title: Polymerase chain reaction: a method for monitoring tumor cell purge by long-term culture in BCR/ABL positive acute lymphoblastic leukemia.
abstract: We report the successful purging of leukemia cells bearing the Philadelphia chromosome and BCR/ABL transcripts by long-term marrow culture (LTC), and subsequent grafting of the purged marrow in a case of refractory acute lymphoblastic leukemia. The efficiency of the purge was evaluated by polymerase chain reaction (PCR) for BCR/ABL transcripts. In two LTCs initiated in the blastic stage, we demonstrated the selective effect of three culture media (serum dependent, serum-free (SF) supplemented or not with IL3 and GM-CSF) on the proliferative potential of normal hematopoietic (CFU-GM/BFU-E) and leukemic progenitors (CFU-ALL). BCR/ABL positive cells disappeared after 3 to 4 weeks of culture. The addition of IL3 and GM-CSF to the SF medium enhanced the growth of CFU-GM/BFU-E and shortened the purging period. We therefore carried out a LTC in the presence of IL3 and GM-CSF with marrow harvested in morphological remission. BCR/ABL positivity was detected at the outset, although no leukemia cells could be identified. The BCR/ABL was no longer found by PCR in the 7 and 14 day LTCs. The patient, consolidated by high dose polychemotherapy and total body irradiation, was infused with the 14 day LTC. This study indicates that PCR is a useful and sensitive technique for monitoring tumor cell reduction after LTC prior to autografting.
authors.affiliation: Formation associée Claude Bernard sur les greffes de cellules souches hématopoïetiques, CHU Saint Antonie, Paris, France.
authors.forename: S
authors.initials: S
authors.lastname: Fabrega
authors.forename: J P
authors.initials: JP
authors.lastname: Laporte
authors.forename: M C
authors.initials: MC
authors.lastname: Giarratana
authors.forename: L
authors.initials: L
authors.lastname: Douay
authors.forename: L
authors.initials: L
authors.lastname: Fouillard
authors.forename: W M
authors.initials: WM
authors.lastname: Da
authors.forename: C
authors.initials: C
authors.lastname: Perrot
authors.forename: V
authors.initials: V
authors.lastname: Barbu
authors.forename: N C
authors.initials: NC
authors.lastname: Gorin
mesh_terms: D016465:Bone Marrow Purging; D016026:Bone Marrow Transplantation; D016895:Culture Media, Serum-Free; D016044:Fusion Proteins, bcr-abl / Q000032:analysis* / Q000235:genetics; D009363:Neoplasm Proteins / Q000032:analysis* / Q000235:genetics; D014411:Neoplastic Stem Cells / Q000737:chemistry*; D016133:Polymerase Chain Reaction; D015452:Precursor B-Cell Lymphoblastic Leukemia-Lymphoma / Q000235:genetics / Q000473:pathology / Q000601:surgery; D054198:Precursor Cell Lymphoblastic Leukemia-Lymphoma / Q000235:genetics / Q000473:pathology* / Q000601:surgery; D014182:Transplantation, Autologous
chemical_list: D016895:Culture Media, Serum-Free; D009363:Neoplasm Proteins; D016044:Fusion Proteins, bcr-abl