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Identification of cytotoxic T lymphocyte epitopes in dengue virus serotype 1
Author(s) -
Duan Zhiliang,
Guo Jianglong,
Huang Xi,
Liu Huifang,
Chen Xinyu,
Jiang Minghua,
Wen Jinsheng
Publication year - 2015
Publication title -
journal of medical virology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.782
H-Index - 121
eISSN - 1096-9071
pISSN - 0146-6615
DOI - 10.1002/jmv.24167
Subject(s) - epitope , dengue virus , virology , biology , elispot , cytotoxic t cell , immunogenicity , peptide vaccine , microbiology and biotechnology , dengue vaccine , cd8 , dengue fever , antigen , immunology , biochemistry , in vitro
Dengue virus (DENV) has a serious and growing impact on global health and the exact role of DENV‐specific CD8 + T‐cells in DENV infection is still uncertain. In the present study, SYFPEITHI algorithm was used to screen the amino acid sequence of Dengue virus serotype 1 (DENV‐1) for potential epitopes, and seven putative HLA‐A*1101‐restricted and five putative HLA‐A*2402‐restricted epitopes conserved in hundreds of DENV‐1 strains were synthesized. The binding affinity of these epitope candidates to corresponding HLA molecules was evaluated using competitive peptide‐binding assay. The immunogenicity and specificity of peptides were further tested in HLA‐A*1101 transgenic mice, HLA‐A*2402 transgenic mice and peripheral blood mononuclear cells (PBMCs) of patients infected with DENV‐1. Percentage inhibition (PI) values calculated in competitive peptide‐binding assay showed that six peptides (E 39‐47 PTLDIELLK, NS5 505‐513 GVEGEGLHK, NS2b 15‐23 SILLSSLLK, NS5 561‐569 ALLATSIFK, NS3 99‐107 AVEPGKNPK, and NS4b 159‐167 VVYDAKFEK) could bind to HLA‐A*1101 molecule with high affinity and five peptides (NS3 472‐480 QYIYMGQPL, NS4a 40‐48 AYRHAMEEL, NS5 880‐888 DYMTSMKRF, NS3 548‐556 SYKVASEGF, and NS3 22‐30 IYRILQRGL) have a high affinity for HLA‐A*2402 molecule. Enzyme‐linked immunospot (ELISPOT) results indicated that these high‐affinity peptides were recognized by splenocytes of DENV‐1‐infected transgenic mice and high‐affinity peptide‐immunized transgenic mice displayed high levels of peptide‐specific IFN‐γ‐secreting cells. In addition, both peptide‐pulsed splenocytes and DENV‐1‐infected splenic monocytes were efficiently killed by these peptide‐specific cytotoxic T lymphocytes. Finally, except NS2b 15‐23 , 10 high‐affinity peptides were recognized by PBMCs of patients infected with DENV‐1. These identified epitopes would contribute to the understanding of the function of DENV‐specific CD8 + T‐cells. J. Med. Virol. 87:1077–1089, 2015 . © 2015 Wiley Periodicals, Inc.