title: Selective bispecific T cell recruiting antibody and antitumor activity of adoptive T cell transfer.
abstract: BACKGROUND
  One bottleneck for adoptive T cell therapy (ACT) is recruitment of T cells into tumors. We hypothesized that combining tumor-specific T cells, modified with a marker antigen and a bispecific antibody (BiAb) that selectively recognizes transduced T cells and tumor cells would improve T cell recruitment to tumors and enhance therapeutic efficacy.
  
  
  METHODS
  SV40 T antigen-specific T cells from T cell receptor (TCR)-I-transgenic mice were transduced with a truncated human epidermal growth factor receptor (EGFR) as a marker protein. Targeting and killing by combined ACT and anti-EGFR-anti-EpCAM BiAb therapy was analyzed in C57Bl/6 mice (n = six to 12 per group) carrying subcutaneous tumors of the murine gastric cancer cell line GC8 (SV40(+) and EpCAM(+)). Anti-EGFR x anti-c-Met BiAb was used for targeting of human tumor-specific T cells to c-Met(+) human tumor cell lines. Differences between experimental conditions were analyzed using the Student's t test, and differences in tumor growth with two-way analysis of variance. Overall survival was analyzed by log-rank test. All statistical tests were two-sided.
  
  
  RESULTS
  The BiAb linked EGFR-transduced T cells to tumor cells and enhanced tumor cell lysis. In vivo, the combination of ACT and Biab produced increased T cell infiltration of tumors, retarded tumor growth, and prolonged survival compared with ACT with a control antibody (median survival 95 vs 75 days, P < .001). In human cells, this strategy enhanced recruitment of human EGFR-transduced T cells to immobilized c-Met and recognition of tyrosinase(+) melanoma cells by TCR-, as well as of CEA(+) colon cancer cells by chimeric antigen receptor (CAR)-modified T cells.
  
  
  CONCLUSIONS
  BiAb recruitment of tumor-specific T cells transduced with a marker antigen to tumor cells may enhance efficacy of ACT.
authors.affiliation: Center of Integrated Protein Science Munich (CIPS-M) and Division of Clinical Pharmacology, Department of Internal Medicine IV, Ludwig-Maximilians-Universität München, Munich, Germany (SK, JS, MiC, SG, JH, YZ, JCS, MS, SR, CB, SE); Roche Pharmaceutical Research and Early Development, Oncology Discovery and Translational Area, Roche Innovation Center Penzberg, Penzberg, Germany (RC, CS, GN); Center for Molecular Medicine Cologne and Department I for Internal Medicine, University Hospital Cologne, Cologne, Germany (MaC, HA); Institute of Molecular Immunology, Helmholtz Zentrum München and Clinical Cooperation Group Immune Monitoring, Helmholtz Zentrum München, German Research Center for Environmental Health, Munich, Germany (DJS); Roche Pharmaceutical Research and Early Development, Oncology Discovery and Translational Area, Roche Innovation Center Zurich, Switzerland (CL); Chair of Pharmacology, Department of Medicine, University of Fribourg, Fribourg, Switzerland (CB). sebastian.kobold@med.uni-muenchen.de.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Kobold
authors.affiliation: Center of Integrated Protein Science Munich (CIPS-M) and Division of Clinical Pharmacology, Department of Internal Medicine IV, Ludwig-Maximilians-Universität München, Munich, Germany (SK, JS, MiC, SG, JH, YZ, JCS, MS, SR, CB, SE); Roche Pharmaceutical Research and Early Development, Oncology Discovery and Translational Area, Roche Innovation Center Penzberg, Penzberg, Germany (RC, CS, GN); Center for Molecular Medicine Cologne and Department I for Internal Medicine, University Hospital Cologne, Cologne, Germany (MaC, HA); Institute of Molecular Immunology, Helmholtz Zentrum München and Clinical Cooperation Group Immune Monitoring, Helmholtz Zentrum München, German Research Center for Environmental Health, Munich, Germany (DJS); Roche Pharmaceutical Research and Early Development, Oncology Discovery and Translational Area, Roche Innovation Center Zurich, Switzerland (CL); Chair of Pharmacology, Department of Medicine, University of Fribourg, Fribourg, Switzerland (CB).
