title: The spontaneous CD8+ T-cell response to HLA-A2-restricted NY-ESO-1b peptide in hepatocellular carcinoma patients.
abstract: OBJECTIVE
  Hepatocellular carcinoma (HCC) can express various cancer-testis antigens including NY-ESO-1, members of the SSX family, members of the MAGE family, SCP-1, and CTP11. Immunotherapy directed against these antigens is a potential alternative treatment for HCC. To date, it remains unclear whether HCC patients have spontaneous immune responses to these tumor antigens. The objectives of this study were to measure immune responses to NY-ESO-1, a promising cancer vaccine candidate, in HCC patients using the HLA-A2-restricted NY-ESO-1b peptide (p157-165) to measure cellular responses and whole protein to measure antibody responses.
  
  
  METHODS
  In HLA-A2(+) patients with NY-ESO-1(+) HCC, we analyzed T-cell antigen-dependent interferon (IFN)-gamma and/or Granzyme B release by enzyme-linked immunospot (ELISPOT) assay and IFN-gamma-producing intracellular cytokine flow cytometry (CytoSpot). As an assay independent of T-cell function, we performed tetramer staining. Antibodies to whole NY-ESO-1 were assayed by enzyme-linked immunosorbent assay.
  
  
  RESULTS
  The frequency of specific CD8(+) T-cell responses to NY-ESO-1b in 28 NY-ESO-1 mRNA(+)HLA-A2(+) HCC patients was 35.7% (10 of 28). The average magnitude of effector CD8(+) T cells was 0.3% (89 +/- 59 per 2.5 x 10(4) CD8(+) cells) and 1.2% as measured by IFN-gamma release ELISPOT and CytoSpot assays, respectively. These in vitro induced NY-ESO-1b-specific CD8(+) T cells can also recognize HepG2 cells transfected with pcDNA3.1-NY-ESO-1 in both IFN-gamma and Granzyme B ELISPOT assays. Frequencies of NY-ESO-1b-specific T cells in several patients were confirmed by tetramer staining. Nonfunctional tetramer(+)CD8(+) T cells were also present. The CD8(+) T-cell response was apparently increased in patients with late-stage HCC. A discordance between antibody and CD8(+) T-cell responses in HCC patients was observed.
  
  
  CONCLUSIONS
  The elevated frequency of specific CD8(+) T-cell responses to NY-ESO-1b in NY-ESO-1 mRNA(+)HLA-A2(+) HCC patients suggests that NY-ESO-1 is appropriate for use in the immunotherapy of HCC patients.
authors.affiliation: Department of Immunology, School of Basic Medical Sciences, Peking University Health Science Center, Beijing, People's Republic of China.
authors.forename: Xiao-Ying
authors.initials: XY
authors.lastname: Shang
authors.forename: Hong-Song
authors.initials: HS
authors.lastname: Chen
authors.forename: Hua-Gang
authors.initials: HG
authors.lastname: Zhang
authors.forename: Xue-Wen
authors.initials: XW
authors.lastname: Pang
authors.forename: Huan
authors.initials: H
authors.lastname: Qiao
authors.forename: Ji-Run
authors.initials: JR
authors.lastname: Peng
authors.forename: Li-Ling
authors.initials: LL
authors.lastname: Qin
authors.forename: Ran
authors.initials: R
authors.lastname: Fei
authors.forename: Ming-Hui
authors.initials: MH
authors.lastname: Mei
authors.forename: Xi-Sheng
authors.initials: XS
authors.lastname: Leng
authors.forename: Sacha
authors.initials: S
authors.lastname: Gnjatic
authors.forename: Gerd
authors.initials: G
authors.lastname: Ritter
authors.forename: Andrew J G
authors.initials: AJ
authors.lastname: Simpson
authors.forename: Lloyd J
authors.initials: LJ
authors.lastname: Old
authors.forename: Wei-Feng
authors.initials: WF
authors.lastname: Chen
mesh_terms: D000328:Adult; D000368:Aged; D000917:Antibody Formation; D000951:Antigens, Neoplasm / Q000494:pharmacology*; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D019496:Cancer Vaccines / Q000276:immunology*; D006528:Carcinoma, Hepatocellular / Q000276:immunology*; D005260:Female; D005434:Flow Cytometry; D053804:Granzymes; D015789:HLA-A2 Antigen / Q000276:immunology; D006801:Humans; D007118:Immunoassay; D007167:Immunotherapy / Q000379:methods; D007371:Interferon-gamma / Q000494:pharmacology; D008113:Liver Neoplasms / Q000276:immunology*; D008297:Male; D008565:Membrane Proteins / Q000494:pharmacology*; D008875:Middle Aged; D010446:Peptide Fragments; D012697:Serine Endopeptidases / Q000494:pharmacology
chemical_list: D000951:Antigens, Neoplasm; C104948:CTAG1B protein, human; D019496:Cancer Vaccines; D015789:HLA-A2 Antigen; D008565:Membrane Proteins; D010446:Peptide Fragments; D007371:Interferon-gamma; C509213:GZMB protein, human; D053804:Granzymes; D012697:Serine Endopeptidases