title: Chimeric antigen receptor-engineered cytokine-induced killer cells overcome treatment resistance of pre-B-cell acute lymphoblastic leukemia and enhance survival.
abstract: Pre-emptive cancer immunotherapy by donor lymphocyte infusion (DLI) using cytokine-induced killer (CIK) cells may be beneficial to prevent relapse with a reduced risk of causing graft-versus-host-disease. CIK cells are a heterogeneous effector cell population including T cells (CD3(+) CD56(-) ), natural killer (NK) cells (CD3(-) CD56(+) ) and natural killer T (T-NK) cells (CD3(+) CD56(+) ) that exhibit non-major histocompatibility complex (MHC)-restricted cytotoxicity and are generated by ex vivo expansion of peripheral blood mononuclear cells in the presence of interferon (IFN)-γ, anti-CD3 antibody, interleukin-2 (IL-2) and interleukin-15 (IL-15). To facilitate selective target-cell recognition and enhance specific cytotoxicity against B-cell acute lymphoblastic leukemia (B-ALL), we transduced CIK cells with a lentiviral vector encoding a chimeric antigen receptor (CAR) that carries a composite CD28-CD3ζ domain for signaling and a CD19-specific scFv antibody fragment for cell binding (CAR 63.28.z). In vitro analysis revealed high and specific cell killing activity of CD19-targeted CIK/63.28.z cells against otherwise CIK-resistant cancer cell lines and primary B-ALL blasts, which was dependent on CD19 expression and CAR signaling. In a xenograft model in immunodeficient mice, treatment with CIK/63.28.z cells in contrast to therapy with unmodified CIK cells resulted in complete and durable molecular remissions of established primary pre-B-ALL. Our results demonstrate potent antileukemic activity of CAR-engineered CIK cells in vitro and in vivo, and suggest this strategy as a promising approach for adoptive immunotherapy of refractory pre-B-ALL.
authors.affiliation: Georg-Speyer-Haus, Institute for Tumor Biology and Experimental Therapy, Frankfurt, Germany.|Division for Stem Cell Transplantation and Immunology, Hospital for Children and Adolescents, Goethe University, Frankfurt, Germany.
authors.forename: Sarah
authors.initials: S
authors.lastname: Oelsner
authors.affiliation: Division for Stem Cell Transplantation and Immunology, Hospital for Children and Adolescents, Goethe University, Frankfurt, Germany.|LOEWE Center for Cell and Gene Therapy, Goethe University, Frankfurt, Germany.
authors.forename: Juliane
authors.initials: J
authors.lastname: Wagner
authors.affiliation: Georg-Speyer-Haus, Institute for Tumor Biology and Experimental Therapy, Frankfurt, Germany.
authors.forename: Miriam E
authors.initials: ME
authors.lastname: Friede
authors.affiliation: Division for Stem Cell Transplantation and Immunology, Hospital for Children and Adolescents, Goethe University, Frankfurt, Germany.
authors.forename: Verena
authors.initials: V
authors.lastname: Pfirrmann
authors.affiliation: Georg-Speyer-Haus, Institute for Tumor Biology and Experimental Therapy, Frankfurt, Germany.
authors.forename: Sabrina
authors.initials: S
authors.lastname: Genßler
authors.affiliation: Division for Stem Cell Transplantation and Immunology, Hospital for Children and Adolescents, Goethe University, Frankfurt, Germany.
authors.forename: Eva
authors.initials: E
authors.lastname: Rettinger
authors.affiliation: Molecular Biotechnology and Gene Therapy, Paul-Ehrlich-Institut, Langen, Germany.|German Cancer Consortium (DKTK), Heidelberg, Germany.
authors.forename: Christian J
authors.initials: CJ
authors.lastname: Buchholz
authors.affiliation: Department of Medicine, Hematology and Oncology, Goethe University, Frankfurt, Germany.
authors.forename: Heike
authors.initials: H
authors.lastname: Pfeifer
authors.affiliation: Division for Allergology, Pneumology and Cystic Fibrosis, Hospital for Children and Adolescents, Goethe University, Frankfurt, Germany.
authors.forename: Ralf
authors.initials: R
authors.lastname: Schubert
authors.affiliation: Department of Haematology, Division of Cancer and Genetics, Cardiff University School of Medicine, Cardiff, United Kingdom.
authors.forename: Oliver G
authors.initials: OG
authors.lastname: Ottmann
authors.affiliation: Division for Stem Cell Transplantation and Immunology, Hospital for Children and Adolescents, Goethe University, Frankfurt, Germany.|LOEWE Center for Cell and Gene Therapy, Goethe University, Frankfurt, Germany.
authors.forename: Evelyn
authors.initials: E
authors.lastname: Ullrich
authors.affiliation: Division for Stem Cell Transplantation and Immunology, Hospital for Children and Adolescents, Goethe University, Frankfurt, Germany.
authors.forename: Peter
authors.initials: P
authors.lastname: Bader
authors.affiliation: Georg-Speyer-Haus, Institute for Tumor Biology and Experimental Therapy, Frankfurt, Germany.|German Cancer Consortium (DKTK), Partner Site Frankfurt/Mainz, Germany.
authors.forename: Winfried S
authors.initials: WS
authors.lastname: Wels
mesh_terms: D000818:Animals; D015703:Antigens, CD / Q000235:genetics / Q000276:immunology; D060846:Cell Engineering / Q000379:methods; D045744:Cell Line, Tumor; D055612:Cytokine-Induced Killer Cells / Q000276:immunology* / Q000637:transplantation; D005260:Female; D057809:HEK293 Cells; D006801:Humans; D007128:Immunoglobulin Fragments / Q000235:genetics / Q000276:immunology; D016219:Immunotherapy, Adoptive / Q000379:methods*; D008297:Male; D051379:Mice; D016513:Mice, SCID; D054198:Precursor Cell Lymphoblastic Leukemia-Lymphoma / Q000276:immunology / Q000628:therapy*; D011948:Receptors, Antigen, T-Cell / Q000235:genetics / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology*; D014161:Transduction, Genetic; D023041:Xenograft Model Antitumor Assays
chemical_list: D015703:Antigens, CD; D007128:Immunoglobulin Fragments; D011948:Receptors, Antigen, T-Cell; D011993:Recombinant Fusion Proteins
keywords: B-ALL; CD19; adoptive immunotherapy; chimeric antigen receptor; cytokine-induced killer cells