A negative-selection strategy for depleting myeloma cells from patients' BM and/or leukapheresis blood

Abstract

Autologous transplantation improves survival in multiple myeloma patients, however, most eventually relapse. As an attempt towards improving relapse-free survival, we designed a negative-selection purging strategy, to remove myeloma cells from leukapheresis harvests using MAbs specific for Ags on myeloma cells. CD38 is highly expressed on myeloma plasma cells, but expressed at lower levels on normal progenitors and absent on in vivo repopulating cells. We evaluated depletion of CD38-expressing cells, with or without depletion of B-cell Ag-expressing cells. Using myeloma BM or blood cells diluted into allogeneic G-CSF primed leukapheresis cells, bispecific tetrameric Ab complexes that bind dextran iron particles were used to label and retain cells in a magnetic column, StemSep. Depletion efficacy was measured by semi-quantitative allele-specific oligonucleotide (ASO)-PCR amplification of patients' clonotypic IgH gene. Low (0.2 microg/mL) concentrations of anti-CD38 with CD19 and CD20 complexes depleted approximately 3-5 logs of clonotypic cells, with recovery of approximately 19% of colony-forming cells, approximately 50% primitive progenitors measured by LTCIC and retention of non-obese diabetic /SCID engrafting ability. Scale-up experiments using leukapheresis harvests and 0.5-1 x 10(10) cell capacity columns demonstrated no loss of log depletion of highly positive cells, or recovery of unlabelled cells. These results compare favorably with other purging techniques and allow the retention of most normal BM cells, including T cells, which may be important for immunity. These results support the development of a clinical trial using this strategy for purging myeloma cells.