title: Analysis of CD34 populations in mobilised peripheral blood stem cell harvests and in bone marrow by fluorescent in situ hybridisation for the bcr/abl gene fusion in patients with chronic granulocytic leukaemia.
abstract: For those patients ineligible for allogeneic bone marrow transplant and who are non-responsive to interferon, autotransplant with peripheral blood stem cells (PBSC) mobilised after intensive chemotherapy, may provide a novel approach to improve prognosis in patients with chronic granulocytic leukaemia. PBSC harvests are assessed for CD34-positive cell numbers, which serve as an indicator of engraftment potential, and are also analysed cytogenetically to ascertain tumour cell contamination. However, a more accurate assessment of PBSC harvest contamination requires investigation of the Philadelphia (Ph) status of the CD34pos population, in which the cells that provide long-term engraftment are contained. In this study, we have analysed these levels in mobilised PBSC and also in bone marrow (BM) harvests, taken several weeks prior to mobilising chemotherapy. Using fluorescent in situ hybridisation for the bcr/abl gene fusion, we have shown that the median number of Ph negative cells in CD34pos isolated populations was 14.95% in BM compared to 79.05% in PBSC harvests and that in all PBSC samples tested, Ph positivity in CD34pos populations was always detectable either by FISH or one round PCR methods. In paired assessments of both PBSC and BM harvests, higher levels of Ph negative CD34pos cells (> or = 14%) isolated from BM harvests, taken prior to intensive chemotherapy, correlated with higher levels of Ph negative CD34pos cells (> or = 78.5%) in PBSC harvests. These data may aid in the selection of patients for whom PBSC harvesting, after mobilisation, is more likely to achieve an autograft product containing predominantly Ph negative CD34pos cells and may exclude those patients for whom the risk, morbidity and expense of stem cell harvesting may have no apparent benefit over a chronic phase BM harvest.
authors.affiliation: Department of Haematology, Royal Victoria Infirmary, Newcastle upon Tyne, UK.
authors.forename: J A
authors.initials: JA
authors.lastname: Irving
authors.forename: A
authors.initials: A
authors.lastname: Lennard
authors.forename: N
authors.initials: N
authors.lastname: Storey
authors.forename: J
authors.initials: J
authors.lastname: Conn
authors.forename: J
authors.initials: J
authors.lastname: Dunn
authors.forename: K
authors.initials: K
authors.lastname: Oliver
authors.forename: S J
authors.initials: SJ
authors.lastname: Proctor
authors.forename: A M
authors.initials: AM
authors.lastname: Dickinson
mesh_terms: D000328:Adult; D018952:Antigens, CD34 / Q000378:metabolism*; D001854:Bone Marrow Cells / Q000276:immunology; D005260:Female; D016044:Fusion Proteins, bcr-abl / Q000235:genetics*; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000097:blood / Q000235:genetics* / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D010677:Philadelphia Chromosome; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013234:Stem Cells / Q000276:immunology
chemical_list: D018952:Antigens, CD34; D016044:Fusion Proteins, bcr-abl