title: Modification of Expanded NK Cells with Chimeric Antigen Receptor mRNA for Adoptive Cellular Therapy.
abstract: NK cells are bone marrow-derived cytotoxic lymphocytes that play a major role in the rejection of tumors and cells infected by viruses. The regulation of NK activation vs inhibition is regulated by the expression of a variety of NK receptors (NKRs) and specific NKRs' ligands expressed on their targets. However, factors limiting NK therapy include small numbers of active NK cells in unexpanded peripheral blood and lack of specific tumor targeting. Chimeric antigen receptors (CAR) usually include a single-chain Fv variable fragment from a monoclonal antibody, a transmembrane hinge region, and a signaling domain such as CD28, CD3-zeta, 4-1BB (CD137), or 2B4 (CD244) endodimers. Redirecting NK cells with a CAR will circumvent the limitations of the lack of NK targeting specificity. This chapter focuses on the methods to expand human NK cells from peripheral blood by co-culturing with feeder cells and to modify the expanded NK cells efficiently with the in vitro transcribed CAR mRNA by electroporation and to test the functionality of the CAR-modified expanded NK cells for use in adoptive cellular immunotherapy.
authors.affiliation: Department of Pediatrics, New York Medical College, Valhalla, NY, 10595, USA.
authors.forename: Yaya
authors.initials: Y
authors.lastname: Chu
authors.affiliation: Department of Pediatrics, New York Medical College, Valhalla, NY, 10595, USA.
authors.forename: Allyson
authors.initials: A
authors.lastname: Flower
authors.affiliation: Department of Pediatrics, New York Medical College, Valhalla, NY, 10595, USA. mitchell_cairo@nymc.edu.|Department of Medicine, Maria Fareri Children's Hospital at Westchester Medical Center, New York Medical College, Valhalla, NY, 10595, USA. mitchell_cairo@nymc.edu.|Department of Pathology, New York Medical College, Valhalla, NY, 10595, USA. mitchell_cairo@nymc.edu.|Department of Microbiology and Immunology, New York Medical College, Valhalla, NY, 10595, USA. mitchell_cairo@nymc.edu.|Department of Cell Biology and Anatomy, New York Medical College, Valhalla, NY, 10595, USA. mitchell_cairo@nymc.edu.|Maria Fareri Children's Hospital at Westchester Medical Center, New York Medical College, 40 Sunshine Cottage Rd. Skyline Office 1N-D12, Valhalla, NY, 10595, USA. mitchell_cairo@nymc.edu.
authors.forename: Mitchell S
authors.initials: MS
authors.lastname: Cairo
mesh_terms: D045744:Cell Line, Tumor; D018920:Coculture Techniques / Q000379:methods*; D018274:Electroporation; D061252:Feeder Cells / Q000166:cytology / Q000276:immunology; D006801:Humans; D016219:Immunotherapy, Adoptive; D007694:Killer Cells, Natural / Q000166:cytology* / Q000276:immunology; D007962:Leukocytes / Q000166:cytology; D008213:Lymphocyte Activation; D012333:RNA, Messenger / Q000378:metabolism; D011946:Receptors, Antigen / Q000235:genetics*
chemical_list: D012333:RNA, Messenger; D011946:Receptors, Antigen
keywords: Adoptive cell therapy; Chimeric antigen receptor; Electroporation; Natural killer cells; mRNA