title: Induction of immune response against NY-ESO-1 by CHP-NY-ESO-1 vaccination and immune regulation in a melanoma patient.
abstract: BACKGROUND
  NY-ESO-1 is a cancer/testis antigen highly immunogenic in cancer patients. Cholesterol-bearing hydrophobized pullulan (CHP) is a nanoparticle-forming antigen-delivery vehicle and CHP complexed with NY-ESO-1 protein (CHP-NY-ESO-1) efficiently activates CD4 and CD8 T cells in vitro.
  
  
  OBJECTIVE
  In this study we report on a 50-year-old male melanoma patient with multiple skin and organ metastases (T4N3M1c) who was vaccinated with CHP-NY-ESO-1 at biweekly intervals and who had an unusual disease course. We characterized in this patient humoral and cellular immune responses, immune regulatory cells, and cytokine profiles in the peripheral blood and at local tumor sites.
  
  
  RESULTS
  Ten days after the second CHP-NY-ESO-1 vaccination (day 25), blisters appeared on the skin at the metastatic lesions associated with inflammatory changes. A skin biopsy showed the presence of many NY-ESO-1-expressing apoptotic melanoma cells as determined by a terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling (TUNEL) test. However, the tumors continued to grow, and the patient died of pulmonary failure due to multiple metastases on day 48. Serum antibody responses were detected after the second CHP-NY-ESO-1 vaccination and antibody titer increased with subsequent vaccinations. Th1 dependent IgG1 was the predominant immunoglobulin subtype. Both, NY-ESO-1-specific CD4 and CD8 T cell responses were detected in PBMC by IFN-gamma secretion assays. After CHP-NY-ESO-1 vaccination a slight decrease in CD4(+)CD25(+)Foxp3(+) Tregs was observed in PBMC but significantly increased numbers of CD4(+)CD25(+)Foxp3(+) Tregs and CD68(+) immunoregulatory macrophages were detected at the local tumor sites. CD4(+)CD25(+)Foxp3(+) Tregs were also increased in the blister fluid. Cytokines in the serum suggested a polarization towards a Th1 pattern in the PBMC and those in the blister fluid suggested a Th2-type response at the tumor site.
  
  
  CONCLUSIONS
  Our observations indicate induction of specific humoral and cellular immune responses against NY-ESO-1 after CHP-NY-ESO-1 vaccination in a melanoma patient. The concomitant appearance of regulatory T cells and of immune regulatory macrophages and cytokines at the local tumor sites in this patient may explain immune escape.
authors.affiliation: Department of Dermatology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, Japan. tsujikaz@cc.okayama-u.ac.jp
authors.forename: Kazuhide
authors.initials: K
authors.lastname: Tsuji
authors.forename: Toshitada
authors.initials: T
authors.lastname: Hamada
authors.forename: Akiko
authors.initials: A
authors.lastname: Uenaka
authors.forename: Hisashi
authors.initials: H
authors.lastname: Wada
authors.forename: Eiichi
authors.initials: E
authors.lastname: Sato
authors.forename: Midori
authors.initials: M
authors.lastname: Isobe
authors.forename: Kenji
authors.initials: K
authors.lastname: Asagoe
authors.forename: Osamu
authors.initials: O
authors.lastname: Yamasaki
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Shiku
authors.forename: Gerd
authors.initials: G
authors.lastname: Ritter
authors.forename: Roger
authors.initials: R
authors.lastname: Murphy
authors.forename: Eric W
authors.initials: EW
authors.lastname: Hoffman
authors.forename: Lloyd J
authors.initials: LJ
authors.lastname: Old
authors.forename: Eiichi
authors.initials: E
authors.lastname: Nakayama
authors.forename: Keiji
authors.initials: K
authors.lastname: Iwatsuki
mesh_terms: D000912:Antibodies, Neoplasm / Q000097:blood; D000951:Antigens, Neoplasm / Q000276:immunology / Q000627:therapeutic use*; D017209:Apoptosis; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D019496:Cancer Vaccines / Q000627:therapeutic use*; D016207:Cytokines / Q000032:analysis / Q000276:immunology; D005455:Fluorescent Antibody Technique; D005936:Glucans / Q000276:immunology / Q000627:therapeutic use*; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood; D020287:In Situ Nick-End Labeling; D008264:Macrophages / Q000276:immunology; D008297:Male; D008545:Melanoma / Q000276:immunology / Q000473:pathology / Q000628:therapy*; D008565:Membrane Proteins / Q000276:immunology / Q000627:therapeutic use*; D008875:Middle Aged; D012878:Skin Neoplasms / Q000276:immunology / Q000473:pathology / Q000628:therapy*; D016176:T-Lymphocyte Subsets / Q000276:immunology
chemical_list: D000912:Antibodies, Neoplasm; D000951:Antigens, Neoplasm; C104948:CTAG1B protein, human; D019496:Cancer Vaccines; D016207:Cytokines; D005936:Glucans; D007074:Immunoglobulin G; D008565:Membrane Proteins; C009109:pullulan