Graft engineering of G-CSF-mobilized allogeneic leukapheresis products by counterflow centrifugal elutriation after CD34 column adsorption.
A major hindrance to the use of PBSC in allogeneic transplantation is the high rate of contamination with T lymphocytes, resulting in a considerable risk of GvHD. Natural killer (NK) cells are active against tumor cells but do not contribute to the development of GvHD. After adsorption of CD34+ cells of mobilized allogeneic leukapheresis products on a Ceprate column, we studied the separation of CD34 unadsorbed cells by counterflow centrifugal elutriation (CCE). Up to 1.0 x 10(10) cells were clearly separated into lymphocytes (fractions 110 and 140 ml/min), monocytes, and polymorphonuclear cells (fraction rotor off). Characterized by flow cytometry, T cells were distributed nearly equal to fractions 110 and 140. NK cells were concentrated 3.4-fold in fraction 140 as compared with the unseparated cells. The ratio of NK cells/T cells was improved by 33%. These results indicate that CCE is an effective method to enrich NK cells and to reduce T cells in stem cell separation products. Therefore, it is an option for adoptive therapy of cancer patients after transplantations (e.g., CML in relapse).
Bone Marrow Transplantation Center, University Hospital Eppendorf, Hamburg, Germany.
D
D
Skiera
W
W
Zeller
A R
AR
Zander
D018952:Antigens, CD34 / Q000276:immunology*; D000943:Antigens, Differentiation / Q000032:analysis; D018914:Cells, Immobilized / Q000737:chemistry; D003114:Colony-Forming Units Assay; D016238:Cytapheresis / Q000379:methods; D005434:Flow Cytometry; D016179:Granulocyte Colony-Stimulating Factor / Q000494:pharmacology*; D006801:Humans; D007937:Leukapheresis / Q000379:methods; D007958:Leukocyte Count; D007962:Leukocytes / Q000166:cytology; D008214:Lymphocytes / Q000276:immunology; D011994:Recombinant Proteins
D018952:Antigens, CD34; D000943:Antigens, Differentiation; D011994:Recombinant Proteins; D016179:Granulocyte Colony-Stimulating Factor