title: Anti-metastatic immunotherapy based on mucosal administration of flagellin and immunomodulatory P10.
abstract: Current therapies against malignant melanoma generally fail to increase survival in most patients, and immunotherapy is a promising approach as it could reduce the dosage of toxic therapeutic drugs. In the present study, we show that an immunotherapeutic approach based on the use of the Toll-like receptor (TLR)-5 ligand flagellin (Salmonella Typhimurium FliCi) combined with the major histocompatibility complex class II-restricted P10 peptide, derived from the Paracoccidioides brasiliensis gp43 major surface protein, reduced the number of lung metastasis in a murine melanoma model. Compounds were administered intranasally into C57Bl/6 mice intravenously challenged with syngeneic B16F10-Nex2 melanoma cells, aiming at the local (pulmonary) immune response modulation. Along with a marked reduction in the number of lung nodules, a significant increase in survival was observed. The immunization regimen induced both local and systemic proinflammatory responses. Lung macrophages were polarized towards a M1 phenotype, lymph node cells, and splenocytes secreted higher interleukin-12p40 and interferon (IFN)-γ levels when re-stimulated with tumor antigens. The protective effect of the FliCi+P10 formulation required TLR-5, myeloid differentiation primary response gene 88 and IFN-γ expression, but caspase-1 knockout mice were only partially protected, suggesting that intracellular flagellin receptors are not involved with the anti-tumor effect. The immune therapy resulted in the activation of tumor-specific CD4(+) T lymphocytes, which conferred protection to metastatic melanoma growth after adoptive transfer. Taken together, our results report a new immunotherapeutic approach based on TLR-5 activation and IFN-γ production capable to control the metastatic growth of B16F10-Nex2 melanoma, being a promising alternative to be associated with chemotherapeutic drugs for an effective anti-tumor responses.
authors.affiliation: Laboratory of Cancer Immunobiology, Department of Microbiology, Immunology and Parasitology, Escola Paulista de Medicina-Universidade Federal de São Paulo (EPM-UNIFESP), São Paulo, Brazil.
authors.forename: Filipe M
authors.initials: FM
authors.lastname: de Melo
authors.affiliation: Vaccine Development Laboratory, Department of Microbiology, Institute of Biomedical Sciences, University of São Paulo (ICB-USP), São Paulo, Brazil.
authors.forename: Catarina J M
authors.initials: CJ
authors.lastname: Braga
authors.affiliation: Laboratory of Cancer Immunobiology, Department of Microbiology, Immunology and Parasitology, Escola Paulista de Medicina-Universidade Federal de São Paulo (EPM-UNIFESP), São Paulo, Brazil.
authors.forename: Felipe V
authors.initials: FV
authors.lastname: Pereira
authors.affiliation: Laboratory of Retrovirology, Department of Microbiology, Immunology and Parasitology, EPM-UNIFESP, São Paulo, Brazil.
authors.forename: Juliana T
authors.initials: JT
authors.lastname: Maricato
authors.affiliation: Laboratory of Transplantation Immunobiology, Department of Immunology, ICB-USP, São Paulo, Brazil.
authors.forename: Clarice S T
authors.initials: CS
authors.lastname: Origassa
authors.affiliation: Laboratory of Cancer Immunobiology, Department of Microbiology, Immunology and Parasitology, Escola Paulista de Medicina-Universidade Federal de São Paulo (EPM-UNIFESP), São Paulo, Brazil.
authors.forename: Mariana F
authors.initials: MF
authors.lastname: Souza
authors.affiliation: Laboratory of Cancer Immunobiology, Department of Microbiology, Immunology and Parasitology, Escola Paulista de Medicina-Universidade Federal de São Paulo (EPM-UNIFESP), São Paulo, Brazil.
authors.forename: Amanda C
authors.initials: AC
authors.lastname: Melo
authors.affiliation: Laboratory of Cancer Immunobiology, Department of Microbiology, Immunology and Parasitology, Escola Paulista de Medicina-Universidade Federal de São Paulo (EPM-UNIFESP), São Paulo, Brazil.
authors.forename: Priscila
authors.initials: P
authors.lastname: Silva
authors.affiliation: Laboratory of Cancer Immunobiology, Department of Microbiology, Immunology and Parasitology, Escola Paulista de Medicina-Universidade Federal de São Paulo (EPM-UNIFESP), São Paulo, Brazil.
