title: CD19-CAR engineered NK-92 cells are sufficient to overcome NK cell resistance in B-cell malignancies.
abstract: Many B-cell acute and chronic leukaemias tend to be resistant to killing by natural killer (NK) cells. The introduction of chimeric antigen receptors (CAR) into T cells or NK cells could potentially overcome this resistance. Here, we extend our previous observations on the resistance of malignant lymphoblasts to NK-92 cells, a continuously growing NK cell line, showing that anti-CD19-CAR (αCD19-CAR) engineered NK-92 cells can regain significant cytotoxicity against CD19 positive leukaemic cell lines and primary leukaemia cells that are resistant to cytolytic activity of parental NK-92 cells. The 'first generation' CAR was generated from a scFv (CD19) antibody fragment, coupled to a flexible hinge region, the CD3ζ chain and a Myc-tag and cloned into a retrovirus backbone. No difference in cytotoxic activity of NK-92 and transduced αCD19-CAR NK-92 cells towards CD19 negative targets was found. However, αCD19-CAR NK-92 cells specifically and efficiently lysed CD19 expressing B-precursor leukaemia cell lines as well as lymphoblasts from leukaemia patients. Since NK-92 cells can be easily expanded to clinical grade numbers under current Good Manufactoring Practice (cGMP) conditions and its safety has been documented in several phase I clinical studies, treatment with CAR modified NK-92 should be considered a treatment option for patients with lymphoid malignancies.
authors.affiliation: Department of Hematology, J.W. Goethe University Frankfurt/Main, Frankfurt, Germany.|Institute for Transfusion Medicine and Immunohematology, J.W. Goethe University Frankfurt/Main, Red Cross Blood Donor Service Baden-Württemberg-Hessen, Dresden, Germany.
authors.forename: Annette
authors.initials: A
authors.lastname: Romanski
authors.affiliation: Georg-Speyer-Haus, Institute for Tumor Biology and Experimental Therapy, Frankfurt, Germany.
authors.forename: Christoph
authors.initials: C
authors.lastname: Uherek
authors.affiliation: Department of Hematology, J.W. Goethe University Frankfurt/Main, Frankfurt, Germany.
authors.forename: Gesine
authors.initials: G
authors.lastname: Bug
authors.affiliation: Institute for Transfusion Medicine and Immunohematology, J.W. Goethe University Frankfurt/Main, Red Cross Blood Donor Service Baden-Württemberg-Hessen, Dresden, Germany.
authors.forename: Erhard
authors.initials: E
authors.lastname: Seifried
authors.affiliation: Nantkwest Inc & Tufts University Medical School, Boston, MA, USA.
authors.forename: Hans
authors.initials: H
authors.lastname: Klingemann
authors.affiliation: Georg-Speyer-Haus, Institute for Tumor Biology and Experimental Therapy, Frankfurt, Germany.
authors.forename: Winfried S
authors.initials: WS
authors.lastname: Wels
authors.affiliation: Department of Hematology, J.W. Goethe University Frankfurt/Main, Frankfurt, Germany.
authors.forename: Oliver G
authors.initials: OG
authors.lastname: Ottmann
authors.affiliation: Institute for Transfusion Medicine and Immunohematology, J.W. Goethe University Frankfurt/Main, Red Cross Blood Donor Service Baden-Württemberg-Hessen, Dresden, Germany.|Institute for Transfusion Medicine Dresden, German Red Cross Blood Donation Service North/East, Medical Faculty Carl Gustav Carus, TU Dresden, Dresden, Germany.
authors.forename: Torsten
authors.initials: T
authors.lastname: Tonn
mesh_terms: D001402:B-Lymphocytes / Q000276:immunology*; D045744:Cell Line, Tumor; D019070:Cell Lineage; D003602:Cytotoxicity, Immunologic; D005818:Genetic Engineering; D006801:Humans; D007694:Killer Cells, Natural / Q000276:immunology*; D016393:Lymphoma, B-Cell / Q000276:immunology* / Q000473:pathology; D011948:Receptors, Antigen, T-Cell / Q000378:metabolism*; D012190:Retroviridae / Q000378:metabolism; D057127:Single-Chain Antibodies / Q000378:metabolism; D014019:Tissue Donors; D014162:Transfection
chemical_list: C000605956:CD19-specific chimeric antigen receptor; D011948:Receptors, Antigen, T-Cell; D057127:Single-Chain Antibodies
keywords: CAR; NK-92; cellular immunotherapy; leukaemia; natural killer cell; αCD19