authors.forename: Julius
authors.initials: J
authors.lastname: Steffen
authors.affiliation: Center of Integrated Protein Science Munich (CIPS-M) and Division of Clinical Pharmacology, Department of Internal Medicine IV, Ludwig-Maximilians-Universität München, Munich, Germany (SK, JS, MiC, SG, JH, YZ, JCS, MS, SR, CB, SE); Roche Pharmaceutical Research and Early Development, Oncology Discovery and Translational Area, Roche Innovation Center Penzberg, Penzberg, Germany (RC, CS, GN); Center for Molecular Medicine Cologne and Department I for Internal Medicine, University Hospital Cologne, Cologne, Germany (MaC, HA); Institute of Molecular Immunology, Helmholtz Zentrum München and Clinical Cooperation Group Immune Monitoring, Helmholtz Zentrum München, German Research Center for Environmental Health, Munich, Germany (DJS); Roche Pharmaceutical Research and Early Development, Oncology Discovery and Translational Area, Roche Innovation Center Zurich, Switzerland (CL); Chair of Pharmacology, Department of Medicine, University of Fribourg, Fribourg, Switzerland (CB).
authors.forename: Michael
authors.initials: M
authors.lastname: Chaloupka
authors.affiliation: Center of Integrated Protein Science Munich (CIPS-M) and Division of Clinical Pharmacology, Department of Internal Medicine IV, Ludwig-Maximilians-Universität München, Munich, Germany (SK, JS, MiC, SG, JH, YZ, JCS, MS, SR, CB, SE); Roche Pharmaceutical Research and Early Development, Oncology Discovery and Translational Area, Roche Innovation Center Penzberg, Penzberg, Germany (RC, CS, GN); Center for Molecular Medicine Cologne and Department I for Internal Medicine, University Hospital Cologne, Cologne, Germany (MaC, HA); Institute of Molecular Immunology, Helmholtz Zentrum München and Clinical Cooperation Group Immune Monitoring, Helmholtz Zentrum München, German Research Center for Environmental Health, Munich, Germany (DJS); Roche Pharmaceutical Research and Early Development, Oncology Discovery and Translational Area, Roche Innovation Center Zurich, Switzerland (CL); Chair of Pharmacology, Department of Medicine, University of Fribourg, Fribourg, Switzerland (CB).
authors.forename: Simon
authors.initials: S
authors.lastname: Grassmann
authors.affiliation: Center of Integrated Protein Science Munich (CIPS-M) and Division of Clinical Pharmacology, Department of Internal Medicine IV, Ludwig-Maximilians-Universität München, Munich, Germany (SK, JS, MiC, SG, JH, YZ, JCS, MS, SR, CB, SE); Roche Pharmaceutical Research and Early Development, Oncology Discovery and Translational Area, Roche Innovation Center Penzberg, Penzberg, Germany (RC, CS, GN); Center for Molecular Medicine Cologne and Department I for Internal Medicine, University Hospital Cologne, Cologne, Germany (MaC, HA); Institute of Molecular Immunology, Helmholtz Zentrum München and Clinical Cooperation Group Immune Monitoring, Helmholtz Zentrum München, German Research Center for Environmental Health, Munich, Germany (DJS); Roche Pharmaceutical Research and Early Development, Oncology Discovery and Translational Area, Roche Innovation Center Zurich, Switzerland (CL); Chair of Pharmacology, Department of Medicine, University of Fribourg, Fribourg, Switzerland (CB).
