title: Identification of conserved and HLA-A*2402-restricted epitopes in Dengue virus serotype 2.
abstract: In this study, we set out to identify dengue virus serotype 2 (DENV-2)-specific HLA-A*2402-restricted epitopes and determine the characteristics of T cells generated to these epitopes. We screened the full-length amino-acid sequence of DENV-2 to find potential epitopes using the SYFPEITHI algorithm. Twelve putative HLA-A*2402-binding peptides conserved in hundreds of DENV-2 strains were synthesized, and the HLA restriction of peptides was tested in HLA-A*2402 transgenic mice. Nine peptides (NS4b(228-237), NS2a(73-81), E(298-306), M(141-149), NS4a(96-105), NS4b(159-168), NS5(475-484), NS1(162-171), and NS5(611-620)) induced high levels of peptide-specific IFN-γ-secreting cells in HLA-A*2402 transgenic mice. Apart from IFN-γ, NS4b(228-237-), NS2a(73-81-) and E(298-306)-specific CD8(+) cells produced TNF-α and IL-6 simultaneously, whereas M(141-149-) and NS5(475-484-) CD8(+) cells produced only IL-6. Moreover, splenic mononuclear cells (SMCs) efficiently recognized and killed peptide-pulsed splenocytes. Furthermore, each of nine peptides could be recognized by splenocytes from DENV-2-infected HLA-A*2402 transgenic mice. The SMCs from HLA-A*2402 transgenic mice immunized with nine immunogenic peptides efficiently killed DENV-2-infected splenic monocytes. The present identified epitopes have the potential to be new diagnostic tools for characterization of T-cell immunity in DENV infection and may serve as part of a universal epitope-based vaccine.
authors.affiliation: Department of Microbiology and Immunology, Wenzhou Medical University, Wenzhou, China; Department of Clinical Laboratory, The Second Affiliated Hospital & Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, China; Institute of Arboviruses, School of Basic Medical Sciences, Wenzhou Medical University, Wenzhou, China.
authors.forename: Zhi-Liang
authors.initials: ZL
authors.lastname: Duan
authors.affiliation: Department of Microbiology and Immunology, Wenzhou Medical University, Wenzhou, China.
authors.forename: Hui-Fang
authors.initials: HF
authors.lastname: Liu
authors.affiliation: Department of Immunology, Institute of Tuberculosis Control, Institute of Human Virology, Sun Yat-sen University, Zhongshan School of Medicine, Guangzhou, China; Key Laboratory of Tropical Diseases Control (Sun Yat-sen University), Ministry of Education, Guangzhou, China.
authors.forename: Xi
authors.initials: X
authors.lastname: Huang
authors.affiliation: Department of Microbiology and Immunology, Wenzhou Medical University, Wenzhou, China.
authors.forename: Si-Na
authors.initials: SN
authors.lastname: Wang
authors.affiliation: Department of Microbiology and Immunology, Wenzhou Medical University, Wenzhou, China.
authors.forename: Jin-Lin
authors.initials: JL
authors.lastname: Yang
authors.affiliation: Department of Clinical Laboratory, The Second Affiliated Hospital & Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, China.
authors.forename: Xin-Yu
authors.initials: XY
authors.lastname: Chen
authors.affiliation: Department of Liver, The Secondary Hospital of Ningbo, Ningbo, China.
authors.forename: De-Zhou
authors.initials: DZ
authors.lastname: Li
authors.affiliation: Department of Microbiology and Immunology, Wenzhou Medical University, Wenzhou, China.
authors.forename: Xiao-Zhi
authors.initials: XZ
authors.lastname: Zhong
authors.affiliation: Department of Microbiology and Immunology, Wenzhou Medical University, Wenzhou, China.
authors.forename: Bo-Kun
authors.initials: BK
authors.lastname: Chen
authors.affiliation: Department of Microbiology and Immunology, Wenzhou Medical University, Wenzhou, China; Institute of Arboviruses, School of Basic Medical Sciences, Wenzhou Medical University, Wenzhou, China. Electronic address: wjs78@wmu.edu.cn.
authors.forename: Jin-Sheng
authors.initials: JS
authors.lastname: Wen
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D016207:Cytokines / Q000378:metabolism; D003715:Dengue / Q000276:immunology* / Q000378:metabolism; D003716:Dengue Virus / Q000145:classification / Q000276:immunology*; D004195:Disease Models, Animal; D018604:Epitope Mapping; D000939:Epitopes / Q000737:chemistry / Q000276:immunology*; D018984:Epitopes, T-Lymphocyte / Q000737:chemistry / Q000276:immunology; D005260:Female; D015234:HLA-A Antigens / Q000276:immunology*; D006801:Humans; D007114:Immunization; D016130:Immunophenotyping; D008822:Mice, Transgenic; D010455:Peptides / Q000737:chemistry / Q000276:immunology; D065288:Serogroup; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology / Q000378:metabolism
chemical_list: D016207:Cytokines; D000939:Epitopes; D018984:Epitopes, T-Lymphocyte; D015234:HLA-A Antigens; D010455:Peptides
keywords: Dengue virus; Epitope; HLA-A*2402