title: Autologous lymphapheresis for the production of chimeric antigen receptor T cells.
abstract: The first step in manufacturing chimeric antigen receptor (CAR) T cells is to collect autologous CD3+ lymphocytes by apheresis. Patients, however, often have leukopenia or have other disease-related complications. We evaluated the feasibility of collecting adequate numbers of CD3+ cells, risk factors for inadequate collections, and the rate of adverse events.
  
  
  
  Apheresis lymphocyte collections from patients participating in three CAR T-cell clinical trials were reviewed. Collections were performed on the COBE Spectra by experienced nurses, with the goal of obtaining a minimum of 0.6 × 109and a target of 2 × 109CD3+ cells. Preapheresis peripheral blood counts, apheresis parameters, and product cell counts were analyzed.
  
  
  
  Of the 71 collections, 69 (97%) achieved the minimum and 55 (77%) achieved the target. Before apheresis, the 16 patients with yields below the target had significantly lower proportions and absolute numbers of circulating lymphocytes and CD3+ lymphocytes and higher proportions of circulating blasts and NK cells than those who achieved the target (470 × 106lymphocytes/L vs. 1340 × 106lymphocytes/L, p = 0.008; 349 × 106CD3+ cells/L vs. 914 × 106CD3+ cells/L, p = 0.001; 17.6% blasts vs. 4.55% blasts, p = 0.029). Enrichment of blasts in the product compared to the peripheral blood occurred in four patients, including the two patients whose collections did not yield the minimum number of CD3+ cells. Apheresis complications occurred in 11 patients (15%) and, with one exception, were easily managed in the apheresis clinic.
  
  
  
  In most patients undergoing CAR T-cell therapy, leukapheresis is well tolerated, and adequate numbers of CD3+ lymphocytes are collected.
authors.affiliation: Department of Transfusion Medicine, NIH Clinical Center, Bethesda, Maryland.
authors.forename: Elizabeth S
authors.identifier: 0000-0002-1004-3851
authors.initials: ES
authors.lastname: Allen
authors.affiliation: Department of Transfusion Medicine, NIH Clinical Center, Bethesda, Maryland.
authors.forename: David F
authors.initials: DF
authors.lastname: Stroncek
authors.affiliation: Department of Transfusion Medicine, NIH Clinical Center, Bethesda, Maryland.
authors.forename: Jiaqiang
authors.initials: J
authors.lastname: Ren
authors.affiliation: Department of Transfusion Medicine, NIH Clinical Center, Bethesda, Maryland.
authors.forename: Anne F
authors.initials: AF
authors.lastname: Eder
authors.affiliation: Department of Transfusion Medicine, NIH Clinical Center, Bethesda, Maryland.
authors.forename: Kamille A
authors.initials: KA
authors.lastname: West
authors.affiliation: Pediatric Oncology Branch, National Cancer Institute, National Institutes of Health, Bethesda, Maryland.
authors.forename: Terry J
authors.initials: TJ
authors.lastname: Fry
authors.affiliation: Division of Pediatric Hematology/Oncology, Department of Pediatrics, University of Virginia, Charlottesville, Virginia.
authors.forename: Daniel W
authors.initials: DW
authors.lastname: Lee
authors.affiliation: Pediatric Oncology Branch, National Cancer Institute, National Institutes of Health, Bethesda, Maryland.
authors.forename: Crystal L
authors.initials: CL
authors.lastname: Mackall
authors.affiliation: Department of Transfusion Medicine, NIH Clinical Center, Bethesda, Maryland.
authors.forename: Cathy
authors.initials: C
authors.lastname: Conry-Cantilena
mesh_terms: D000293:Adolescent; D000328:Adult; D017252:CD3 Complex / Q000032:analysis; D060846:Cell Engineering / Q000379:methods*; D002648:Child; D002675:Child, Preschool; D005260:Female; D006801:Humans; D007937:Leukapheresis / Q000379:methods* / Q000592:standards; D017710:Lymphocyte Transfusion / Q000009:adverse effects / Q000379:methods* / Q000592:standards; D008297:Male; D015202:Protein Engineering / Q000379:methods; D011948:Receptors, Antigen, T-Cell / Q000627:therapeutic use*; D014182:Transplantation, Autologous / Q000379:methods / Q000592:standards; D055815:Young Adult
chemical_list: D017252:CD3 Complex; D011948:Receptors, Antigen, T-Cell