authors.forename: Jonas
authors.initials: J
authors.lastname: Henkel
authors.affiliation: Center of Integrated Protein Science Munich (CIPS-M) and Division of Clinical Pharmacology, Department of Internal Medicine IV, Ludwig-Maximilians-Universität München, Munich, Germany (SK, JS, MiC, SG, JH, YZ, JCS, MS, SR, CB, SE); Roche Pharmaceutical Research and Early Development, Oncology Discovery and Translational Area, Roche Innovation Center Penzberg, Penzberg, Germany (RC, CS, GN); Center for Molecular Medicine Cologne and Department I for Internal Medicine, University Hospital Cologne, Cologne, Germany (MaC, HA); Institute of Molecular Immunology, Helmholtz Zentrum München and Clinical Cooperation Group Immune Monitoring, Helmholtz Zentrum München, German Research Center for Environmental Health, Munich, Germany (DJS); Roche Pharmaceutical Research and Early Development, Oncology Discovery and Translational Area, Roche Innovation Center Zurich, Switzerland (CL); Chair of Pharmacology, Department of Medicine, University of Fribourg, Fribourg, Switzerland (CB).
authors.forename: Raffaella
authors.initials: R
authors.lastname: Castoldi
authors.affiliation: Center of Integrated Protein Science Munich (CIPS-M) and Division of Clinical Pharmacology, Department of Internal Medicine IV, Ludwig-Maximilians-Universität München, Munich, Germany (SK, JS, MiC, SG, JH, YZ, JCS, MS, SR, CB, SE); Roche Pharmaceutical Research and Early Development, Oncology Discovery and Translational Area, Roche Innovation Center Penzberg, Penzberg, Germany (RC, CS, GN); Center for Molecular Medicine Cologne and Department I for Internal Medicine, University Hospital Cologne, Cologne, Germany (MaC, HA); Institute of Molecular Immunology, Helmholtz Zentrum München and Clinical Cooperation Group Immune Monitoring, Helmholtz Zentrum München, German Research Center for Environmental Health, Munich, Germany (DJS); Roche Pharmaceutical Research and Early Development, Oncology Discovery and Translational Area, Roche Innovation Center Zurich, Switzerland (CL); Chair of Pharmacology, Department of Medicine, University of Fribourg, Fribourg, Switzerland (CB).
authors.forename: Yi
authors.initials: Y
authors.lastname: Zeng
authors.affiliation: Center of Integrated Protein Science Munich (CIPS-M) and Division of Clinical Pharmacology, Department of Internal Medicine IV, Ludwig-Maximilians-Universität München, Munich, Germany (SK, JS, MiC, SG, JH, YZ, JCS, MS, SR, CB, SE); Roche Pharmaceutical Research and Early Development, Oncology Discovery and Translational Area, Roche Innovation Center Penzberg, Penzberg, Germany (RC, CS, GN); Center for Molecular Medicine Cologne and Department I for Internal Medicine, University Hospital Cologne, Cologne, Germany (MaC, HA); Institute of Molecular Immunology, Helmholtz Zentrum München and Clinical Cooperation Group Immune Monitoring, Helmholtz Zentrum München, German Research Center for Environmental Health, Munich, Germany (DJS); Roche Pharmaceutical Research and Early Development, Oncology Discovery and Translational Area, Roche Innovation Center Zurich, Switzerland (CL); Chair of Pharmacology, Department of Medicine, University of Fribourg, Fribourg, Switzerland (CB).
authors.forename: Markus
authors.initials: M
authors.lastname: Chmielewski
authors.affiliation: Center of Integrated Protein Science Munich (CIPS-M) and Division of Clinical Pharmacology, Department of Internal Medicine IV, Ludwig-Maximilians-Universität München, Munich, Germany (SK, JS, MiC, SG, JH, YZ, JCS, MS, SR, CB, SE); Roche Pharmaceutical Research and Early Development, Oncology Discovery and Translational Area, Roche Innovation Center Penzberg, Penzberg, Germany (RC, CS, GN); Center for Molecular Medicine Cologne and Department I for Internal Medicine, University Hospital Cologne, Cologne, Germany (MaC, HA); Institute of Molecular Immunology, Helmholtz Zentrum München and Clinical Cooperation Group Immune Monitoring, Helmholtz Zentrum München, German Research Center for Environmental Health, Munich, Germany (DJS); Roche Pharmaceutical Research and Early Development, Oncology Discovery and Translational Area, Roche Innovation Center Zurich, Switzerland (CL); Chair of Pharmacology, Department of Medicine, University of Fribourg, Fribourg, Switzerland (CB).
