title: Chimeric rat/human HER2 efficiently circumvents HER2 tolerance in cancer patients.
abstract: OBJECTIVE
  Despite the great success of HER2 vaccine strategies in animal models, effective clinical results have not yet been obtained. We studied the feasibility of using DNA coding for chimeric rat/human HER2 as a tool to break the unresponsiveness of T cells from patients with HER2-overexpressing tumors (HER2-CP).
  
  
  METHODS
  Dendritic cells (DCs) generated from patients with HER2-overexpressing breast (n = 28) and pancreatic (n = 16) cancer were transfected with DNA plasmids that express human HER2 or heterologous rat sequences in separate plasmids or as chimeric constructs encoding rat/human HER2 fusion proteins and used to activate autologous T cells. Activation was evaluated by IFN-γ ELISPOT assay, perforin expression, and ability to halt HER2+ tumor growth in vivo.
  
  
  RESULTS
  Specific sustained proliferation and IFN-γ production by CD4 and CD8 T cells from HER2-CP was observed after stimulation with autologous DCs transfected with chimeric rat/human HER2 plasmids. Instead, T cells from healthy donors (n = 22) could be easily stimulated with autologous DCs transfected with any human, rat, or chimeric rat/human HER2 plasmid. Chimeric HER2-transfected DCs from HER2-CP were also able to induce a sustained T-cell response that significantly hindered the in vivo growth of HER2(+) tumors. The efficacy of chimeric plasmids in overcoming tumor-induced T-cell dysfunction relies on their ability to circumvent suppressor effects exerted by regulatory T cells (Treg) and/or interleukin (IL)-10 and TGF-β1.
  
  
  CONCLUSIONS
  These results provide the proof of concept that chimeric rat/human HER2 plasmids can be used as effective vaccines for any HER2-CP with the advantage of being not limited to specific MHC. Clin Cancer Res; 20(11); 2910-21. ©2014 AACR.
authors.affiliation: Authors' Affiliations: Departments of Molecular Biotechnology and Health Sciences, University of Torino, Torino, Italy; Center for Experimental Research and Medical Studies (CERMS), AO Città della Salute e della Scienza di Torino, Torino, Italy; Immunogenetic and Transplant Biology Service, AO Città della Salute e della Scienza Torino, Italy; Division of Oncology, Subalpine OncoHematology Cancer Center (COES), AO Città della Salute e della Scienza di Torino, Torino, Italy; Department of Oncology, University of Turin, Orbassano, Italy; Department of Medical Sciences, University of Torino, Torino, Italy, Department of Molecular Cellular and Animal Biology, University of Camerino, Camerino, ItalyAuthors' Affiliations: Departments of Molecular Biotechnology and Health Sciences, University of Torino, Torino, Italy; Center for Experimental Research and Medical Studies (CERMS), AO Città della Salute e della Scienza di Torino, Torino, Italy; Immunogenetic and Transplant Biology Service, AO Città della Salute e della Scienza Torino, Italy; Division of Oncology, Subalpine OncoHematology Cancer Center (COES), AO Città della Salute e della Scienza di Torino, Torino, Italy; Department of Oncology, University of Turin, Orbassano, Italy; Department of Medical Sciences, University of Torino, Torino, Italy, Department of Molecular Cellular and Animal Biology, University of Camerino, Camerino, Italy.
authors.forename: Sergio
authors.initials: S
authors.lastname: Occhipinti
authors.affiliation: Authors' Affiliations: Departments of Molecular Biotechnology and Health Sciences, University of Torino, Torino, Italy; Center for Experimental Research and Medical Studies (CERMS), AO Città della Salute e della Scienza di Torino, Torino, Italy; Immunogenetic and Transplant Biology Service, AO Città della Salute e della Scienza Torino, Italy; Division of Oncology, Subalpine OncoHematology Cancer Center (COES), AO Città della Salute e della Scienza di Torino, Torino, Italy; Department of Oncology, University of Turin, Orbassano, Italy; Department of Medical Sciences, University of Torino, Torino, Italy, Department of Molecular Cellular and Animal Biology, University of Camerino, Camerino, ItalyAuthors' Affiliations: Departments of Molecular Biotechnology and Health Sciences, University of Torino, Torino, Italy; Center for Experimental Research and Medical Studies (CERMS), AO Città della Salute e della Scienza di Torino, Torino, Italy; Immunogenetic and Transplant Biology Service, AO Città della Salute e della Scienza Torino, Italy; Division of Oncology, Subalpine OncoHematology Cancer Center (COES), AO Città della Salute e della Scienza di Torino, Torino, Italy; Department of Oncology, University of Turin, Orbassano, Italy; Department of Medical Sciences, University of Torino, Torino, Italy, Department of Molecular Cellular and Animal Biology, University of Camerino, Camerino, Italy.
