Clonality of cell populations in refractory anaemia using combined approach of gene loss and X-linked restriction fragment length polymorphism-methylation analyses.
We have used X-linked restriction fragment length polymorphism (RFLP)-methylation and gene deletion analyses to investigate the nature of the progenitor cell of origin in the myelodysplastic syndromes (MDS). Gene deletion studies were performed on the granulocyte and T-lymphocyte fractions of six women with refractory anaemia (RA) and either a partial deletion of the long arm of chromosome 5 (5q-) or monosomy 7. All six showed gene loss in the granulocyte but not the T-lymphocyte fractions, indicating monoclonality of the granulocytes but not the T-lymphocytes. In order to further investigate this finding, we subsequently performed X-RFLP-methylation studies using the probe M27 beta, and also a probe for the phosphoglycerate kinase (PGK) gene. These studies have confirmed the monoclonality of the granulocytes and the polyclonality of the T-lymphocytes in these cases. Our findings suggest that in this group of patients with MDS the T-lymphocytes were not involved in the disorder, and furthermore, in the one case where B-lymphocytes were also available, that the progenitor cell of origin was restricted to the myeloid lineage.
Leukaemia Research Fund Molecular and Cytogenetic Haematology Unit, John Radcliffe Hospital, Oxford.
G
G
Abrahamson
J
J
Boultwood
J
J
Madden
S
S
Kelly
D G
DG
Oscier
K
K
Rack
V J
VJ
Buckle
J S
JS
Wainscoat
D000753:Anemia, Refractory / Q000097:blood* / Q000235:genetics; D002872:Chromosome Deletion; D002895:Chromosomes, Human, Pair 5; D002897:Chromosomes, Human, Pair 7; D002999:Clone Cells / Q000473:pathology*; D015342:DNA Probes; D005260:Female; D008040:Genetic Linkage; D006098:Granulocytes / Q000473:pathology*; D006801:Humans; D009006:Monosomy; D010735:Phosphoglycerate Kinase / Q000235:genetics; D012150:Polymorphism, Restriction Fragment Length; D013601:T-Lymphocytes / Q000473:pathology*; D014960:X Chromosome
D015342:DNA Probes; D010735:Phosphoglycerate Kinase