title: Elutriated lymphocytes for manufacturing chimeric antigen receptor T cells.
abstract: Clinical trials of Chimeric Antigen Receptor (CAR) T cells manufactured from autologous peripheral blood mononuclear cell (PBMC) concentrates for the treatment of hematologic malignancies have been promising, but CAR T cell yields have been variable. This variability is due in part to the contamination of the PBMC concentrates with monocytes and granulocytes.
  
  
  
  Counter-flow elutriation allows for the closed system separation of lymphocytes from monocytes and granulocytes. We investigated the use of PBMC concentrates enriched for lymphocytes using elutriation for manufacturing 8 CD19- and 5 GD2-CAR T cell products.
  
  
  
  When compared to PBMC concentrates, lymphocyte-enriched elutriation fractions contained greater proportions of CD3+ and CD56+ cells and reduced proportions of CD14+ and CD15+ cells. All 13 CAR T cell products manufactured using the elutriated lymphocytes yielded sufficient quantities of transduced CAR T cells to meet clinical dose criteria. The GD2-CAR T cell products contained significantly more T cells and transduced T cells than the CD19-CAR T cell products. A comparison of the yields of CAR T cells produced from elutriated lymphocytes with the yields of CAR T cells previous produced from cells isolated from PBMC concentrates by anti-CD3/CD28 bead selection or by anti-CD3/CD28 bead selection plus plastic adherence found that greater quantities of GD2-CAR T cells were produced from elutriated lymphocytes, but not CD19-CAR T cells.
  
  
  
  Enrichment of PBMC concentrates for lymphocytes using elutriation increased the quantity of GD2-CAR T cells produced. These results provide further evidence that CAR T cell expansion is inhibited by monocytes and granulocytes.
authors.affiliation: Cell Processing Section, Department of Transfusion Medicine, NIH Clinical Center, NIH, 10 Center Drive-MSC-1184, Building 10, Room 3C720, Bethesda, MD, 20892-1184, USA. dstroncek@cc.nih.gov.
authors.forename: David F
authors.identifier: 0000-0001-5867-3265
authors.initials: DF
authors.lastname: Stroncek
authors.affiliation: Division of Pediatric Hematology/Oncology, Department of Pediatrics, University of Virginia, Charlottesville, USA.
authors.forename: Daniel W
authors.initials: DW
authors.lastname: Lee
authors.affiliation: Cell Processing Section, Department of Transfusion Medicine, NIH Clinical Center, NIH, 10 Center Drive-MSC-1184, Building 10, Room 3C720, Bethesda, MD, 20892-1184, USA.
authors.forename: Jiaqiang
authors.initials: J
authors.lastname: Ren
authors.affiliation: Cell Processing Section, Department of Transfusion Medicine, NIH Clinical Center, NIH, 10 Center Drive-MSC-1184, Building 10, Room 3C720, Bethesda, MD, 20892-1184, USA.
authors.forename: Marianna
authors.initials: M
authors.lastname: Sabatino
authors.affiliation: Cell Processing Section, Department of Transfusion Medicine, NIH Clinical Center, NIH, 10 Center Drive-MSC-1184, Building 10, Room 3C720, Bethesda, MD, 20892-1184, USA.
authors.forename: Steven
authors.initials: S
authors.lastname: Highfill
authors.affiliation: Cell Processing Section, Department of Transfusion Medicine, NIH Clinical Center, NIH, 10 Center Drive-MSC-1184, Building 10, Room 3C720, Bethesda, MD, 20892-1184, USA.
authors.forename: Hanh
authors.initials: H
authors.lastname: Khuu
authors.affiliation: Pediatric Oncology Branch, Center for Cancer Research, National Cancer Center, NIH, Bethesda, USA.
authors.forename: Nirali N
authors.initials: NN
authors.lastname: Shah
authors.affiliation: Pediatric Oncology Branch, Center for Cancer Research, National Cancer Center, NIH, Bethesda, USA.
authors.forename: Rosandra N
authors.initials: RN
authors.lastname: Kaplan
authors.affiliation: Pediatric Oncology Branch, Center for Cancer Research, National Cancer Center, NIH, Bethesda, USA.
authors.forename: Terry J
authors.initials: TJ
authors.lastname: Fry
authors.affiliation: Parker Institute for Cancer Immunotherapy, Stanford University, Stanford, USA.
authors.forename: Crystal L
authors.initials: CL
authors.lastname: Mackall
mesh_terms: D015703:Antigens, CD / Q000378:metabolism; D002448:Cell Adhesion; D002648:Child; D006801:Humans; D018655:Lymphocyte Count; D011948:Receptors, Antigen, T-Cell / Q000378:metabolism*; D013601:T-Lymphocytes / Q000276:immunology*; D014161:Transduction, Genetic; D055815:Young Adult
chemical_list: D015703:Antigens, CD; C000605956:CD19-specific chimeric antigen receptor; D011948:Receptors, Antigen, T-Cell
keywords: Cancer immunotherapy; Cellular therapy; Chimeric antigen receptor T cells; Elutriation; Myeloid derived suppressor cells; Peripheral blood mononuclear cells; T cells