authors.forename: Laura
authors.initials: L
authors.lastname: Sponton
authors.affiliation: Authors' Affiliations: Departments of Molecular Biotechnology and Health Sciences, University of Torino, Torino, Italy; Center for Experimental Research and Medical Studies (CERMS), AO Città della Salute e della Scienza di Torino, Torino, Italy; Immunogenetic and Transplant Biology Service, AO Città della Salute e della Scienza Torino, Italy; Division of Oncology, Subalpine OncoHematology Cancer Center (COES), AO Città della Salute e della Scienza di Torino, Torino, Italy; Department of Oncology, University of Turin, Orbassano, Italy; Department of Medical Sciences, University of Torino, Torino, Italy, Department of Molecular Cellular and Animal Biology, University of Camerino, Camerino, ItalyAuthors' Affiliations: Departments of Molecular Biotechnology and Health Sciences, University of Torino, Torino, Italy; Center for Experimental Research and Medical Studies (CERMS), AO Città della Salute e della Scienza di Torino, Torino, Italy; Immunogenetic and Transplant Biology Service, AO Città della Salute e della Scienza Torino, Italy; Division of Oncology, Subalpine OncoHematology Cancer Center (COES), AO Città della Salute e della Scienza di Torino, Torino, Italy; Department of Oncology, University of Turin, Orbassano, Italy; Department of Medical Sciences, University of Torino, Torino, Italy, Department of Molecular Cellular and Animal Biology, University of Camerino, Camerino, Italy.
authors.forename: Simona
authors.initials: S
authors.lastname: Rolla
authors.affiliation: Authors' Affiliations: Departments of Molecular Biotechnology and Health Sciences, University of Torino, Torino, Italy; Center for Experimental Research and Medical Studies (CERMS), AO Città della Salute e della Scienza di Torino, Torino, Italy; Immunogenetic and Transplant Biology Service, AO Città della Salute e della Scienza Torino, Italy; Division of Oncology, Subalpine OncoHematology Cancer Center (COES), AO Città della Salute e della Scienza di Torino, Torino, Italy; Department of Oncology, University of Turin, Orbassano, Italy; Department of Medical Sciences, University of Torino, Torino, Italy, Department of Molecular Cellular and Animal Biology, University of Camerino, Camerino, Italy.
authors.forename: Cristiana
authors.initials: C
authors.lastname: Caorsi
authors.affiliation: Authors' Affiliations: Departments of Molecular Biotechnology and Health Sciences, University of Torino, Torino, Italy; Center for Experimental Research and Medical Studies (CERMS), AO Città della Salute e della Scienza di Torino, Torino, Italy; Immunogenetic and Transplant Biology Service, AO Città della Salute e della Scienza Torino, Italy; Division of Oncology, Subalpine OncoHematology Cancer Center (COES), AO Città della Salute e della Scienza di Torino, Torino, Italy; Department of Oncology, University of Turin, Orbassano, Italy; Department of Medical Sciences, University of Torino, Torino, Italy, Department of Molecular Cellular and Animal Biology, University of Camerino, Camerino, Italy.
