title: Characterization of a sustained-release delivery system for combined cytokine/peptide vaccination using a poly-N-acetyl glucosamine-based polymer matrix.
abstract: Identification of tumor-associated antigens (TAAs) and their class I MHC-restricted epitopes now allows for the rational design of peptide-based cancer vaccines. A biocompatible system capable of sustained release of biologically relevant levels of cytokine and TAA peptide could provide a more effective microenvironment for antigen presentation. Our goal was to test a sustained-release cytokine/TAA peptide-based formulation using a highly purified polysaccharide [poly-N-acetyl glucosamine (p-GlcNAc)] polymer. Granulocyte-macrophage colony-stimulating factor (GM-CSF; 100 microgram) and MART-1(27-35) peptide (128 microgram in DMSO) were formulated into p-GlcNAc. Peptide release was assayed in vitro using interleukin 2 production from previously characterized MART-1(27-35)-specific Jurkat T cells (JRT22). GM-CSF release was assayed via ELISA and proliferation of M-07e (GM-CSF-dependent) cells. Local bioavailability of MART-1(27-35) peptide for uptake and presentation by antigen-presenting cells was demonstrated for up to 6 days (>0.5 microgram/ml). More than 1.0 microgram/ml GM-CSF was concomitantly released over the same period. Biocompatibility and local tissue response to p-GlcNAc releasing murine GM-CSF was determined in C57BL/6 mice via s.c. injection using murine GM-CSF (0. 2 microgram/ml) in 200 microliter of a 2.5% polymer gel. Significant lymphocytic and eosinophilic infiltration was observed 2-7 days after injection with polymer containing murine GM-CSF. The results of our studies show that this biocompatible system is capable of a sustained concomitant release of biologically active peptide and cytokine into the local microenvironment. These findings support further studies to validate a p-GlcNAc delivery system vehicle for a cytokine/TAA peptide-based cancer vaccine.
authors.affiliation: Departments of Surgery, (Division of Hematology/Oncology), University of South Carolina, Charleston, South Carolina.
authors.forename: D J
authors.initials: DJ
authors.lastname: Cole
authors.forename: S
authors.initials: S
authors.lastname: Gattoni-Celli
authors.forename: E F
authors.initials: EF
authors.lastname: McClay
authors.forename: J S
authors.initials: JS
authors.lastname: Metcalf
authors.forename: J M
authors.initials: JM
authors.lastname: Brown
authors.forename: N
authors.initials: N
authors.lastname: Nabavi
authors.forename: D A
authors.initials: DA
authors.lastname: Newton
authors.forename: C B
authors.initials: CB
authors.lastname: Woolhiser
authors.forename: M C
authors.initials: MC
authors.lastname: Wilson
authors.forename: J N
authors.initials: JN
authors.lastname: Vournakis
mesh_terms: D000117:Acetylglucosamine; D000818:Animals; D000951:Antigens, Neoplasm / Q000008:administration & dosage* / Q000378:metabolism; D001672:Biocompatible Materials; D019496:Cancer Vaccines / Q000008:administration & dosage*; D016207:Cytokines / Q000008:administration & dosage / Q000493:pharmacokinetics; D003692:Delayed-Action Preparations; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000008:administration & dosage* / Q000493:pharmacokinetics*; D006801:Humans; D019169:Jurkat Cells; D058965:MART-1 Antigen; D051379:Mice; D008810:Mice, Inbred C57BL; D009363:Neoplasm Proteins / Q000008:administration & dosage* / Q000493:pharmacokinetics*; D010446:Peptide Fragments / Q000008:administration & dosage* / Q000493:pharmacokinetics; D011134:Polysaccharides; D011994:Recombinant Proteins / Q000008:administration & dosage / Q000493:pharmacokinetics
chemical_list: D000951:Antigens, Neoplasm; D001672:Biocompatible Materials; D019496:Cancer Vaccines; D016207:Cytokines; D003692:Delayed-Action Preparations; D058965:MART-1 Antigen; C548667:MLANA protein, human; C548668:Mlana protein, mouse; D009363:Neoplasm Proteins; D010446:Peptide Fragments; D011134:Polysaccharides; D011994:Recombinant Proteins; C113831:poly-N-acetyl glucosamine; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D000117:Acetylglucosamine