authors.forename: Jan C
authors.initials: JC
authors.lastname: Schmollinger
authors.affiliation: Center of Integrated Protein Science Munich (CIPS-M) and Division of Clinical Pharmacology, Department of Internal Medicine IV, Ludwig-Maximilians-Universität München, Munich, Germany (SK, JS, MiC, SG, JH, YZ, JCS, MS, SR, CB, SE); Roche Pharmaceutical Research and Early Development, Oncology Discovery and Translational Area, Roche Innovation Center Penzberg, Penzberg, Germany (RC, CS, GN); Center for Molecular Medicine Cologne and Department I for Internal Medicine, University Hospital Cologne, Cologne, Germany (MaC, HA); Institute of Molecular Immunology, Helmholtz Zentrum München and Clinical Cooperation Group Immune Monitoring, Helmholtz Zentrum München, German Research Center for Environmental Health, Munich, Germany (DJS); Roche Pharmaceutical Research and Early Development, Oncology Discovery and Translational Area, Roche Innovation Center Zurich, Switzerland (CL); Chair of Pharmacology, Department of Medicine, University of Fribourg, Fribourg, Switzerland (CB).
authors.forename: Max
authors.initials: M
authors.lastname: Schnurr
authors.affiliation: Center of Integrated Protein Science Munich (CIPS-M) and Division of Clinical Pharmacology, Department of Internal Medicine IV, Ludwig-Maximilians-Universität München, Munich, Germany (SK, JS, MiC, SG, JH, YZ, JCS, MS, SR, CB, SE); Roche Pharmaceutical Research and Early Development, Oncology Discovery and Translational Area, Roche Innovation Center Penzberg, Penzberg, Germany (RC, CS, GN); Center for Molecular Medicine Cologne and Department I for Internal Medicine, University Hospital Cologne, Cologne, Germany (MaC, HA); Institute of Molecular Immunology, Helmholtz Zentrum München and Clinical Cooperation Group Immune Monitoring, Helmholtz Zentrum München, German Research Center for Environmental Health, Munich, Germany (DJS); Roche Pharmaceutical Research and Early Development, Oncology Discovery and Translational Area, Roche Innovation Center Zurich, Switzerland (CL); Chair of Pharmacology, Department of Medicine, University of Fribourg, Fribourg, Switzerland (CB).
authors.forename: Simon
authors.initials: S
authors.lastname: Rothenfußer
authors.affiliation: Center of Integrated Protein Science Munich (CIPS-M) and Division of Clinical Pharmacology, Department of Internal Medicine IV, Ludwig-Maximilians-Universität München, Munich, Germany (SK, JS, MiC, SG, JH, YZ, JCS, MS, SR, CB, SE); Roche Pharmaceutical Research and Early Development, Oncology Discovery and Translational Area, Roche Innovation Center Penzberg, Penzberg, Germany (RC, CS, GN); Center for Molecular Medicine Cologne and Department I for Internal Medicine, University Hospital Cologne, Cologne, Germany (MaC, HA); Institute of Molecular Immunology, Helmholtz Zentrum München and Clinical Cooperation Group Immune Monitoring, Helmholtz Zentrum München, German Research Center for Environmental Health, Munich, Germany (DJS); Roche Pharmaceutical Research and Early Development, Oncology Discovery and Translational Area, Roche Innovation Center Zurich, Switzerland (CL); Chair of Pharmacology, Department of Medicine, University of Fribourg, Fribourg, Switzerland (CB).
