Large-scale generation of autologous dendritic cells for immunotherapy in patients with acute myeloid leukemia.
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.
Schmitt
Anita
A
Institute for Clinical Transfusion Medicine and Immunogenetics, Department of Pathology, and the Clinic for Internal Medicine III, University of Ulm, Ulm, Germany.
Reinhardt
Peter
P
Hus
Iwona
I
Tabarkiewicz
Jacek
J
Roliñski
Jacek
J
Barth
Thomas
T
Giannopoulos
Krzysztof
K
Dmoszyñska
Anna
A
Wiesneth
Markus
M
Schmitt
Michael
M
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