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Journal of Virology, June 2004, p. 5612-5618, Vol. 78, No. 11
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.11.5612-5618.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

T-Cell Epitopes in Severe Acute Respiratory Syndrome (SARS) Coronavirus Spike Protein Elicit a Specific T-Cell Immune Response in Patients Who Recover from SARS

Yue-Dan Wang,1,2,{dagger} Wan-Yee Fion Sin,2,{dagger} Guo-Bing Xu,3,{dagger} Huang-Hua Yang,4 Tin-yau Wong,5 Xue-Wen Pang,1 Xiao-Yan He,1 Hua-Gang Zhang,1 Joice Na Lee Ng,4 Chak-Sum Samuel Cheng,2 Jing Yu,2 Li Meng,2 Rui-Feng Yang,3 Sik-To Lai,5 Zhi-Hong Guo,4 Yong Xie,2* and Wei-Feng Chen1*

Department of Immunology, Peking University Health Science Centre,1 First Hospital, Peking University, Beijing,3 Department of Biology,2 Department of Chemistry, Hong Kong University of Science and Technology, Clear Water Bay, Kowloon,4 Princess Margaret Hospital, Hong Kong, People's Republic of China5

Received 3 November 2003/ Accepted 15 January 2004

The immunogenicity of HLA-A2-restricted T-cell epitopes in the S protein of the Severe acute respiratory syndrome coronavirus (SARS-CoV) and of human coronavirus strain 229e (HCoV-229e) was analyzed for the elicitation of a T-cell immune response in donors who had fully recovered from SARS-CoV infection. We employed online database analysis to compare the differences in the amino acid sequences of the homologous T epitopes of HCoV-229e and SARS-CoV. The identified T-cell epitope peptides were synthesized, and their binding affinities for HLA-A2 were validated and compared in the T2 cell system. The immunogenicity of all these peptides was assessed by using T cells obtained from donors who had fully recovered from SARS-CoV infection and from healthy donors with no history of SARS-CoV infection. HLA-A2 typing by indirect immunofluorescent antibody staining showed that 51.6% of SARS-CoV-infected patients were HLA-A2 positive. Online database analysis and the T2 cell binding test disclosed that the number of HLA-A2-restricted immunogenic epitopes of the S protein of SARS-CoV was decreased or even lost in comparison with the homologous sequences of the S protein of HCoV-229e. Among the peptides used in the study, the affinity of peptides from HCoV-229e (H77 and H881) and peptides from SARS-CoV (S978 and S1203) for binding to HLA-A2 was higher than that of other sequences. The gamma interferon (IFN-{gamma}) release Elispot assay revealed that only SARS-CoV-specific peptides S1203 and S978 induced a high frequency of IFN-{gamma}-secreting T-cell response in HLA-A2+ donors who had fully recovered from SARS-CoV infection; such a T-cell epitope-specific response was not observed in HLA-A2+ healthy donors or in HLA-A2– donors who had been infected with SARS-CoV after full recovery. Thus, T-cell epitopes S1203 and S978 are immunogenic and elicit an overt specific T-cell response in HLA-A2+ SARS-CoV-infected patients.

* Corresponding author. Mailing address for Wei-Feng Chen: Department of Immunology, Peking University Health Science Centre, 38, Xueyuanlu, Beijing, 100083, People's Republic of China. Phone: (86) 10-82802593. Fax: (86) 10-82802593. E-mail: wfchen{at}public.bta.net.cn. Mailing address for Yong Xie: Department of Biology, Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong, People's Republic of China. Phone: (852)2358-7340. Fax: (852)2358-1559. E-mail: Boyxie{at}ust.hk.

{dagger} Y.-D.W., W.-Y.F.S., and G.-B.X. contributed equally to this study.

Journal of Virology, June 2004, p. 5612-5618, Vol. 78, No. 11
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.11.5612-5618.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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