title: Large-scale generation of autologous dendritic cells for immunotherapy in patients with acute myeloid leukemia.
abstract: BACKGROUND
  Mononuclear cells (MNCs) of severely impaired acute myeloid leukemia (AML) patients may be collected by leukapheresis for large-scale generation of dendritic cells (AML-DCs) under good manufacturing practice (GMP) conditions for adoptive immunotherapy.
  
  
  METHODS
  In five end-stage AML patients, a leukapheresis procedure was performed with a cell separator (either COBE Spectra [Gambro BCT] or Amicus [Baxter]). For large-scale AML-DC generation, the MNCs of a single leukapheresis concentrate were isolated by density gradient and plated into a cell factory under GMP conditions. The AML-DCs were harvested on Day 8 of culture, and their viability, the mature morphology, and the phenotype were evaluated. The AML-DCs were injected subcutaneously into five AML patients up to four times at a biweekly interval.
  
  
  RESULTS
  All AML patients entered the leukapheresis procedure with a highly pathologic blood count. In a mean separation time of 198 +/- 33 minutes, a mean of 1.3 +/- 0.2-fold the total blood volume was processed with a white blood cell (WBC) yield of 9 x 10(9) to 70 x 10(9) per collection dependent on the precollection WBC count. After density gradient a mean of 2.2 x 10(9) +/- 0.3 x 10(9) MNCs were plated into a cell factory. This resulted in a mean viable and mature DC yield of 0.01 x 10(9) of MNCs.
  
  
  CONCLUSIONS
  The leukapheresis procedure is a feasible and safe procedure even in patients with hematologic malignancies and highly pathologic blood counts. Sufficient amounts of MNCs can be collected in leukopenic patients and the large-scale generation of AML-DCs in cell factories under GMP conditions yields in an adequate quantity of viable and mature AML-DCs.
authors.affiliation: Institute for Clinical Transfusion Medicine and Immunogenetics, Department of Pathology, and the Clinic for Internal Medicine III, University of Ulm, Ulm, Germany.
authors.forename: Anita
authors.initials: A
authors.lastname: Schmitt
authors.forename: Peter
authors.initials: P
authors.lastname: Reinhardt
authors.forename: Iwona
authors.initials: I
authors.lastname: Hus
authors.forename: Jacek
authors.initials: J
authors.lastname: Tabarkiewicz
authors.forename: Jacek
authors.initials: J
authors.lastname: Roliñski
authors.forename: Thomas
authors.initials: T
authors.lastname: Barth
authors.forename: Krzysztof
authors.initials: K
authors.lastname: Giannopoulos
authors.forename: Anna
authors.initials: A
authors.lastname: Dmoszyñska
authors.forename: Markus
authors.initials: M
authors.lastname: Wiesneth
authors.forename: Michael
authors.initials: M
authors.lastname: Schmitt
mesh_terms: D000368:Aged; D001804:Blood Transfusion, Autologous; D003713:Dendritic Cells / Q000637:transplantation*; D005260:Female; D006801:Humans; D016130:Immunophenotyping; D007167:Immunotherapy; D007937:Leukapheresis; D015470:Leukemia, Myeloid, Acute / Q000276:immunology / Q000473:pathology / Q000601:surgery* / Q000628:therapy*; D008297:Male; D008875:Middle Aged