title: Expansion of melanoma-specific T cells from lymph nodes of patients in stage III: implications for adoptive immunotherapy in treating cancer.
abstract: BACKGROUND
  Adoptive immunotherapy for patients with metastatic melanoma has yielded encouraging results. However, methods of expanding melanoma-specific T cells from stage III are limited. The objective of this study was to determine whether melanoma-specific T cells could be generated from the melanoma-draining lymph nodes (MDLNs) of patients in stage III.
  
  
  METHODS
  Patients in stage III who were undergoing completion lymphadenectomy were enrolled into a protocol approved by the institutional review board. MDLN cells were tested for ability to undergo cryopreservation, expand ex vivo in IL-2 or IL-2 and IL-7, and mediate melanoma-specific antitumor responses in vitro.
  
  
  RESULTS
  Cryopreservation produced no significant differences from fresh cultures in terms of cell growth and cellular phenotype. IL-2 and IL-2/IL-7 cultures resulted in similar growth rates, and functional studies revealed the presence of T cells that secreted interferon gamma in response to melanoma antigen peptides. Both IL-2- and IL-2/IL-7-cultured MDLN cells mediated significant apoptosis of human melanoma cell lines as compared to breast and brain tumor lines in vitro. Overall, there did not seem to be a benefit of adding IL-7. Both CD4+ and CD8+ T cells appear to mediate tumor cell apoptosis.
  
  
  CONCLUSIONS
  This study demonstrates that melanoma antigen-specific T cells can be generated from regional melanoma-draining lymph nodes and expanded ex vivo from patients with stage III disease.
authors.affiliation: Department of Surgery, Division of Surgical Oncology, University Hospitals Case Medical Center, Cleveland, OH 44106, USA.
authors.forename: Anthony
authors.initials: A
authors.lastname: Visioni
authors.forename: Mei
authors.initials: M
authors.lastname: Zhang
authors.forename: Hallie
authors.initials: H
authors.lastname: Graor
authors.forename: Julian
authors.initials: J
authors.lastname: Kim
mesh_terms: D017209:Apoptosis; D018929:Cell Culture Techniques / Q000379:methods; D015925:Cryopreservation; D006801:Humans; D016219:Immunotherapy, Adoptive / Q000379:methods*; D007376:Interleukin-2 / Q000008:administration & dosage; D015851:Interleukin-7 / Q000008:administration & dosage; D008198:Lymph Nodes / Q000276:immunology / Q000473:pathology; D008207:Lymphatic Metastasis / Q000276:immunology / Q000473:pathology; D008545:Melanoma / Q000276:immunology* / Q000473:pathology / Q000556:secondary / Q000628:therapy*; D009367:Neoplasm Staging; D013601:T-Lymphocytes / Q000276:immunology* / Q000473:pathology
chemical_list: C508594:IL2 protein, human; C508605:IL7 protein, human; D007376:Interleukin-2; D015851:Interleukin-7