authors.forename: Samanta L
authors.initials: SL
authors.lastname: Tomaz
authors.affiliation: Laboratory of Cancer Immunobiology, Department of Microbiology, Immunology and Parasitology, Escola Paulista de Medicina-Universidade Federal de São Paulo (EPM-UNIFESP), São Paulo, Brazil.
authors.forename: Karina P
authors.initials: KP
authors.lastname: Gimenes
authors.affiliation: Experimental Oncology Unit (UNONEX), Department of Microbiology, Immunology and Parasitology, (EPM-UNIFESP), São Paulo, Brazil.
authors.forename: Jorge A B
authors.initials: JA
authors.lastname: Scutti
authors.affiliation: Department of Biophysics, EPM-UNIFESP, São Paulo, Brazil.
authors.forename: Maria A
authors.initials: MA
authors.lastname: Juliano
authors.affiliation: Department of Cellular and Molecular Biology and Pathogenic Bioagents, School of Medicine of Ribeirão Preto, University of São Paulo, Ribeirão Preto, Brazil.
authors.forename: Dario S
authors.initials: DS
authors.lastname: Zamboni
authors.affiliation: Laboratory of Transplantation Immunobiology, Department of Immunology, ICB-USP, São Paulo, Brazil.
authors.forename: Niels O
authors.initials: NO
authors.lastname: Câmara
authors.affiliation: Experimental Oncology Unit (UNONEX), Department of Microbiology, Immunology and Parasitology, (EPM-UNIFESP), São Paulo, Brazil.
authors.forename: Luiz R
authors.initials: LR
authors.lastname: Travassos
authors.affiliation: Vaccine Development Laboratory, Department of Microbiology, Institute of Biomedical Sciences, University of São Paulo (ICB-USP), São Paulo, Brazil.
authors.forename: Luis C S
authors.initials: LC
authors.lastname: Ferreira
authors.affiliation: Laboratory of Cancer Immunobiology, Department of Microbiology, Immunology and Parasitology, Escola Paulista de Medicina-Universidade Federal de São Paulo (EPM-UNIFESP), São Paulo, Brazil.
authors.forename: Elaine G
authors.initials: EG
authors.lastname: Rodrigues
mesh_terms: D000281:Administration, Intranasal; D058956:Administration, Mucosal; D000818:Animals; D000942:Antigens, Bacterial / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D019496:Cancer Vaccines / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D020170:Caspase 1 / Q000172:deficiency / Q000235:genetics; D005408:Flagellin / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D015870:Gene Expression; D006023:Glycoproteins / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D007167:Immunotherapy / Q000379:methods*; D007275:Injections, Intravenous; D007371:Interferon-gamma / Q000819:agonists / Q000096:biosynthesis / Q000276:immunology; D053762:Interleukin-12 Subunit p40 / Q000096:biosynthesis / Q000276:immunology; D008168:Lung / Q000187:drug effects / Q000276:immunology / Q000473:pathology; D008175:Lung Neoplasms / Q000235:genetics / Q000276:immunology / Q000473:pathology / Q000628:therapy*; D008198:Lymph Nodes / Q000187:drug effects / Q000276:immunology / Q000473:pathology; D008264:Macrophages / Q000187:drug effects / Q000276:immunology / Q000473:pathology; D008297:Male; D008546:Melanoma, Experimental / Q000235:genetics / Q000276:immunology / Q000473:pathology / Q000628:therapy*; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D053594:Myeloid Differentiation Factor 88 / Q000235:genetics / Q000276:immunology; D009362:Neoplasm Metastasis; D010446:Peptide Fragments / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D051198:Toll-Like Receptor 5 / Q000819:agonists / Q000235:genetics / Q000276:immunology
chemical_list: D000942:Antigens, Bacterial; D019496:Cancer Vaccines; D006023:Glycoproteins; D053762:Interleukin-12 Subunit p40; C507780:Myd88 protein, mouse; D053594:Myeloid Differentiation Factor 88; C550731:P10 peptide, Paracoccidioides brasiliensis; D010446:Peptide Fragments; D051198:Toll-Like Receptor 5; D005408:Flagellin; D007371:Interferon-gamma; D020170:Caspase 1