authors.forename: Dolores J
authors.initials: DJ
authors.lastname: Schendel
authors.affiliation: Center of Integrated Protein Science Munich (CIPS-M) and Division of Clinical Pharmacology, Department of Internal Medicine IV, Ludwig-Maximilians-Universität München, Munich, Germany (SK, JS, MiC, SG, JH, YZ, JCS, MS, SR, CB, SE); Roche Pharmaceutical Research and Early Development, Oncology Discovery and Translational Area, Roche Innovation Center Penzberg, Penzberg, Germany (RC, CS, GN); Center for Molecular Medicine Cologne and Department I for Internal Medicine, University Hospital Cologne, Cologne, Germany (MaC, HA); Institute of Molecular Immunology, Helmholtz Zentrum München and Clinical Cooperation Group Immune Monitoring, Helmholtz Zentrum München, German Research Center for Environmental Health, Munich, Germany (DJS); Roche Pharmaceutical Research and Early Development, Oncology Discovery and Translational Area, Roche Innovation Center Zurich, Switzerland (CL); Chair of Pharmacology, Department of Medicine, University of Fribourg, Fribourg, Switzerland (CB).
authors.forename: Hinrich
authors.initials: H
authors.lastname: Abken
authors.affiliation: Center of Integrated Protein Science Munich (CIPS-M) and Division of Clinical Pharmacology, Department of Internal Medicine IV, Ludwig-Maximilians-Universität München, Munich, Germany (SK, JS, MiC, SG, JH, YZ, JCS, MS, SR, CB, SE); Roche Pharmaceutical Research and Early Development, Oncology Discovery and Translational Area, Roche Innovation Center Penzberg, Penzberg, Germany (RC, CS, GN); Center for Molecular Medicine Cologne and Department I for Internal Medicine, University Hospital Cologne, Cologne, Germany (MaC, HA); Institute of Molecular Immunology, Helmholtz Zentrum München and Clinical Cooperation Group Immune Monitoring, Helmholtz Zentrum München, German Research Center for Environmental Health, Munich, Germany (DJS); Roche Pharmaceutical Research and Early Development, Oncology Discovery and Translational Area, Roche Innovation Center Zurich, Switzerland (CL); Chair of Pharmacology, Department of Medicine, University of Fribourg, Fribourg, Switzerland (CB).
authors.forename: Claudio
authors.initials: C
authors.lastname: Sustmann
authors.affiliation: Center of Integrated Protein Science Munich (CIPS-M) and Division of Clinical Pharmacology, Department of Internal Medicine IV, Ludwig-Maximilians-Universität München, Munich, Germany (SK, JS, MiC, SG, JH, YZ, JCS, MS, SR, CB, SE); Roche Pharmaceutical Research and Early Development, Oncology Discovery and Translational Area, Roche Innovation Center Penzberg, Penzberg, Germany (RC, CS, GN); Center for Molecular Medicine Cologne and Department I for Internal Medicine, University Hospital Cologne, Cologne, Germany (MaC, HA); Institute of Molecular Immunology, Helmholtz Zentrum München and Clinical Cooperation Group Immune Monitoring, Helmholtz Zentrum München, German Research Center for Environmental Health, Munich, Germany (DJS); Roche Pharmaceutical Research and Early Development, Oncology Discovery and Translational Area, Roche Innovation Center Zurich, Switzerland (CL); Chair of Pharmacology, Department of Medicine, University of Fribourg, Fribourg, Switzerland (CB).
authors.forename: Gerhard
authors.initials: G
authors.lastname: Niederfellner
authors.affiliation: Center of Integrated Protein Science Munich (CIPS-M) and Division of Clinical Pharmacology, Department of Internal Medicine IV, Ludwig-Maximilians-Universität München, Munich, Germany (SK, JS, MiC, SG, JH, YZ, JCS, MS, SR, CB, SE); Roche Pharmaceutical Research and Early Development, Oncology Discovery and Translational Area, Roche Innovation Center Penzberg, Penzberg, Germany (RC, CS, GN); Center for Molecular Medicine Cologne and Department I for Internal Medicine, University Hospital Cologne, Cologne, Germany (MaC, HA); Institute of Molecular Immunology, Helmholtz Zentrum München and Clinical Cooperation Group Immune Monitoring, Helmholtz Zentrum München, German Research Center for Environmental Health, Munich, Germany (DJS); Roche Pharmaceutical Research and Early Development, Oncology Discovery and Translational Area, Roche Innovation Center Zurich, Switzerland (CL); Chair of Pharmacology, Department of Medicine, University of Fribourg, Fribourg, Switzerland (CB).
