canada S7N 5E3. zakhartchouk@sask.usask.ca
authors.forename: Alexander N
authors.initials: AN
authors.lastname: Zakhartchouk
authors.forename: Chetna
authors.initials: C
authors.lastname: Sharon
authors.forename: Malathy
authors.initials: M
authors.lastname: Satkunarajah
authors.forename: Thierry
authors.initials: T
authors.lastname: Auperin
authors.forename: Sathiyanarayanan
authors.initials: S
authors.lastname: Viswanathan
authors.forename: George
authors.initials: G
authors.lastname: Mutwiri
authors.forename: Martin
authors.initials: M
authors.lastname: Petric
authors.forename: Raymond H
authors.initials: RH
authors.lastname: See
authors.forename: Robert C
authors.initials: RC
authors.lastname: Brunham
authors.forename: B Brett
authors.initials: BB
authors.lastname: Finlay
authors.forename: Cheryl
authors.initials: C
authors.lastname: Cameron
authors.forename: David J
authors.initials: DJ
authors.lastname: Kelvin
authors.forename: Alan
authors.initials: A
authors.lastname: Cochrane
authors.forename: James M
authors.initials: JM
authors.lastname: Rini
authors.forename: Lorne A
authors.initials: LA
authors.lastname: Babiuk
mesh_terms: D000534:Alum Compounds; D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D002460:Cell Line; D005260:Female; D006801:Humans; D007371:Interferon-gamma / Q000096:biosynthesis; D008562:Membrane Glycoproteins / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D051379:Mice; D009838:Oligodeoxyribonucleotides; D011991:Receptors, Virus / Q000378:metabolism*; D045473:Severe acute respiratory syndrome-related coronavirus / Q000276:immunology*; D045169:Severe Acute Respiratory Syndrome / Q000276:immunology / Q000517:prevention & control*; D064370:Spike Glycoprotein, Coronavirus; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D014765:Viral Vaccines / Q000008:administration & dosage / Q000276:immunology*
chemical_list: D000534:Alum Compounds; D000914:Antibodies, Viral; D008562:Membrane Glycoproteins; D009838:Oligodeoxyribonucleotides; D011991:Receptors, Virus; D064370:Spike Glycoprotein, Coronavirus; D014759:Viral Envelope Proteins; D014765:Viral Vaccines; C578557:spike glycoprotein, SARS-CoV; C578553:spike protein, mouse hepatitis virus; C041524:aluminum sulfate; D007371:Interferon-gamma