title: Continuously expanding CAR NK-92 cells display selective cytotoxicity against B-cell leukemia and lymphoma.
abstract: Natural killer (NK) cells can rapidly respond to transformed and stressed cells and represent an important effector cell type for adoptive immunotherapy. In addition to donor-derived primary NK cells, continuously expanding cytotoxic cell lines such as NK-92 are being developed for clinical applications.
  
  
  
  To enhance their therapeutic utility for the treatment of B-cell malignancies, we engineered NK-92 cells by lentiviral gene transfer to express chimeric antigen receptors (CARs) that target CD19 and contain human CD3ζ (CAR 63.z), composite CD28-CD3ζ or CD137-CD3ζ signaling domains (CARs 63.28.z and 63.137.z).
  
  
  
  Exposure of CD19-positive targets to CAR NK-92 cells resulted in formation of conjugates between NK and cancer cells, NK-cell degranulation and selective cytotoxicity toward established B-cell leukemia and lymphoma cells. Likewise, the CAR NK cells displayed targeted cell killing of primary pre-B-ALL blasts that were resistant to parental NK-92. Although all three CAR NK-92 cell variants were functionally active, NK-92/63.137.z cells were less effective than NK-92/63.z and NK-92/63.28.z in cell killing and cytokine production, pointing to differential effects of the costimulatory CD28 and CD137 domains. In a Raji B-cell lymphoma model in NOD-SCID IL2R γnullmice, treatment with NK-92/63.z cells, but not parental NK-92 cells, inhibited disease progression, indicating that selective cytotoxicity was retained in vivo.
  
  
  
  Our data demonstrate that it is feasible to generate CAR-engineered NK-92 cells with potent and selective antitumor activity. These cells may become clinically useful as a continuously expandable off-the-shelf cell therapeutic agent.
authors.affiliation: Georg-Speyer-Haus, Institute for Tumor Biology and Experimental Therapy, Frankfurt, Germany.
authors.forename: Sarah
authors.initials: S
authors.lastname: Oelsner
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: Georg-Speyer-Haus, Institute for Tumor Biology and Experimental Therapy, Frankfurt, Germany.
authors.forename: Congcong
authors.initials: C
authors.lastname: Zhang
authors.affiliation: Division for Stem Cell Transplantation and Immunology, Hospital for Children and Adolescents, Goethe University Frankfurt, Frankfurt, Germany; LOEWE Center for Cell and Gene Therapy, Goethe University Frankfurt, Frankfurt, Germany.
authors.forename: Juliane
authors.initials: J
authors.lastname: Wagner
authors.affiliation: Department of Medicine, Hematology and Oncology, Goethe University Frankfurt, Frankfurt, Germany.
authors.forename: Susanne
authors.initials: S
authors.lastname: Badura
authors.affiliation: Division for Stem Cell Transplantation and Immunology, Hospital for Children and Adolescents, Goethe University Frankfurt, Frankfurt, Germany.
authors.forename: Peter
authors.initials: P
authors.lastname: Bader
authors.affiliation: Division for Stem Cell Transplantation and Immunology, Hospital for Children and Adolescents, Goethe University Frankfurt, Frankfurt, Germany; LOEWE Center for Cell and Gene Therapy, Goethe University Frankfurt, Frankfurt, Germany.
authors.forename: Evelyn
authors.initials: E
authors.lastname: Ullrich
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: NantKwest, Inc., Culver City, California, USA.
authors.forename: Hans
authors.initials: H
authors.lastname: Klingemann
authors.affiliation: Institute for Transfusion Medicine, German Red Cross Blood Donation Service North-East, Dresden and Transfusion Medicine, Medical Faculty Carl Gustav Carus, TU Dresden, Germany.
authors.forename: Torsten
authors.initials: T
authors.lastname: Tonn
authors.affiliation: Georg-Speyer-Haus, Institute for Tumor Biology and Experimental Therapy, Frankfurt, Germany; German Cancer Consortium (DKTK), partner site Frankfurt/Mainz, Germany. Electronic address: wels@gsh.uni-frankfurt.de.
authors.forename: Winfried S
authors.initials: WS
authors.lastname: Wels
mesh_terms: D000818:Animals; D018941:Antigens, CD19 / Q000276:immunology; D001402:B-Lymphocytes / Q000276:immunology; D017252:CD3 Complex / Q000235:genetics / Q000378:metabolism; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D003602:Cytotoxicity, Immunologic / Q000235:genetics; D000939:Epitopes / Q000235:genetics; D057809:HEK293 Cells; D006801:Humans; D016219:Immunotherapy, Adoptive / Q000379:methods; D007694:Killer Cells, Natural / Q000276:immunology / Q000378:metabolism / Q000637:transplantation; D015451:Leukemia, Lymphocytic, Chronic, B-Cell / Q000276:immunology / Q000628:therapy*; D008213:Lymphocyte Activation / Q000235:genetics; D008223:Lymphoma / Q000276:immunology / Q000628:therapy*; D016393:Lymphoma, B-Cell / Q000276:immunology / Q000628:therapy; D008297:Male; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D053261:Tumor Necrosis Factor Receptor Superfamily, Member 9 / Q000235:genetics / Q000378:metabolism
chemical_list: D018941:Antigens, CD19; D017252:CD3 Complex; C121704:CD3 antigen, zeta chain; D000939:Epitopes; D011993:Recombinant Fusion Proteins; D053261:Tumor Necrosis Factor Receptor Superfamily, Member 9
keywords: B-cell malignancies; CD19; adoptive immunotherapy; chimeric antigen receptor; natural killer cells