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Journal of Virology, June 2007, p. 6079-6088, Vol. 81, No. 11
0022-538X/07/$08.00+0     doi:10.1128/JVI.02568-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Identification and Characterization of Dominant Helper T-Cell Epitopes in the Nucleocapsid Protein of Severe Acute Respiratory Syndrome Coronavirus{triangledown}

Jincun Zhao,{dagger} Qianrong Huang,{dagger} Wei Wang, Yan Zhang, Ping Lv, and Xiao-Ming Gao*

Department of Immunology, Peking University Health Science Center, Peking University, 100083 Beijing, China

Received 21 November 2006/ Accepted 16 March 2007

By using a series of overlapping synthetic peptides covering 98% of the amino acid sequence of the nucleocapsid protein (NP) of severe acute respiratory syndrome coronavirus (SARS-CoV), four helper T-cell (Th) epitopes (NP11, residues 11 to 25; NP51, residues 51 to 65; NP61, residues 61 to 75; and NP111, residues 111 to 125) in C57BL mice (H-2b), four (NP21, residues 21 to 35; NP91, residues 91 to 105; NP331, residues 331 to 345; and NP351, residues 351 to 365) in C3H mice (H-2k), and two (NP81, residues 81 to 95; and NP351, residues 351 to 365) in BALB/c mice (H-2d) have been identified. All of these peptides were able to stimulate the proliferation of NP-specific T-cell lines or freshly isolated lymph node cells from mice immunized with recombinant NP. Immunization of mice with synthetic peptides containing appropriate Th epitopes elicited strong cellular immunity in vivo, as evidenced by delayed-type hypersensitivity. Priming with the helper peptides (e.g., NP111 and NP351) significantly accelerated the immune response induced by recombinant NP, as determined by the production of NP-specific antibodies. When fused with a conserved neutralizing epitope (SP1143-1157) from the spike protein of SARS-CoV, NP111 and NP351 assisted in the production of high-titer neutralizing antibodies in vivo. These data provide useful insights regarding immunity against SARS-CoV and have the potential to help guide the design of peptide-based vaccines.

* Corresponding author. Mailing address: Department of Immunology, Peking University Health Science Center, 38 Xueyuan Road, Beijing 100083, China. Phone and fax: 86 10 82801156. E-mail: xmgao{at}bjmu.edu.cn

{triangledown} Published ahead of print on 28 March 2007.

{dagger} These authors contributed equally to this work.

Journal of Virology, June 2007, p. 6079-6088, Vol. 81, No. 11
0022-538X/07/$08.00+0     doi:10.1128/JVI.02568-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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