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Immune response in Balb/c mice induced by recombinant spike protein of severe acute respiratory syndrome coronavirus

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Chinese Medical Journal 2005年第10期118卷 857-860页

作者： QINLi WUShao-ting WANGXi-ming YUANShi-shan HUANGDa-na LEIMing-jun PANHui-rong LINQi-ping GAOShi-tong ZHANGRen-li QUShen DepartmentofBiochemistryandMolecularBiology TongjiMedicalCollegeHuazhongUniversityofScienceandTechnologyWuhan430030China ShenzhenCenterforDiseaseControlandPrevention Shenzhen518020China

Severe acute respiratory syndrome coronavirus (SARS-CoV) is a highly aggressive pathogen that caused SARS in 2003 and 2004.^(1-4) Spike protein (S) on the surface of virus particles mediates the attachment of the virus to cell surface receptors and induces the fusion of viral and cellular membranes. According to the epitope analysis and structure of S protein, we used two fragments of S protein S1 (108-488aa) and S2 (723-938aa) expressed in Escherichia coli and immunized Balb/c mice to investigate the immune response to the recombinant proteins in mice.~5

关键词： severe acute respiratory syndrome coronavirus spike protein igg isotype T lymphocyte subset cytokine

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The immune response in Balb/c mice induced by recombinant spike protein of SARS coronavirus

The immune response in Balb/c mice induced by recombinant sp...

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全国寄生虫学与热带医学学术研讨会

作者：秦莉吴少庭王西明袁仕善黄达娜雷明军潘辉榕林绮萍高世同张仁利屈伸 Department of Biochemistry and Molecular Biology Tongji Medical CollegeHuazhong University of Science and Technology Shenzhen Center for Disease Control and Prevention Veterinarian College South China University of Agriculture

Background The severe acute respiratory syndrome(SARS)-associated coronavirus,namely SARS-CoV, causes SARS for which there are no vaccines or effective therapies currently available.A surface projection glycoprotein of the virus particle,spike protein plays an important role in SARS-CoV infection and it is a main protective antigen *** this case spike protein may be a good candidate vaccine molecular to SARS. Method Two SARS coronavirus spike gene fragments S1(322-1464bp) and S2(2170-2814bp) were amplified by RT-PCR from the SARS coronavirus culture supernatant and cloned into pET-23a vector,and expressed in Escherichia coli BL21(DE3)(pLyS) *** proteins were purified by Ni2+ immobilized metal ion affinity chromatography and injected into Balb/c mice mixed with complete Freund adjuvant three times for two weeks *** substantial immune responses were measured by serum antibody titer and cytokine concentration using enzyme-linked immunosorben assay(ELISA) and spleen T cells subset using flow cytometry. Result The two plasmids were constructed successfully.25KD and 45KD recombinant proteins were purified from the periplasmic fraction and from insoluble fraction of the inclusion bodies and recognized by the serum of convalescent SARS *** titer antibody was detected in the serum of immunized Balb/c *** mice injected with S1 protein produced high titer igg1,igg2a,igg2b and igg3 while the mice injected with S2 protein produced high titer igg1,igg2a,but lower titer igg2b and *** cytokine concentration did not changed distinctively in immunized *** the percentage of spleen CD8+ T cells also increased in immunized mice. Conclusion The two recombinant proteins of SARS coronavirus spike protein induced hormonal and cellular immune response in Balb/c mice.

关键词： severe acute respiratory syndrome coronavirus spike protein igg isotype T lymphocyte subset cytokine

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