title: A phase I clinical study of vaccination of melanoma patients with dendritic cells loaded with allogeneic apoptotic/necrotic melanoma cells. Analysis of toxicity and immune response to the vaccine and of IL-10 -1082 promoter genotype as predictor of disease progression.
abstract: BACKGROUND
  Sixteen melanoma patients (1 stage IIC, 8 stage III, and 7 stage IV) were treated in a Phase I study with a vaccine (DC/Apo-Nec) composed of autologous dendritic cells (DCs) loaded with a mixture of apoptotic/necrotic allogeneic melanoma cell lines (Apo-Nec), to evaluate toxicity and immune responses. Also, IL-10 1082 genotype was analyzed in an effort to predict disease progression.
  
  
  METHODS
  PBMC were obtained after leukapheresis and DCs were generated from monocytes cultured in the presence of GM-CSF and IL-4 in serum-free medium. Immature DCs were loaded with gamma-irradiated Apo-Nec cells and injected id without adjuvant. Cohorts of four patients were given four vaccines each with 5, 10, 15, or 20 x 106 DC/Apo-Nec cell per vaccine, two weeks apart. Immune responses were measured by ELISpot and tetramer analysis. Il-10 genotype was measured by PCR and corroborated by IL-10 production by stimulated PBMC.
  
  
  RESULTS
  Immature DCs efficiently phagocytosed melanoma Apo-Nec cells and matured after phagocytosis as evidenced by increased expression of CD83, CD80, CD86, HLA class I and II, and 75.2 +/- 16% reduction in Dextran-FITC endocytosis. CCR7 was also up-regulated upon Apo-Nec uptake in DCs from all patients, and accordingly DC/Apo-Nec cells were able to migrate in vitro toward MIP-3 beta. The vaccine was well tolerated in all patients. The DTH score increased significantly in all patients after the first vaccination (Mann-Whitney Test, p < 0.05). The presence of CD8+T lymphocytes specific to gp100 and Melan A/MART-1 Ags was determined by ELISpot and tetramer analysis in five HLA-A*0201 patients before and after vaccination; one patient had stable elevated levels before and after vaccination; two increased their CD8 + levels, one had stable moderate and one had negligible levels. The analysis of IL-10 promoter -1082 polymorphism in the sixteen patients showed a positive correlation between AA genotype, accompanied by lower in vitro IL-10 production by stimulated PBMC, and faster melanoma progression after lymph nodes surgery (p = 0.04). With a mean follow-up of 49.5 months post-surgery, one stage IIC patient and 7/8 stage III patients remain NED but 7/7 stage IV patients have progressed.
  
  
  CONCLUSIONS
  We conclude that DC/Apo-Nec vaccine is safe, well tolerated and it may induce specific immunity against melanoma Ags. Patients with a low-producing IL-10 polymorphism appear to have a worst prognosis.
  
  
  BACKGROUND
  Clinicaltrials.gov (NHI) NCT00515983.
authors.affiliation: Centro de Investigaciones Oncológicas FUCA, Cramer 1180, (1426) Buenos Aires, Argentina. evoneuw@leloir.org.ar
authors.forename: Erika M
authors.initials: EM
authors.lastname: von Euw
authors.forename: María M
authors.initials: MM
authors.lastname: Barrio
authors.forename: David
authors.initials: D
authors.lastname: Furman
authors.forename: Estrella M
authors.initials: EM
authors.lastname: Levy
authors.forename: Michele
authors.initials: M
authors.lastname: Bianchini
authors.forename: Isabelle
authors.initials: I
authors.lastname: Peguillet
authors.forename: Olivier
authors.initials: O
authors.lastname: Lantz
authors.forename: Alejandra
authors.initials: A
authors.lastname: Vellice
authors.forename: Abraham
authors.initials: A
authors.lastname: Kohan
authors.forename: Matías
authors.initials: M
authors.lastname: Chacón
authors.forename: Cassian
authors.initials: C
authors.lastname: Yee
authors.forename: Rosa
authors.initials: R
authors.lastname: Wainstok
authors.forename: José
authors.initials: J
authors.lastname: Mordoh
mesh_terms: D000293:Adolescent; D000328:Adult; D017209:Apoptosis; D019496:Cancer Vaccines / Q000633:toxicity*; D003713:Dendritic Cells / Q000276:immunology* / Q000637:transplantation*; D018450:Disease Progression; D005260:Female; D005838:Genotype; D006801:Humans; D006968:Hypersensitivity, Delayed; D016753:Interleukin-10 / Q000235:genetics*; D008213:Lymphocyte Activation; D008297:Male; D008545:Melanoma / Q000276:immunology* / Q000473:pathology*; D008875:Middle Aged; D009336:Necrosis; D009367:Neoplasm Staging; D018579:Patient Selection; D010587:Phagocytosis; D011110:Polymorphism, Genetic; D011237:Predictive Value of Tests; D011401:Promoter Regions, Genetic; D014182:Transplantation, Autologous
chemical_list: D019496:Cancer Vaccines; D016753:Interleukin-10