title: Biolistic DNA vaccination against melanoma.
abstract: We describe here the use of particle-mediated gene transfer for the induction of immune responses against melanoma antigens in murine tumor models using the melanocyte differentiation antigen tyrosinase-related protein 2 (TRP2) as an antigen in a murine B16 melanoma model. We have utilized marker genes such as β-galactosidase (βgal) and EGFP, which can be readily detected, as control antigens to establish the gene delivery and to detect antigen-specific humoral and cellular immune responses. After biolistic DNA vaccination with plasmids encoding the TRP2 gene we observed the induction of TRP2-specific T-cells and antibodies associated with vitiligo-like fur depigmentation and tumor immunity against B16 melanoma cells. Here we describe the preparation of cartridges with DNA-coated gold beads and the in vivo gene transfer into skin using the Helios Gene Gun system. We also describe protocols for the measurement of humoral and cellular immune responses against the melanocyte differentiation antigen TRP2. These protocols can subsequently be adapted to other antigens.
authors.affiliation: Institute for Laboratory Animal Science, University Hospital of RWTH Aachen, Aachen, Germany. jsteitz@ukaachen.de
authors.forename: Julia
authors.initials: J
authors.lastname: Steitz
authors.forename: Thomas
authors.initials: T
authors.lastname: Tüting
mesh_terms: D000818:Animals; D000918:Antibody Specificity; D000951:Antigens, Neoplasm / Q000276:immunology; D019470:Biolistics / Q000295:instrumentation*; D015153:Blotting, Western; D019496:Cancer Vaccines / Q000008:administration & dosage* / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D005260:Female; D006046:Gold / Q000737:chemistry; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D007371:Interferon-gamma / Q000378:metabolism; D019746:Intramolecular Oxidoreductases / Q000276:immunology; D008175:Lung Neoplasms / Q000235:genetics / Q000276:immunology / Q000556:secondary; D008546:Melanoma, Experimental / Q000235:genetics / Q000517:prevention & control*; D051379:Mice; D010858:Pigmentation / Q000276:immunology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology; D014611:Vaccination / Q000295:instrumentation*; D019444:Vaccines, DNA / Q000008:administration & dosage* / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D001616:beta-Galactosidase / Q000235:genetics
chemical_list: D000951:Antigens, Neoplasm; D019496:Cancer Vaccines; D019444:Vaccines, DNA; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D006046:Gold; D007371:Interferon-gamma; D001616:beta-Galactosidase; D019746:Intramolecular Oxidoreductases; C042501:dopachrome isomerase