title: Assessment of proliferative responses to granulocyte-macrophage colony-stimulating factor (GM-CSF) in acute myeloid leukaemia using a fluorescent ligand for the nucleoside transporter.
abstract: Nucleoside transporter expression has been linked to proliferation in a variety of haemopoietic cell types. Granulocyte-macrophage colony-stimulating factor (GM-CSF) was given for 72 h before commencing chemotherapy in 15 patients with relapsed or refractory acute myeloid leukaemia (AML) and in 11 patients serial bone marrows were taken for measurement of [3H]thymidine labelling index, Ki-67 positivity and maximal binding of 5-(SAENTA-x8)-fluorescein, a flow cytometry ligand which enumerates nucleoside transporter sites. GM-CSF caused proliferation of marrow myeloblasts in eight of 11 patients, while in three patients there was no change in proliferative indices. The expression of nucleoside transporters increased up to 4-fold in the myeloblasts from the patients showing a proliferative response to GM-CSF but there was no increase in transporters on the myeloblasts from the three non-responding patients. A close correlation was found between the fold increase in nucleoside transporter expression and the fold increase in labelling index of marrow myeloblasts (r = 0.86, n = 9, p < 0.01). In one patient with acute megakaryoblastic leukemia, GM-CSF caused parallel increases in labelling index, Ki-67 positivity and numbers of nucleoside transporters on peripheral blood blast cells. Thus induction of proliferation by cytokine increases the expression of nucleoside transporters on leukaemic myeloblasts studied in serial samples from the same source (bone marrow or blood). The suitability of 5-(SAENTA-x8)-fluorescein for two colour flow cytometric analysis allows the rapid enumeration of nucleoside transporters in the myeloblast compartment of heterogeneous marrow samples.
authors.affiliation: Department of Haematology, Austin Hospital, Victoria, Australia.
authors.forename: J S
authors.initials: JS
authors.lastname: Wiley
authors.forename: J S
authors.initials: JS
authors.lastname: Cebon
authors.forename: G P
authors.initials: GP
authors.lastname: Jamieson
authors.forename: J
authors.initials: J
authors.lastname: Szer
authors.forename: J
authors.initials: J
authors.lastname: Gibson
authors.forename: R K
authors.initials: RK
authors.lastname: Woodruff
authors.forename: J J
authors.initials: JJ
authors.lastname: McKendrick
authors.forename: W P
authors.initials: WP
authors.lastname: Sheridan
authors.forename: J C
authors.initials: JC
authors.lastname: Biggs
authors.forename: M B
authors.initials: MB
authors.lastname: Snook
mesh_terms: D000208:Acute Disease; D001798:Blood Proteins / Q000032:analysis; D001853:Bone Marrow / Q000187:drug effects; D001854:Bone Marrow Cells; D002352:Carrier Proteins / Q000032:analysis*; D002455:Cell Division / Q000187:drug effects; D005452:Fluoresceins; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000627:therapeutic use*; D006801:Humans; D007947:Leukemia, Megakaryoblastic, Acute / Q000188:drug therapy / Q000473:pathology; D007951:Leukemia, Myeloid / Q000188:drug therapy* / Q000473:pathology*; D008565:Membrane Proteins / Q000032:analysis*; D033703:Nucleoside Transport Proteins; D011684:Purine Nucleosides; D013936:Thymidine / Q000378:metabolism; D014316:Tritium
chemical_list: D001798:Blood Proteins; D002352:Carrier Proteins; D005452:Fluoresceins; D008565:Membrane Proteins; D033703:Nucleoside Transport Proteins; D011684:Purine Nucleosides; D014316:Tritium; C079645:5-(SAENTA-x8)fluorescein; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D013936:Thymidine