The expression and antigenicity of a truncated spike-nucleocapsid fusion protein of severe acute respiratory syndrome-associated coronavirus

Feng Mu, Dongsheng Niu, Jingsong Mu, Bo He, Weiguo Han, Baoxing Fan, Shengyong Huang, Yan Qiu, Bo You, Weijun Chen

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Background. In the absence of effective drugs, controlling SARS relies on the rapid identification of cases and appropriate management of the close contacts, or effective vaccines for SARS. Therefore, developing specific and sensitive laboratory tests for SARS as well as effective vaccines are necessary for national authorities. Results. Genes encoding truncated nucleocapsid (N) and spike (S) proteins of SARSCoV were cloned into the expression vector pQE30 and fusionally expressed in Escherichia coli M15. The fusion protein was analyzed for reactivity with SARS patients' sera and with anti-sera against the two human coronaviruses HCoV 229E and HCoV OC43 by ELISA, IFA and immunoblot assays. Furthermore, to evaluate the antigen-specific humoral antibody and T-cell responses in mice, the fusion protein was injected into 6-week-old BALB/c mice and a neutralization test as well as a T-cell analysis was performed. To evaluate the antiviral efficacy of immunization, BALB/c mice were challenged intranasally with SARSCoV at day 33 post injection and viral loads were determined by fluorescent quantitative RT-PCR. Serological results showed that the diagnostic sensitivity and specificity of the truncated S-N fusion protein derived the SARS virus were > 99% (457/460) and 100.00% (650/650), respectively. Furthermore there was no cross-reactivity with other two human coronaviruses. High titers of antibodies to SRASCoV appeared in the immunized mice and the neutralization test showed that antibodies to the fusion protein could inhibit SARSCoV. The T cell proliferation showed that the fusion protein could induce an antigen-specific T-cell response. Fluorescent quantitative RT-PCR showed that BALB/c mice challenged intranasally with SARSCoV at day 33 post injection were completely protected from virus replication. Conclusion. The truncated S-N fusion protein is a suitable immunodiagnostic antigen and vaccine candidate.

Original languageEnglish (US)
Article number207
JournalBMC Microbiology
Volume8
DOIs
StatePublished - 2008
Externally publishedYes

Fingerprint

Nucleocapsid Proteins
Severe Acute Respiratory Syndrome
Coronavirus
T-Lymphocytes
Neutralization Tests
Vaccines
Proteins
Antigens
Antibodies
Human Coronavirus 229E
SARS Virus
Nucleocapsid
Polymerase Chain Reaction
Injections
Protein S
Case Management
Virus Replication
Viral Load
Serum
Antiviral Agents

ASJC Scopus subject areas

  • Microbiology (medical)
  • Microbiology

Cite this

The expression and antigenicity of a truncated spike-nucleocapsid fusion protein of severe acute respiratory syndrome-associated coronavirus. / Mu, Feng; Niu, Dongsheng; Mu, Jingsong; He, Bo; Han, Weiguo; Fan, Baoxing; Huang, Shengyong; Qiu, Yan; You, Bo; Chen, Weijun.

In: BMC Microbiology, Vol. 8, 207, 2008.

Research output: Contribution to journalArticle

Mu, Feng ; Niu, Dongsheng ; Mu, Jingsong ; He, Bo ; Han, Weiguo ; Fan, Baoxing ; Huang, Shengyong ; Qiu, Yan ; You, Bo ; Chen, Weijun. / The expression and antigenicity of a truncated spike-nucleocapsid fusion protein of severe acute respiratory syndrome-associated coronavirus. In: BMC Microbiology. 2008 ; Vol. 8.
@article{00356f947352474c8000936d7c285cf2,
title = "The expression and antigenicity of a truncated spike-nucleocapsid fusion protein of severe acute respiratory syndrome-associated coronavirus",
abstract = "Background. In the absence of effective drugs, controlling SARS relies on the rapid identification of cases and appropriate management of the close contacts, or effective vaccines for SARS. Therefore, developing specific and sensitive laboratory tests for SARS as well as effective vaccines are necessary for national authorities. Results. Genes encoding truncated nucleocapsid (N) and spike (S) proteins of SARSCoV were cloned into the expression vector pQE30 and fusionally expressed in Escherichia coli M15. The fusion protein was analyzed for reactivity with SARS patients' sera and with anti-sera against the two human coronaviruses HCoV 229E and HCoV OC43 by ELISA, IFA and immunoblot assays. Furthermore, to evaluate the antigen-specific humoral antibody and T-cell responses in mice, the fusion protein was injected into 6-week-old BALB/c mice and a neutralization test as well as a T-cell analysis was performed. To evaluate the antiviral efficacy of immunization, BALB/c mice were challenged intranasally with SARSCoV at day 33 post injection and viral loads were determined by fluorescent quantitative RT-PCR. Serological results showed that the diagnostic sensitivity and specificity of the truncated S-N fusion protein derived the SARS virus were > 99{\%} (457/460) and 100.00{\%} (650/650), respectively. Furthermore there was no cross-reactivity with other two human coronaviruses. High titers of antibodies to SRASCoV appeared in the immunized mice and the neutralization test showed that antibodies to the fusion protein could inhibit SARSCoV. The T cell proliferation showed that the fusion protein could induce an antigen-specific T-cell response. Fluorescent quantitative RT-PCR showed that BALB/c mice challenged intranasally with SARSCoV at day 33 post injection were completely protected from virus replication. Conclusion. The truncated S-N fusion protein is a suitable immunodiagnostic antigen and vaccine candidate.",
author = "Feng Mu and Dongsheng Niu and Jingsong Mu and Bo He and Weiguo Han and Baoxing Fan and Shengyong Huang and Yan Qiu and Bo You and Weijun Chen",
year = "2008",
doi = "10.1186/1471-2180-8-207",
language = "English (US)",
volume = "8",
journal = "BMC Microbiology",
issn = "1471-2180",
publisher = "BioMed Central",