authors.forename: Anna
authors.initials: A
authors.lastname: Novarino
authors.affiliation: Authors' Affiliations: Departments of Molecular Biotechnology and Health Sciences, University of Torino, Torino, Italy; Center for Experimental Research and Medical Studies (CERMS), AO Città della Salute e della Scienza di Torino, Torino, Italy; Immunogenetic and Transplant Biology Service, AO Città della Salute e della Scienza Torino, Italy; Division of Oncology, Subalpine OncoHematology Cancer Center (COES), AO Città della Salute e della Scienza di Torino, Torino, Italy; Department of Oncology, University of Turin, Orbassano, Italy; Department of Medical Sciences, University of Torino, Torino, Italy, Department of Molecular Cellular and Animal Biology, University of Camerino, Camerino, Italy.
authors.forename: Michela
authors.initials: M
authors.lastname: Donadio
authors.affiliation: Authors' Affiliations: Departments of Molecular Biotechnology and Health Sciences, University of Torino, Torino, Italy; Center for Experimental Research and Medical Studies (CERMS), AO Città della Salute e della Scienza di Torino, Torino, Italy; Immunogenetic and Transplant Biology Service, AO Città della Salute e della Scienza Torino, Italy; Division of Oncology, Subalpine OncoHematology Cancer Center (COES), AO Città della Salute e della Scienza di Torino, Torino, Italy; Department of Oncology, University of Turin, Orbassano, Italy; Department of Medical Sciences, University of Torino, Torino, Italy, Department of Molecular Cellular and Animal Biology, University of Camerino, Camerino, Italy.
authors.forename: Sara
authors.initials: S
authors.lastname: Bustreo
authors.affiliation: Authors' Affiliations: Departments of Molecular Biotechnology and Health Sciences, University of Torino, Torino, Italy; Center for Experimental Research and Medical Studies (CERMS), AO Città della Salute e della Scienza di Torino, Torino, Italy; Immunogenetic and Transplant Biology Service, AO Città della Salute e della Scienza Torino, Italy; Division of Oncology, Subalpine OncoHematology Cancer Center (COES), AO Città della Salute e della Scienza di Torino, Torino, Italy; Department of Oncology, University of Turin, Orbassano, Italy; Department of Medical Sciences, University of Torino, Torino, Italy, Department of Molecular Cellular and Animal Biology, University of Camerino, Camerino, Italy.
authors.forename: Maria Antonietta
authors.initials: MA
authors.lastname: Satolli
authors.affiliation: Authors' Affiliations: Departments of Molecular Biotechnology and Health Sciences, University of Torino, Torino, Italy; Center for Experimental Research and Medical Studies (CERMS), AO Città della Salute e della Scienza di Torino, Torino, Italy; Immunogenetic and Transplant Biology Service, AO Città della Salute e della Scienza Torino, Italy; Division of Oncology, Subalpine OncoHematology Cancer Center (COES), AO Città della Salute e della Scienza di Torino, Torino, Italy; Department of Oncology, University of Turin, Orbassano, Italy; Department of Medical Sciences, University of Torino, Torino, Italy, Department of Molecular Cellular and Animal Biology, University of Camerino, Camerino, Italy.
authors.forename: Carla
authors.initials: C
authors.lastname: Pecchioni
authors.affiliation: Authors' Affiliations: Departments of Molecular Biotechnology and Health Sciences, University of Torino, Torino, Italy; Center for Experimental Research and Medical Studies (CERMS), AO Città della Salute e della Scienza di Torino, Torino, Italy; Immunogenetic and Transplant Biology Service, AO Città della Salute e della Scienza Torino, Italy; Division of Oncology, Subalpine OncoHematology Cancer Center (COES), AO Città della Salute e della Scienza di Torino, Torino, Italy; Department of Oncology, University of Turin, Orbassano, Italy; Department of Medical Sciences, University of Torino, Torino, Italy, Department of Molecular Cellular and Animal Biology, University of Camerino, Camerino, Italy.
