title: Vaccination using melanoma cells treated with p19arf and interferon beta gene transfer in a mouse model: a novel combination for cancer immunotherapy.
abstract: Previously, we combined p19(Arf) (Cdkn2a, tumor suppressor protein) and interferon beta (IFN-β, immunomodulatory cytokine) gene transfer in order to enhance cell death in a murine model of melanoma. Here, we present evidence of the immune response induced when B16 cells succumbing to death due to treatment with p19(Arf) and IFN-β are applied in vaccine models. Use of dying cells for prophylactic vaccination was investigated, identifying conditions for tumor-free survival. After combined p19(Arf) and IFN-β treatment, we observed immune rejection at the vaccine site in immune competent and nude mice with normal NK activity, but not in NOD-SCID and dexamethasone immunosuppressed mice (NK deficient). Combined treatment induced IL-15, ULBP1, FAS/APO1 and KILLER/DR5 expression, providing a mechanism for NK activation. Prophylactic vaccination protected against tumor challenge, where markedly delayed progression and leukocyte infiltration were observed. Analysis of primed lymphocytes revealed secretion of TH1-related cytokines and depletion protocols showed that both CD4(+) and CD8(+) T lymphocytes are necessary for immune protection. However, application of this prophylactic vaccine where cells were treated either with IFN-β alone or combined with p19(Arf) conferred similar immune protection and cytokine activation, yet only the combination was associated with increased overall survival. In a therapeutic vaccine protocol, only the combination was associated with reduced tumor progression. Our results indicate that by harnessing cell death in an immunogenic context, our p19(Arf) and IFN-β combination offers a clear advantage when both genes are included in the vaccine and warrants further development as a novel immunotherapy for melanoma.
authors.affiliation: Viral Vector Laboratory, Centro de Investigação Translacional em Oncologia/LIM24, Instituto do Câncer do Estado de São Paulo, Faculdade de Medicina da Universidade de São Paulo, Av. Dr. Arnaldo, 251, 8th Floor, São Paulo, SP, 01246-000, Brazil.
authors.forename: Ruan Felipe Vieira
authors.initials: RF
authors.lastname: Medrano
authors.affiliation: Viral Vector Laboratory, Centro de Investigação Translacional em Oncologia/LIM24, Instituto do Câncer do Estado de São Paulo, Faculdade de Medicina da Universidade de São Paulo, Av. Dr. Arnaldo, 251, 8th Floor, São Paulo, SP, 01246-000, Brazil.
authors.forename: João Paulo Portela
authors.initials: JP
authors.lastname: Catani
authors.affiliation: Viral Vector Laboratory, Centro de Investigação Translacional em Oncologia/LIM24, Instituto do Câncer do Estado de São Paulo, Faculdade de Medicina da Universidade de São Paulo, Av. Dr. Arnaldo, 251, 8th Floor, São Paulo, SP, 01246-000, Brazil.
authors.forename: Aline Hunger
authors.initials: AH
authors.lastname: Ribeiro
authors.affiliation: Escola Paulista de Medicina-Universidade Federal de São Paulo, São Paulo, Brazil.
authors.forename: Samanta Lopes
authors.initials: SL
authors.lastname: Tomaz
authors.affiliation: Animal Care Facility, Faculdade de Medicina da Universidade de São Paulo, São Paulo, Brazil.
authors.forename: Christian A
authors.initials: CA
authors.lastname: Merkel
authors.affiliation: Gene Therapy Laboratory, Instituto de Ciências Biomédicas-I, Universidade de São Paulo, São Paulo, Brazil.
authors.forename: Eugenia
authors.initials: E
authors.lastname: Costanzi-Strauss
authors.affiliation: Viral Vector Laboratory, Centro de Investigação Translacional em Oncologia/LIM24, Instituto do Câncer do Estado de São Paulo, Faculdade de Medicina da Universidade de São Paulo, Av. Dr. Arnaldo, 251, 8th Floor, São Paulo, SP, 01246-000, Brazil. bstrauss@usp.br.
authors.forename: Bryan E
authors.initials: BE
authors.lastname: Strauss
mesh_terms: D000818:Animals; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology / Q000378:metabolism; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology / Q000378:metabolism; D045744:Cell Line, Tumor; D019941:Cyclin-Dependent Kinase Inhibitor p16 / Q000235:genetics / Q000276:immunology*; D005260:Female; D015316:Genetic Therapy / Q000379:methods; D007167:Immunotherapy / Q000379:methods; D016899:Interferon-beta / Q000235:genetics / Q000276:immunology*; D019409:Interleukin-15 / Q000276:immunology / Q000378:metabolism; D007694:Killer Cells, Natural / Q000276:immunology / Q000378:metabolism; D008546:Melanoma, Experimental / Q000235:genetics / Q000276:immunology* / Q000628:therapy*; D008810:Mice, Inbred C57BL; D016688:Mice, Inbred NOD; D008819:Mice, Nude; D016513:Mice, SCID; D053220:Receptors, TNF-Related Apoptosis-Inducing Ligand / Q000276:immunology / Q000378:metabolism; D018417:Th1 Cells / Q000276:immunology / Q000378:metabolism; D047368:Tumor Burden / Q000235:genetics / Q000276:immunology; D014611:Vaccination / Q000379:methods*
chemical_list: D019941:Cyclin-Dependent Kinase Inhibitor p16; D019409:Interleukin-15; D053220:Receptors, TNF-Related Apoptosis-Inducing Ligand; D016899:Interferon-beta
keywords: Adenovirus; Cell death; Immunotherapy; Interferon beta; Melanoma; p19Arf