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Identification of an epitope of SARS-coronavirus nucleocapsid protein

Cell Research 2003年第3期13卷 141-145页

作者： YING LIN,XU SHEN,RUI FU YANG,YI XUE LI,YONG YONG JI,YOU Yu HE,MU DE SHI,WEI LU,TIE LIU SHI,JIN WANG,HONG XIA WANG,HUA LIANG JIANG,JIAN HUA SHEN,You HUA XIE,YUAN WANG,GANG PEI,BEI FEN SHEN,JIA RUI WU,BING SUNInstitute of Biochemistry and Cell Biology,Institute of Materia Medico,,Bioinformation Center,Shanghai Institutes for Biological Sciences,Chinese Academy of Sciences. Shanghai 200031,ChinaInstitute of Microbiology and Epidemiology,Institute of Basic Medical Sciences,Academy of Military Medical,Sc.imce.fi,Beijing 100071,China Institute of Biochemistry and Cell Biology Chinese Academy of Sciences Shanghai 200031 China.

The nucleocapsid(N)protein of severe acute respiratory syndrome-coronavirus(SARS-CoV)is a majorvirion structural *** this study,two epitopes(N1 and N2)of the N protein of SARS-CoV werepredicted by bioinformatics *** immnnization with two peptides,the peptides-specific antibodieswere isolated from the immunized *** further experiments demonstrated that N1 peptide-inducedpolyclonal antibodies had a high affinity to bind to *** expressed Nprotein of ***,itwas confirmed that N1 peptide-specific IgG antibodies were detectable in the sera of severe acute respiratorysyndrome(SARS)*** results indicated that an epitope of the N protein has been identified andN protein specific Abs were produced by peptide immunization,which will be useful for the study of SARS-CoV.

关键词： severe acute respiratory syndrome-coronavirus necleocapsid protein epitope polyclonal antibody antiserum.

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Immune Responses Against SARS-coronavirus Nucleocapsid Protein Induced by DNA Vaccine

Immune Responses Against SARS-Coronavirus Nucleocapsid Prote...

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2005全国第二届核酸疫苗研讨会

作者： ZHAO Ping PAN Wei KE Jin-Shan QI Zhong-Tian (Department of Microbiology, Second Military Medical University, Shanghai 200433, China)

The nucleocapsid (N) protein of SARS-coronavirus (SARS-CoV) is the key protein for the formation of the helical nucleosapsid during virion assembly. This protein is believed to be more conserved than other proteins of the virus, such as spike and membrane glycoprotein. In this study, the N protein of SARS-CoV was expressed in *** DH5 and identified with pooled sera from patients in the convalescence phase of SARS. A plasmid pCI-N, encoding the full-length N gene of SARS-CoV, was constructed. Expression of the N protein was observed in COS1 cells following transfection with pCI-N. The immune responses induced by intramuscular immunization with pCI-N was evaluated in a murine model. Serum anti-N immunoglobulins and splenocytes proliferative responses against N protein were observed in immunized BALB/c mice. The major immunoglobulin G sublass recognizing N protein was immunoglobulin G2a, and stimulated splenocytes secreted high levels of gamma interferon and IL-2 in response to N protein. More importantly, the immunized mice produced strong delayed-type hypersensitivity (DTH) and CD8+ CTL responses to N protein. The study shows that N protein of SARS-CoV not only is an important B cell immunogen, but also can elicit broad-based cellular immune responses. The results indicate that the N protein may be of potential value in vaccine development for specific prophylaxis and treatment against SARS.

关键词： severe acute respiratory syndrome-coronavirus nucleocapsid protein immune response cell-mediated immunity DNA vaccine

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