authors.forename: Cristina
authors.initials: C
authors.lastname: Marchini
authors.affiliation: Authors' Affiliations: Departments of Molecular Biotechnology and Health Sciences, University of Torino, Torino, Italy; Center for Experimental Research and Medical Studies (CERMS), AO Città della Salute e della Scienza di Torino, Torino, Italy; Immunogenetic and Transplant Biology Service, AO Città della Salute e della Scienza Torino, Italy; Division of Oncology, Subalpine OncoHematology Cancer Center (COES), AO Città della Salute e della Scienza di Torino, Torino, Italy; Department of Oncology, University of Turin, Orbassano, Italy; Department of Medical Sciences, University of Torino, Torino, Italy, Department of Molecular Cellular and Animal Biology, University of Camerino, Camerino, Italy.
authors.forename: Augusto
authors.initials: A
authors.lastname: Amici
authors.affiliation: Authors' Affiliations: Departments of Molecular Biotechnology and Health Sciences, University of Torino, Torino, Italy; Center for Experimental Research and Medical Studies (CERMS), AO Città della Salute e della Scienza di Torino, Torino, Italy; Immunogenetic and Transplant Biology Service, AO Città della Salute e della Scienza Torino, Italy; Division of Oncology, Subalpine OncoHematology Cancer Center (COES), AO Città della Salute e della Scienza di Torino, Torino, Italy; Department of Oncology, University of Turin, Orbassano, Italy; Department of Medical Sciences, University of Torino, Torino, Italy, Department of Molecular Cellular and Animal Biology, University of Camerino, Camerino, Italy.
authors.forename: Federica
authors.initials: F
authors.lastname: Cavallo
authors.affiliation: Authors' Affiliations: Departments of Molecular Biotechnology and Health Sciences, University of Torino, Torino, Italy; Center for Experimental Research and Medical Studies (CERMS), AO Città della Salute e della Scienza di Torino, Torino, Italy; Immunogenetic and Transplant Biology Service, AO Città della Salute e della Scienza Torino, Italy; Division of Oncology, Subalpine OncoHematology Cancer Center (COES), AO Città della Salute e della Scienza di Torino, Torino, Italy; Department of Oncology, University of Turin, Orbassano, Italy; Department of Medical Sciences, University of Torino, Torino, Italy, Department of Molecular Cellular and Animal Biology, University of Camerino, Camerino, ItalyAuthors' Affiliations: Departments of Molecular Biotechnology and Health Sciences, University of Torino, Torino, Italy; Center for Experimental Research and Medical Studies (CERMS), AO Città della Salute e della Scienza di Torino, Torino, Italy; Immunogenetic and Transplant Biology Service, AO Città della Salute e della Scienza Torino, Italy; Division of Oncology, Subalpine OncoHematology Cancer Center (COES), AO Città della Salute e della Scienza di Torino, Torino, Italy; Department of Oncology, University of Turin, Orbassano, Italy; Department of Medical Sciences, University of Torino, Torino, Italy, Department of Molecular Cellular and Animal Biology, University of Camerino, Camerino, Italy.
authors.forename: Paola
authors.initials: P
authors.lastname: Cappello
authors.affiliation: Authors' Affiliations: Departments of Molecular Biotechnology and Health Sciences, University of Torino, Torino, Italy; Center for Experimental Research and Medical Studies (CERMS), AO Città della Salute e della Scienza di Torino, Torino, Italy; Immunogenetic and Transplant Biology Service, AO Città della Salute e della Scienza Torino, Italy; Division of Oncology, Subalpine OncoHematology Cancer Center (COES), AO Città della Salute e della Scienza di Torino, Torino, Italy; Department of Oncology, University of Turin, Orbassano, Italy; Department of Medical Sciences, University of Torino, Torino, Italy, Department of Molecular Cellular and Animal Biology, University of Camerino, Camerino, ItalyAuthors' Affiliations: Departments of Molecular Biotechnology and Health Sciences, University of Torino, Torino, Italy; Center for Experimental Research and Medical Studies (CERMS), AO Città della Salute e della Scienza di Torino, Torino, Italy; Immunogenetic and Transplant Biology Service, AO Città della Salute e della Scienza Torino, Italy; Division of Oncology, Subalpine OncoHematology Cancer Center (COES), AO Città della Salute e della Scienza di Torino, Torino, Italy; Department of Oncology, University of Turin, Orbassano, Italy; Department of Medical Sciences, University of Torino, Torino, Italy, Department of Molecular Cellular and Animal Biology, University of Camerino, Camerino, Italy.
