title: Dendritic cell-based vaccines in the setting of peripheral blood stem cell transplantation: CD34+ cell-depleted mobilized peripheral blood can serve as a source of potent dendritic cells.
abstract: We are investigating the use of tumor-pulsed dendritic cell (DC)-based vaccines in the treatment of patients with advanced cancer. In the current study, we evaluated the feasibility of obtaining both CD34+ hematopoietic stem/ progenitor cells (HSCs) and functional DCs from the same leukapheresis collection in adequate numbers for both peripheral blood stem cell transplantation (PBSCT) and immunization purposes, respectively. Leukapheresis collections of mobilized peripheral blood mononuclear cells (PBMCs) were obtained from normal donors receiving granulocyte colony-stimulating factor (G-CSF) (for allogeneic PBSCT) and from intermediate grade non-Hodgkin's lymphoma or multiple myeloma patients receiving cyclophosphamide plus G-CSF (for autologous PBSCT). High enrichment of CD34+ HSCs was obtained using an immunomagnetic bead cell separation device. After separation, the negative fraction of mobilized PBMCs from normal donors and cancer patients contained undetectable levels of CD34+ HSCs by flow cytometry. This fraction of cells was then subjected to plastic adherence, and the adherent cells were cultured for 7 days in GM-CSF (100 ng/ml) and interleukin 4 (50 ng/ml) followed by an additional 7 days in GM-CSF, interleukin 4, and tumor necrosis factor alpha (10 ng/ml) to generate DCs. Harvested DCs represented yields of 4.1+/-1.4 and 5.8+/-5.4% of the initial cells plated from the CD34+ cell-depleted mobilized PBMCs of normal donors and cancer patients, respectively, and displayed a high level expression of CD80, CD86, HLA-DR, and CD11c but not CD14. This phenotypic profile was similar to that of DCs derived from non-CD34+ cell-depleted mobilized PBMCs. DCs generated from CD34+ cell-depleted mobilized PBMCs elicited potent antitetanus as well as primary allogeneic T-cell proliferative responses in vitro, which were equivalent to DCs derived from non-CD34+ cell-depleted mobilized PBMCs. Collectively, these results demonstrate the feasibility of obtaining both DCs and CD34+ HSCs from the same leukapheresis collection from G-CSF-primed normal donors and cancer patients in sufficient numbers for the purpose of combined PBSCT and immunization strategies.
authors.affiliation: Department of Surgery, University of Michigan Medical Center, Ann Arbor 48109-0666, USA.
authors.forename: D
authors.initials: D
authors.lastname: Choi
authors.forename: M
authors.initials: M
authors.lastname: Perrin
authors.forename: S
authors.initials: S
authors.lastname: Hoffmann
authors.forename: A E
authors.initials: AE
authors.lastname: Chang
authors.forename: V
authors.initials: V
authors.lastname: Ratanatharathorn
authors.forename: J
authors.initials: J
authors.lastname: Uberti
authors.forename: K T
authors.initials: KT
authors.lastname: McDonagh
authors.forename: J J
authors.initials: JJ
authors.lastname: Mulé
mesh_terms: D018952:Antigens, CD34 / Q000276:immunology*; D019496:Cancer Vaccines / Q000627:therapeutic use; D002469:Cell Separation; D002478:Cells, Cultured; D003713:Dendritic Cells / Q000276:immunology* / Q000637:transplantation; D005240:Feasibility Studies; D005434:Flow Cytometry; D016179:Granulocyte Colony-Stimulating Factor / Q000502:physiology; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000276:immunology*; D006801:Humans; D007114:Immunization; D007937:Leukapheresis; D007963:Leukocytes, Mononuclear / Q000276:immunology; D016131:Lymphocyte Subsets / Q000276:immunology; D009369:Neoplasms / Q000276:immunology / Q000628:therapy; D010641:Phenotype; D019172:Transplantation Conditioning
chemical_list: D018952:Antigens, CD34; D019496:Cancer Vaccines; D016179:Granulocyte Colony-Stimulating Factor