A double-inactivated whole virus candidate SARS coronavirus vaccine stimulates neutralising and protective antibody responses - PubMed (original) (raw)

. 2006 Jan 30;24(5):652-61.

doi: 10.1016/j.vaccine.2005.08.055. Epub 2005 Aug 26.

Otfried Kistner, Helga Savidis-Dacho, Elisabeth Hitter, Brian Crowe, Marijan Gerencer, Peter Brühl, Leopold Grillberger, Manfred Reiter, Christa Tauer, Wolfgang Mundt, P Noel Barrett

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A double-inactivated whole virus candidate SARS coronavirus vaccine stimulates neutralising and protective antibody responses

Martin Spruth et al. Vaccine. 2006.

Abstract

A double-inactivated, candidate whole virus vaccine against severe acute respiratory syndrome associated coronavirus (SARS-CoV) was developed and manufactured at large scale using fermenter cultures of serum protein free Vero cells. A two step inactivation procedure involving sequential formaldehyde and U.V. inactivation was utilised in order to ensure an extremely high safety margin with respect to residual infectivity. The immunogenicity of this double-inactivated vaccine was characterised in the mouse model. Mice that were immunised twice with the candidate SARS-CoV vaccine developed high antibody titres against the SARS-CoV spike protein and high levels of neutralising antibodies. The use of the adjuvant Al(OH)3 had only a minor effect on the immunogenicity of the vaccine. In addition, cell mediated immunity as measured by interferon-gamma and interleukin-4 stimulation, was elicited by vaccination. Moreover, the vaccine confers protective immunity as demonstrated by prevention of SARS-CoV replication in the respiratory tract of mice after intranasal challenge with SARS-CoV. Protection of mice was correlated to antibody titre against the SARS-CoV S protein and neutralising antibody titre.

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Figures

Fig. 1

Fig. 1

Electrophoretic and Western blot analysis of purified SARS-CoV candidate vaccine. Lysate of SARS-CoV infected Vero SF cells (lane 1) or two lots of purified vaccine (lanes 2 and 3) were separated by SDS-PAGE. Western blot analysis was performed with antibodies specific for the Spike-protein (A) or the Nucleocapsid-protein (B) of SARS-CoV; protein staining using colloidal gold is shown in (C).

Fig. 2

Fig. 2

Electron micrograph (187,000-fold magnification) of purified inactivated SARS-CoV candidate vaccine after staining with uranyl acetate. Spikes formed by S protein project from the viral surface.

Fig. 3

Fig. 3

S-specific and neutralising antibody response against SARS-CoV vaccine of CD1 mice after booster immunisation. The S-specific antibody titre at week 4 (A) or 12 (C), respectively, was determined by ELISA. Neutralising antibody response of CD1 mice at week 5 (B) or week 13 (D), respectively, was analysed by a micro neutralisation assay. Geometric mean titres of groups that received the non-adjuvanted vaccine (grey bars), or vaccine adjuvanted with 0.05% (black bars) or 0.2% aluminium hydroxide (open bars) are presented (n = 10 for antibody titres; n = 4 for neutralising titres); (formula image) no neutralising antibodies detectable (due to limited amount of serum in some instances the detection limit of the assay was at 1:57 or 1:71).

Fig. 4

Fig. 4

Cellular immune response to vaccination with SARS-CoV vaccine. Interferon-γ (A) and interleukin-4 responses (B) were determined by ELISPOT assays: spleen cells of naive Balb/C mice, or spleen cell obtained from of immunised Balb/C mice at day 21 or 35 after primary immunisation were stimulated with a SARS-CoV vaccine preparation, recombinant S protein or a Vero mock preparation.

References

    1. Peiris J.S.M., Lai S.T., Poon L.L.M., Guan Y., Yam L.Y.C., Lim W. Coronavirus as a possible cause of severe acute respiratory syndrome. Lancet. 2003;361(9366):1319–1325. - PMC - PubMed
    1. Drosten C., Gunther S., Preiser W., van der Werf S., Brodt H.R., Becker S. Identification of a novel coronavirus in patients with severe acute respiratory syndrome. N Engl J Med. 2003;318(20):1967–1976. - PubMed
    1. Ksiazek T.G., Erdman D., Goldsmith C.S., Zaki S.R., Peret T., Emery S. A novel coronavirus associated with severe acute respiratory syndrome. N Engl J Med. 2003;348(20):1953–1966. - PubMed
    1. Guan Y., Zheng B.J., He Y.Q., Liu X.L., Zhuang Z.X., Cheung C.L. Isolation and characterization of virus related to the SARS coronavirus from animals in southern China. Science. 2003;302:276–278. - PubMed
    1. Ellis R.W. Technologies for making new vaccines. In: Plotkin S.A., Orenstein WA., editors. Vaccines. 4th ed. Elsevier; 2004. pp. 1177–1197.

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