authors.forename: Daniele
authors.initials: D
authors.lastname: Pierobon
authors.affiliation: Authors' Affiliations: Departments of Molecular Biotechnology and Health Sciences, University of Torino, Torino, Italy; Center for Experimental Research and Medical Studies (CERMS), AO Città della Salute e della Scienza di Torino, Torino, Italy; Immunogenetic and Transplant Biology Service, AO Città della Salute e della Scienza Torino, Italy; Division of Oncology, Subalpine OncoHematology Cancer Center (COES), AO Città della Salute e della Scienza di Torino, Torino, Italy; Department of Oncology, University of Turin, Orbassano, Italy; Department of Medical Sciences, University of Torino, Torino, Italy, Department of Molecular Cellular and Animal Biology, University of Camerino, Camerino, ItalyAuthors' Affiliations: Departments of Molecular Biotechnology and Health Sciences, University of Torino, Torino, Italy; Center for Experimental Research and Medical Studies (CERMS), AO Città della Salute e della Scienza di Torino, Torino, Italy; Immunogenetic and Transplant Biology Service, AO Città della Salute e della Scienza Torino, Italy; Division of Oncology, Subalpine OncoHematology Cancer Center (COES), AO Città della Salute e della Scienza di Torino, Torino, Italy; Department of Oncology, University of Turin, Orbassano, Italy; Department of Medical Sciences, University of Torino, Torino, Italy, Department of Molecular Cellular and Animal Biology, University of Camerino, Camerino, Italy.
authors.forename: Francesco
authors.initials: F
authors.lastname: Novelli
authors.affiliation: Authors' Affiliations: Departments of Molecular Biotechnology and Health Sciences, University of Torino, Torino, Italy; Center for Experimental Research and Medical Studies (CERMS), AO Città della Salute e della Scienza di Torino, Torino, Italy; Immunogenetic and Transplant Biology Service, AO Città della Salute e della Scienza Torino, Italy; Division of Oncology, Subalpine OncoHematology Cancer Center (COES), AO Città della Salute e della Scienza di Torino, Torino, Italy; Department of Oncology, University of Turin, Orbassano, Italy; Department of Medical Sciences, University of Torino, Torino, Italy, Department of Molecular Cellular and Animal Biology, University of Camerino, Camerino, ItalyAuthors' Affiliations: Departments of Molecular Biotechnology and Health Sciences, University of Torino, Torino, Italy; Center for Experimental Research and Medical Studies (CERMS), AO Città della Salute e della Scienza di Torino, Torino, Italy; Immunogenetic and Transplant Biology Service, AO Città della Salute e della Scienza Torino, Italy; Division of Oncology, Subalpine OncoHematology Cancer Center (COES), AO Città della Salute e della Scienza di Torino, Torino, Italy; Department of Oncology, University of Turin, Orbassano, Italy; Department of Medical Sciences, University of Torino, Torino, Italy, Department of Molecular Cellular and Animal Biology, University of Camerino, Camerino, Italy mirella.giovarelli@unito.it.
authors.forename: Mirella
authors.initials: M
authors.lastname: Giovarelli
mesh_terms: D000818:Animals; D001943:Breast Neoplasms / Q000276:immunology*; D019496:Cancer Vaccines / Q000276:immunology* / Q000494:pharmacology; D003713:Dendritic Cells / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D007108:Immune Tolerance / Q000276:immunology; D007150:Immunohistochemistry; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D010190:Pancreatic Neoplasms / Q000276:immunology*; D010957:Plasmids; D051381:Rats; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000276:immunology / Q000494:pharmacology; D014162:Transfection; D018183:Transplantation Chimera; D019444:Vaccines, DNA / Q000276:immunology* / Q000494:pharmacology; D023041:Xenograft Model Antitumor Assays
chemical_list: D019496:Cancer Vaccines; D018719:Erb-b2 Receptor Tyrosine Kinases; D011993:Recombinant Fusion Proteins; D019444:Vaccines, DNA