title: Ex vivo graft purging and expansion of autologous blood progenitor cell products from patients with multiple myeloma.
abstract: Autologous peripheral blood progenitor cell (PBPC) transplantation is the treatment of choice for selected myeloma patients. However, tumor cells contaminating the apheresis product are a potential source of relapse. Here we report a sequential purging strategy targeting mature and immature clonogenic myeloma cell populations in the autograft. Thawed PBPC products of myeloma patients were treated with rituximab to kill CD138(-)20(+) B cells (highly clonogenic immature cells), and bortezomib to target CD138(+) cells (normal and differentiated myeloma plasma cells), followed by coculture with allogeneic mesenchymal stem cells (MSC) from normal donors. After 7 days of coculture, nonadherent cells were removed and cultured in the absence of MSC for an additional 7 days. Then, efficacy of purging (removal of CD138(-)20(+) and CD138(+) cells) was assessed by flow cytometry and PCR. We used our ex vivo purging strategy to treat frozen aphereses from 16 patients. CD138(+) and CD138(-)20(+)(19(+)) cells present in the initial products were depleted more than 3 and 4 logs, respectively based on 10(6) flow-acquisition events, and to levels below the limit of detection by PCR. In contrast, total nucleated cell (TNC), CD34(+) cell, and colony-forming cell numbers were increased by approximately 12 to 20, 8-, and 23-fold, respectively. Overall, ex vivo treatment of apheresis products with rituximab, bortezomib, and coculture with normal donor MSC depleted mature and immature myeloma cells from clinical aphereses while expanding the normal hematopoietic progenitor cell compartment.
authors.affiliation: Department of Stem Cell Transplantation & Cellular Therapy, University of Texas MD Anderson Cancer Center, Houston, TX 77030-4009, USA. hyang@mdanderson.org
authors.forename: Hong
authors.initials: H
authors.lastname: Yang
authors.forename: Simon N
authors.initials: SN
authors.lastname: Robinson
authors.forename: Yago
authors.initials: Y
authors.lastname: Nieto
authors.forename: Richard J
authors.initials: RJ
authors.lastname: Jones
authors.forename: Christopher D
authors.initials: CD
authors.lastname: Gocke
authors.forename: Junjun
authors.initials: J
authors.lastname: Lu
authors.forename: Sergio A
authors.initials: SA
authors.lastname: Giralt
authors.forename: Roy B
authors.initials: RB
authors.lastname: Jones
authors.forename: William K
authors.initials: WK
authors.lastname: Decker
authors.forename: Dongxia
authors.initials: D
authors.lastname: Xing
authors.forename: David
authors.initials: D
authors.lastname: Steiner
authors.forename: Richard E
authors.initials: RE
authors.lastname: Champlin
authors.forename: John D
authors.initials: JD
authors.lastname: McMannis
authors.forename: Jingjing
authors.initials: J
authors.lastname: Ng
authors.forename: Michael W
authors.initials: MW
authors.lastname: Thomas
authors.forename: Nina
authors.initials: N
authors.lastname: Shah
authors.forename: Borje S
authors.initials: BS
authors.lastname: Andersson
authors.forename: Simrit
authors.initials: S
authors.lastname: Parmar
authors.forename: Elizabeth J
authors.initials: EJ
authors.lastname: Shpall
mesh_terms: D058846:Antibodies, Monoclonal, Murine-Derived / Q000008:administration & dosage / Q000494:pharmacology*; D018952:Antigens, CD34 / Q000096:biosynthesis; D000971:Antineoplastic Combined Chemotherapy Protocols / Q000494:pharmacology; D016465:Bone Marrow Purging / Q000379:methods*; D001897:Boronic Acids / Q000008:administration & dosage / Q000494:pharmacology*; D000069286:Bortezomib; D045744:Cell Line, Tumor; D002469:Cell Separation / Q000379:methods; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000276:immunology; D006801:Humans; D008280:Magnetics; D009101:Multiple Myeloma / Q000188:drug therapy / Q000473:pathology / Q000601:surgery*; D036102:Peripheral Blood Stem Cell Transplantation / Q000379:methods*; D011719:Pyrazines / Q000008:administration & dosage / Q000494:pharmacology*; D000069283:Rituximab; D014182:Transplantation, Autologous
chemical_list: D058846:Antibodies, Monoclonal, Murine-Derived; D018952:Antigens, CD34; D001897:Boronic Acids; D011719:Pyrazines; D000069283:Rituximab; D000069286:Bortezomib