title: GMP production and testing of Xcellerated T Cells for the treatment of patients with CLL.
abstract: BACKGROUND
  Pre-clinical studies suggest Xcellerated T Cells have the potential to produce a potent anti-tumor effect, restore broad immune function and reduce the risk of infectious complications in patients with CLL. Unlike other cancer settings, T cells constitute only a small fraction of CLL patients' PBMC. To generate large numbers of Xcellerated T Cells of high purity from CLL patients' PBMC, a reproducible, streamlined and cost-effective good manufacturing process (GMP) is required.
  
  
  METHODS
  The 10-L volume Wave Bioreactor-based Xcellerate III Process using Xcyte Dynabeads in a single custom 20-L Cellbag container was adapted, qualified and implemented for GMP operations.
  
  
  RESULTS
  For n=17 CLL patients, starting with approximately 1.34 x 10(9) CD3+ T cells at 6.8+/-7.5% purity in the PBMC leukapheresis products, using the 10-L volume Wave Bioreactor-based Xcellerate III Process, it was feasible to manufacture 137.0+/-34.3 x 10(9) Xcellerated T Cells at 98.5+/-1.0% CD3+ T-cell purity. An average 400-fold clearance of malignant B cells was documented during the manufacturing process. The Xcellerated T Cells produced from the Xcellerate III Process exhibited high in vitro biologic activity and have their T-cell receptor repertoire restored to a normal diversity. In-process T-cell activation was reproducibly robust, as measured by increase in cell size, up-regulation of CD25 and CD154 expression and the secretion of IL-2, IFN-gamma and tumor necrosis factor (TNF)-alpha.
  
  
  CONCLUSIONS
  A low-volume, high-yield bioreactor-based process has been developed, qualified and implemented for the reproducible, GMP manufacture of high purity, biologically active Xcellerated T Cells for the treatment of CLL patients in clinical trials.
authors.affiliation: Xcyte Therapies Inc. Seattle Washington 98104, USA.
authors.forename: L S
authors.initials: LS
authors.lastname: Hami
authors.forename: C
authors.initials: C
authors.lastname: Green
authors.forename: N
authors.initials: N
authors.lastname: Leshinsky
authors.forename: E
authors.initials: E
authors.lastname: Markham
authors.forename: K
authors.initials: K
authors.lastname: Miller
authors.forename: S
authors.initials: S
authors.lastname: Craig
mesh_terms: D001402:B-Lymphocytes / Q000166:cytology / Q000378:metabolism; D019149:Bioreactors; D002469:Cell Separation; D016207:Cytokines / Q000378:metabolism; D006801:Humans; D016219:Immunotherapy, Adoptive; D015451:Leukemia, Lymphocytic, Chronic, B-Cell / Q000628:therapy*; D008213:Lymphocyte Activation; D011948:Receptors, Antigen, T-Cell / Q000378:metabolism; D016176:T-Lymphocyte Subsets; D013601:T-Lymphocytes / Q000166:cytology* / Q000276:immunology* / Q000378:metabolism
chemical_list: D016207:Cytokines; D011948:Receptors, Antigen, T-Cell