title: A cell surface-displayed anti-c-myc single-chain antibody: new perspectives for the genetic improvement of cellular tumor vaccines.
abstract: We have shown recently that cell surface-bound, single-chain Fv antibodies (scFv) are a powerful tool for the improvement of cellular tumor vaccines. To simplify this approach and to develop a general tool for the generation and improvement of cellular tumor vaccines, we chose an scFv against a peptide from the human proto-oncogene c-myc that could anchor any c-myc-tagged protein to the cell surface. The retroviral vector p50-Mx-neo (pMESV) was used to express scFv on the surface of the human melanoma line SkMel63. The cell-bound anti-c-myc scFv bound specifically to a soluble purified anti-CD28 scFv carrying a c-myc peptide-tag at its C terminus. Proof of principle was determined by incubating human peripheral blood lymphocytes with a mixture of (a) anti-c-myc-transfected SkMel63 cells binding the anti-CD28 scFv and (b) SkMel63 cells transfected with an anti-CD3 scFv. A clear synergistic effect on T-cell activation was observed that was comparable with that obtained in previous studies using SkMel63 cells transfected with the gene for the anti-CD28 scFv. As the cell surface-displayed anti-c-myc scFv can bind any c-myc-tagged protein of interest, this technique facilitates the genetic engineering of cellular vaccines for the therapy of virtually all human neoplasias.
authors.affiliation: Recombinant Antibody Group, Experimental Therapy and Diagnosis Programme, German Cancer Research Center, Heidelberg.
authors.forename: C
authors.initials: C
authors.lastname: de Inés
authors.forename: B
authors.initials: B
authors.lastname: Cochlovius
authors.forename: S
authors.initials: S
authors.lastname: Schmidt
authors.forename: M
authors.initials: M
authors.lastname: Little
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology*; D000954:Antigens, Surface / Q000276:immunology*; D001665:Binding Sites; D019496:Cancer Vaccines / Q000627:therapeutic use*; D002455:Cell Division / Q000187:drug effects; D017931:DNA Primers / Q000737:chemistry; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D015870:Gene Expression; D016259:Genes, myc / Q000235:genetics / Q000276:immunology*; D005822:Genetic Vectors; D006801:Humans; D007125:Immunogenetics; D007128:Immunoglobulin Fragments / Q000276:immunology*; D007371:Interferon-gamma / Q000032:analysis; D015847:Interleukin-4 / Q000032:analysis; D008213:Lymphocyte Activation / Q000276:immunology*; D016133:Polymerase Chain Reaction; D000090063:Proto-Oncogene Mas; D013601:T-Lymphocytes / Q000276:immunology*; D014407:Tumor Cells, Cultured / Q000187:drug effects
chemical_list: D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D019496:Cancer Vaccines; D017931:DNA Primers; D007128:Immunoglobulin Fragments; C000715470:MAS1 protein, human; D000090063:Proto-Oncogene Mas; D015847:Interleukin-4; D007371:Interferon-gamma