title: Identification of HLA-A*2402-restricted HCMV immediate early-1 (IE-1) epitopes as targets for CD8+ HCMV-specific cytotoxic T lymphocytes.
abstract: BACKGROUND
  To identify novel HLA-A*2402-restricted human cytomegalovirus (HCMV) immediate early-1 (IE-1) epitopes for adoptive immunotherapy, we explored 120 overlapping 15-amino acid spanning IE-1.
  
  
  METHODS
  These peptides were screened by measuring the frequency of polyclonal CD8+ T cells producing intracellular interferon-gamma (IFN-gamma) using flow cytometry and the epitopes were validated with a HCMV-infected target Cr release cytotoxicity assay.
  
  
  RESULTS
  Initial screening was performed with 12 mini-pools of 10 consecutive peptides made from 120 overlapping peptides15-amino acids in length that spanned IE-1. When peripheral blood mononuclear cells (PBMCs) from HLA-A*2402 HCMV-seropositive donors were sensitized with each of the 12 mini-pools, mini-pools 1 and 2 induced the highest frequency of CD8+ cytotoxic T lymphocytes (CTLs) producing IFN-gamma. When PBMCs were stimulated with each of the twenty peptides belonging to mini-pools 1 and 2, peptides IE-11-15MESSAKRKMDPDNPD and IE-15-19AKRKMDPDNPDEGPS induced the greatest quantities of IFN-gamma production and cytotoxicity of HLA-matched HCMV-infected fibroblasts. To determine the exact HLA-A*2402-restricted epitopes within the two IE-1 proteins, we synthesized a total of twenty-one overlapping 9- or 10 amino acid peptides spanning IE-11-15 and IE-15-19. Peptide IE-13-12SSAKRKMDPD induced the greatest quantities of IFN-gamma production and target cell killing by CD8+ CTLs.
  
  
  CONCLUSIONS
  HCMV IE-13-12SSAKRKMDPD is a HLA-A*2402-restricted HCMV IE-1 epitope that can serve as a common target for CD8+ HCMV-specific CTLs.
authors.affiliation: Department of Transfusion Medicine, Warren G, Magnuson Clinical Center, National Institutes of Health, Bethesda, MD, USA. jlim@yumc.yonsei.ac.kr
authors.forename: Jong-Baeck
authors.initials: JB
authors.lastname: Lim
authors.forename: Hyun Ok
authors.initials: HO
authors.lastname: Kim
authors.forename: Seok Hoon
authors.initials: SH
authors.lastname: Jeong
authors.forename: Joo Eun
authors.initials: JE
authors.lastname: Ha
authors.forename: Sunphil
authors.initials: S
authors.lastname: Jang
authors.forename: Sang-Guk
authors.initials: SG
authors.lastname: Lee
authors.forename: Kyungwon
authors.initials: K
authors.lastname: Lee
authors.forename: David
authors.initials: D
authors.lastname: Stroncek
mesh_terms: D000595:Amino Acid Sequence; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology* / Q000821:virology; D002460:Cell Line; D003587:Cytomegalovirus / Q000276:immunology*; D003586:Cytomegalovirus Infections / Q000276:immunology / Q000628:therapy; D000939:Epitopes / Q000235:genetics / Q000276:immunology*; D005347:Fibroblasts / Q000166:cytology / Q000276:immunology; D015234:HLA-A Antigens / Q000276:immunology*; D059871:HLA-A24 Antigen; D006801:Humans; D017874:Immediate-Early Proteins / Q000235:genetics / Q000276:immunology*; D007371:Interferon-gamma / Q000276:immunology; D008969:Molecular Sequence Data; D010455:Peptides / Q000235:genetics / Q000276:immunology; D016176:T-Lymphocyte Subsets / Q000276:immunology* / Q000821:virology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology* / Q000821:virology; D014764:Viral Proteins / Q000235:genetics / Q000276:immunology
chemical_list: D000939:Epitopes; D015234:HLA-A Antigens; C438506:HLA-A*24:02 antigen; D059871:HLA-A24 Antigen; D017874:Immediate-Early Proteins; D010455:Peptides; D014764:Viral Proteins; D007371:Interferon-gamma
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