title: Vaccination with cetuximab mimotopes and biological properties of induced anti-epidermal growth factor receptor antibodies.
abstract: BACKGROUND
  The monoclonal antibody cetuximab (IMC-225, Erbitux) inhibits epidermal growth factor receptor (EGFR) signaling and has been approved for metastatic colon cancer therapy. However, to achieve effective titers, passive antibody therapies must be repeatedly administered over long periods. To overcome this limitation, we aimed to generate a vaccine inducing continuously available "cetuximab-like" antibodies in vivo using the mimotope approach.
  
  
  METHODS
  We used the phage display technique to identify four peptides structurally mimicking the cetuximab epitope. We coupled two of these peptides to an immunogenic carrier protein, and we vaccinated four groups (n = 8) of BALB/c mice intraperitoneally with 10 microg of the mimotope conjugates, a control peptide conjugate, or the carrier protein alone. We assessed antibody-dependent cellular cytotoxicity and complement-dependent cytotoxicity mediated by the induced antibodies against EGFR-overexpressing human A431 carcinoma cells. We then tested receptor internalization capacity of the induced antibodies with fluorescently labeled EGFR, and we assayed their growth inhibitory potential toward A431 cells with a [3H]thymidine proliferation assay.
  
  
  RESULTS
  Mimotope-induced antibodies recognized EGFR, and both types of antibody-mediated cytotoxic effects were elicited by these antibodies. In both cellular cytotoxicity assays, the mimotope-induced antibodies exhibited specific lysis of more than 50%. The induced antibodies caused internalization of the receptor from the cell surface into endocytic vesicles and inhibited growth of EGFR-expressing cells to a similar extent as cetuximab [67% (95% confidence interval {CI} = 55% to 79%) and 69% (95% CI = 55% to 84%), respectively].
  
  
  CONCLUSIONS
  Epitope-specific immunization is feasible for active anti-EGFR immunotherapy. The in vitro biologic features of mimotope-induced antibodies are similar to those of the monoclonal antibody cetuximab.
authors.affiliation: Department of Pathophysiology, Medical University of Vienna, Vienna, Austria.
authors.forename: Angelika B
authors.initials: AB
authors.lastname: Riemer
authors.forename: Harald
authors.initials: H
authors.lastname: Kurz
authors.forename: Markus
authors.initials: M
authors.lastname: Klinger
authors.forename: Otto
authors.initials: O
authors.lastname: Scheiner
authors.forename: Christoph C
authors.initials: CC
authors.lastname: Zielinski
authors.forename: Erika
authors.initials: E
authors.lastname: Jensen-Jarolim
mesh_terms: D000818:Animals; D019138:Antibodies, Blocking; D000911:Antibodies, Monoclonal / Q000494:pharmacology*; D061067:Antibodies, Monoclonal, Humanized; D000920:Antibody-Dependent Cell Cytotoxicity; D000970:Antineoplastic Agents / Q000494:pharmacology*; D001435:Bacteriophages; D001666:Binding Sites, Antibody; D015153:Blotting, Western; D001943:Breast Neoplasms / Q000188:drug therapy* / Q000276:immunology; D019496:Cancer Vaccines / Q000494:pharmacology*; D002294:Carcinoma, Squamous Cell / Q000188:drug therapy* / Q000276:immunology; D045744:Cell Line, Tumor; D000068818:Cetuximab; D004273:DNA, Neoplasm / Q000032:analysis; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes; D066246:ErbB Receptors / Q000037:antagonists & inhibitors*; D005260:Female; D006801:Humans; D007116:Immunization, Passive / Q000379:methods; D051379:Mice; D008807:Mice, Inbred BALB C; D008856:Microscopy, Fluorescence; D012107:Research Design; D017422:Sequence Analysis, DNA; D015398:Signal Transduction / Q000187:drug effects; D014162:Transfection; D014410:Tumor Stem Cell Assay
chemical_list: D019138:Antibodies, Blocking; D000911:Antibodies, Monoclonal; D061067:Antibodies, Monoclonal, Humanized; D000970:Antineoplastic Agents; D019496:Cancer Vaccines; D004273:DNA, Neoplasm; D000939:Epitopes; D066246:ErbB Receptors; D000068818:Cetuximab