Table 1.
Patient characteristics of CD25-positive AML.
Fig 1.
Gating strategy and flow-cytometric analysis of AML cells.
A, Cells from AML01 were analyzed by flow cytometry. After discrimination of doublet and dead cells, the cells were gated as Lin–CD45low cells, and then CD34+ cells were analyzed for CD38 and CD25 expression. B, CD38 and CD25 expressions on Lin–CD45lowCD34+ population in nine AML patients are shown as contour plots. Percentages of each cell fraction are indicated in the plot areas.
Fig 2.
Reconstitution of human leukemia with distinct expression of CD25 at the primary and secondary transplantations in a PDX model.
Lin–CD34+CD38– cells from AML01 (A), 02 (B), and 03 (C) were sorted into CD25-positive and -negative populations, and then transplanted to primary recipients. Expression of CD25 on engrafted Lin–CD34+ cells was analyzed. Engrafted BM cells from AML01 and 02 were sorted again into CD25-positive and -negative populations, and then transplanted into secondary recipients. Expression of CD25 and CD38 on engrafted Lin–CD34+ cells in secondary recipients was analyzed.
Table 2.
Results of PDX assays of CD25-positive AML.
Fig 3.
Reconstitution of human leukemia with distinct expression of CD25 at the primary transplantation into mice, with failure at the secondary transplantation.
Lin–CD34+CD38– cells from AML04 (A), 05 (B), and 06 (C) were sorted into CD25-positive and -negative populations, and then transplanted to primary recipients. Expression of CD3/CD19, CD33, CD34, CD38, and CD25 on engrafted cells was analyzed. The hCD45+ cells were sorted and transplanted to secondary recipients. No engraftment was observed in mice transplanted with hCD45+ cells from AML04, 05, or 06.
Fig 4.
Cell culture of CD25-positive and -negative CD34+ cells from CD25-positive AML.
CD25-positive and -negative CD34+ cells from AML01 and 05 were isolated and cultured for 48 hours at a concentration of 3 × 105 /ml in 12-well plates in the presence of IL-3, G-CSF, GM-CSF, EPO, TPO, and SCF.