}

TY - JOUR

T1 - The expression and antigenicity of a truncated spike-nucleocapsid fusion protein of severe acute respiratory syndrome-associated coronavirus

AU - Mu, Feng

AU - Niu, Dongsheng

AU - Mu, Jingsong

AU - He, Bo

AU - Han, Weiguo

AU - Fan, Baoxing

AU - Huang, Shengyong

AU - Qiu, Yan

AU - You, Bo

AU - Chen, Weijun

PY - 2008

Y1 - 2008

N2 - Background. In the absence of effective drugs, controlling SARS relies on the rapid identification of cases and appropriate management of the close contacts, or effective vaccines for SARS. Therefore, developing specific and sensitive laboratory tests for SARS as well as effective vaccines are necessary for national authorities. Results. Genes encoding truncated nucleocapsid (N) and spike (S) proteins of SARSCoV were cloned into the expression vector pQE30 and fusionally expressed in Escherichia coli M15. The fusion protein was analyzed for reactivity with SARS patients' sera and with anti-sera against the two human coronaviruses HCoV 229E and HCoV OC43 by ELISA, IFA and immunoblot assays. Furthermore, to evaluate the antigen-specific humoral antibody and T-cell responses in mice, the fusion protein was injected into 6-week-old BALB/c mice and a neutralization test as well as a T-cell analysis was performed. To evaluate the antiviral efficacy of immunization, BALB/c mice were challenged intranasally with SARSCoV at day 33 post injection and viral loads were determined by fluorescent quantitative RT-PCR. Serological results showed that the diagnostic sensitivity and specificity of the truncated S-N fusion protein derived the SARS virus were > 99% (457/460) and 100.00% (650/650), respectively. Furthermore there was no cross-reactivity with other two human coronaviruses. High titers of antibodies to SRASCoV appeared in the immunized mice and the neutralization test showed that antibodies to the fusion protein could inhibit SARSCoV. The T cell proliferation showed that the fusion protein could induce an antigen-specific T-cell response. Fluorescent quantitative RT-PCR showed that BALB/c mice challenged intranasally with SARSCoV at day 33 post injection were completely protected from virus replication. Conclusion. The truncated S-N fusion protein is a suitable immunodiagnostic antigen and vaccine candidate.

AB - Background. In the absence of effective drugs, controlling SARS relies on the rapid identification of cases and appropriate management of the close contacts, or effective vaccines for SARS. Therefore, developing specific and sensitive laboratory tests for SARS as well as effective vaccines are necessary for national authorities. Results. Genes encoding truncated nucleocapsid (N) and spike (S) proteins of SARSCoV were cloned into the expression vector pQE30 and fusionally expressed in Escherichia coli M15. The fusion protein was analyzed for reactivity with SARS patients' sera and with anti-sera against the two human coronaviruses HCoV 229E and HCoV OC43 by ELISA, IFA and immunoblot assays. Furthermore, to evaluate the antigen-specific humoral antibody and T-cell responses in mice, the fusion protein was injected into 6-week-old BALB/c mice and a neutralization test as well as a T-cell analysis was performed. To evaluate the antiviral efficacy of immunization, BALB/c mice were challenged intranasally with SARSCoV at day 33 post injection and viral loads were determined by fluorescent quantitative RT-PCR. Serological results showed that the diagnostic sensitivity and specificity of the truncated S-N fusion protein derived the SARS virus were > 99% (457/460) and 100.00% (650/650), respectively. Furthermore there was no cross-reactivity with other two human coronaviruses. High titers of antibodies to SRASCoV appeared in the immunized mice and the neutralization test showed that antibodies to the fusion protein could inhibit SARSCoV. The T cell proliferation showed that the fusion protein could induce an antigen-specific T-cell response. Fluorescent quantitative RT-PCR showed that BALB/c mice challenged intranasally with SARSCoV at day 33 post injection were completely protected from virus replication. Conclusion. The truncated S-N fusion protein is a suitable immunodiagnostic antigen and vaccine candidate.

UR - http://www.scopus.com/inward/record.url?scp=58149265725&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=58149265725&partnerID=8YFLogxK

U2 - 10.1186/1471-2180-8-207

DO - 10.1186/1471-2180-8-207

M3 - Article

VL - 8

JO - BMC Microbiology

JF - BMC Microbiology

SN - 1471-2180

M1 - 207

ER -