authors.forename: Christian
authors.initials: C
authors.lastname: Klein
authors.affiliation: Center of Integrated Protein Science Munich (CIPS-M) and Division of Clinical Pharmacology, Department of Internal Medicine IV, Ludwig-Maximilians-Universität München, Munich, Germany (SK, JS, MiC, SG, JH, YZ, JCS, MS, SR, CB, SE); Roche Pharmaceutical Research and Early Development, Oncology Discovery and Translational Area, Roche Innovation Center Penzberg, Penzberg, Germany (RC, CS, GN); Center for Molecular Medicine Cologne and Department I for Internal Medicine, University Hospital Cologne, Cologne, Germany (MaC, HA); Institute of Molecular Immunology, Helmholtz Zentrum München and Clinical Cooperation Group Immune Monitoring, Helmholtz Zentrum München, German Research Center for Environmental Health, Munich, Germany (DJS); Roche Pharmaceutical Research and Early Development, Oncology Discovery and Translational Area, Roche Innovation Center Zurich, Switzerland (CL); Chair of Pharmacology, Department of Medicine, University of Fribourg, Fribourg, Switzerland (CB).
authors.forename: Carole
authors.initials: C
authors.lastname: Bourquin
authors.affiliation: Center of Integrated Protein Science Munich (CIPS-M) and Division of Clinical Pharmacology, Department of Internal Medicine IV, Ludwig-Maximilians-Universität München, Munich, Germany (SK, JS, MiC, SG, JH, YZ, JCS, MS, SR, CB, SE); Roche Pharmaceutical Research and Early Development, Oncology Discovery and Translational Area, Roche Innovation Center Penzberg, Penzberg, Germany (RC, CS, GN); Center for Molecular Medicine Cologne and Department I for Internal Medicine, University Hospital Cologne, Cologne, Germany (MaC, HA); Institute of Molecular Immunology, Helmholtz Zentrum München and Clinical Cooperation Group Immune Monitoring, Helmholtz Zentrum München, German Research Center for Environmental Health, Munich, Germany (DJS); Roche Pharmaceutical Research and Early Development, Oncology Discovery and Translational Area, Roche Innovation Center Zurich, Switzerland (CL); Chair of Pharmacology, Department of Medicine, University of Fribourg, Fribourg, Switzerland (CB).
authors.forename: Stefan
authors.initials: S
authors.lastname: Endres
mesh_terms: D019264:Adoptive Transfer; D000704:Analysis of Variance; D000818:Animals; D000951:Antigens, Neoplasm / Q000276:immunology / Q000378:metabolism*; D015815:Cell Adhesion Molecules / Q000276:immunology / Q000378:metabolism*; D045744:Cell Line, Tumor; D000071858:Epithelial Cell Adhesion Molecule; D066246:ErbB Receptors / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D019859:Proto-Oncogene Proteins c-met / Q000276:immunology / Q000378:metabolism*; D011948:Receptors, Antigen, T-Cell / Q000378:metabolism*; D013274:Stomach Neoplasms / Q000276:immunology* / Q000628:therapy*; D013601:T-Lymphocytes / Q000276:immunology*; D014161:Transduction, Genetic
chemical_list: D000951:Antigens, Neoplasm; D015815:Cell Adhesion Molecules; C499379:EPCAM protein, human; D000071858:Epithelial Cell Adhesion Molecule; D011948:Receptors, Antigen, T-Cell; C512478:EGFR protein, human; D066246:ErbB Receptors; D019859:Proto-Oncogene Proteins c-met