Anti-metastatic immunotherapy based on mucosal administration of flagellin and immunomodulatory P10

Immunol Cell Biol. 2015 Jan;93(1):86-98. doi: 10.1038/icb.2014.74. Epub 2014 Sep 16.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Administration, Intranasal
  • Administration, Mucosal
  • Animals
  • Antigens, Bacterial / administration & dosage
  • Antigens, Bacterial / genetics
  • Antigens, Bacterial / immunology*
  • Cancer Vaccines / administration & dosage
  • Cancer Vaccines / genetics
  • Cancer Vaccines / immunology*
  • Caspase 1 / deficiency
  • Caspase 1 / genetics
  • Flagellin / administration & dosage
  • Flagellin / genetics
  • Flagellin / immunology*
  • Gene Expression
  • Glycoproteins / administration & dosage
  • Glycoproteins / genetics
  • Glycoproteins / immunology*
  • Immunotherapy / methods*
  • Injections, Intravenous
  • Interferon-gamma / agonists
  • Interferon-gamma / biosynthesis
  • Interferon-gamma / immunology
  • Interleukin-12 Subunit p40 / biosynthesis
  • Interleukin-12 Subunit p40 / immunology
  • Lung / drug effects
  • Lung / immunology
  • Lung / pathology
  • Lung Neoplasms / genetics
  • Lung Neoplasms / immunology
  • Lung Neoplasms / pathology
  • Lung Neoplasms / therapy*
  • Lymph Nodes / drug effects
  • Lymph Nodes / immunology
  • Lymph Nodes / pathology
  • Macrophages / drug effects
  • Macrophages / immunology
  • Macrophages / pathology
  • Male
  • Melanoma, Experimental / genetics
  • Melanoma, Experimental / immunology
  • Melanoma, Experimental / pathology
  • Melanoma, Experimental / therapy*
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Myeloid Differentiation Factor 88 / genetics
  • Myeloid Differentiation Factor 88 / immunology
  • Neoplasm Metastasis
  • Peptide Fragments / administration & dosage
  • Peptide Fragments / genetics
  • Peptide Fragments / immunology*
  • Toll-Like Receptor 5 / agonists
  • Toll-Like Receptor 5 / genetics
  • Toll-Like Receptor 5 / immunology

Substances

  • Antigens, Bacterial
  • Cancer Vaccines
  • Glycoproteins
  • Interleukin-12 Subunit p40
  • Myd88 protein, mouse
  • Myeloid Differentiation Factor 88
  • P10 peptide, Paracoccidioides brasiliensis
  • Peptide Fragments
  • Toll-Like Receptor 5
  • Flagellin
  • Interferon-gamma
  • Caspase 1