title: Efficacy and safety of CD34-selected and CD19-depleted autografting in multiple myeloma patients: a pilot study.
abstract: OBJECTIVE
  If multiple myeloma patients are to be cured after high-dose treatment supported by autologous stem cell transplantation, grafts must be purged of circulating myeloma cells. Myeloma cells are present in all grafts and have been identified as CD38(++)CD45(-) plasma cells, plasma blasts, and CD19(+) B cells.
  
  
  METHODS
  In an attempt to improve the purging strategy, we studied a two-step procedure consisting of CD34(+) enrichment followed by CD19 depletion. This article describes the evaluation of this sequential magnetic microbead selection after 18 procedures in 14 patients.
  
  
  RESULTS
  The processed autografts contained a median CD34 purity of 81% (range 21-99%) and a recovery of 47% (range 15-82%). Flow cytometric analysis documented the expected reduction of CD34(-) B cells and plasma cells, in most cases to a level below the sensitivity of flow cytometry. Real-time reverse transcriptase polymerase chain reaction documented a CD19 mRNA relative reduction to 0.042 (range 0.01-0.21). Allele-specific oligonucleotide IgH primers were designed for five patients. All products were positive for clonal myeloma cells before processing, but only 1 of 5 was negative after the procedure. The clinical outcome after reinfusion of the processed autografts was evaluated by blood cell recovery and found to be within the range expected from engraftment of unmanipulated autografts. One patient who had delayed platelet recovery associated with cytomegalovirus infection recovered after anti-cytomegalovirus treatment.
  
  
  CONCLUSIONS
  This pilot study documented engraftment after reinfusion of CD34-selected and CD19-depleted autografts. However, one patient suffered from unexpected prolonged thrombocytopenia. The efficacy of the procedure was evaluated and reduction of myeloma cells was indicated, with only one autograft free of clonal cells.
authors.affiliation: The Stem Cell Laboratory, Department of Haematology, Herlev Hospital, University of Copenhagen, Herlev, Denmark.
authors.forename: Thomas
authors.initials: T
authors.lastname: Rasmussen
authors.forename: Bosse
authors.initials: B
authors.lastname: Björkstrand
authors.forename: Heidi
authors.initials: H
authors.lastname: Andersen
authors.forename: Eva
authors.initials: E
authors.lastname: Gaarsdal
authors.forename: Hans Erik
authors.initials: HE
authors.lastname: Johnsen
mesh_terms: D018941:Antigens, CD19; D018952:Antigens, CD34; D006085:Graft Survival; D019650:Hematopoietic Stem Cell Mobilization; D018380:Hematopoietic Stem Cell Transplantation; D006801:Humans; D018189:Immunomagnetic Separation; D009101:Multiple Myeloma / Q000628:therapy*; D010865:Pilot Projects; D014182:Transplantation, Autologous
chemical_list: D018941:Antigens, CD19; D018952:Antigens, CD34