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Blood, Vol. 113, Issue 24, 6206-6214, June 11, 2009
Chemosensitization of acute myeloid leukemia (AML) following mobilization by the CXCR4 antagonist AMD3100
Blood Nervi et al.
113: 6206
Supplemental materials for: Nervi et al
Files in this Data Supplement:
- Figure S1. Increased in vitro chemosensitivity of circulating APL cells (JPG, 74.6 KB)
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(A) Chemoensitivity of unpurified APLluc cells. Syngeneic B6129F1 recipient mice (n = 4) were intravenously injected with 106 APLluc cells. Two weeks after APLluc injection, cells were harvested from the peripheral blood, spleen and BM and incubated in the absence (control) or presence of 25 or 50 ng/mL AraC for 24 hours. APLluc cell survival was analyzed by bioluminescent imaging and results are presented as percent of the untreated control. Each bar represents the mean ± SD of a single experiment, for which each group was assayed in quadruplicate. (B) Chemosensitivity of purified APLluc cells. Syngeneic B6129F1 recipient mice (n = 4) were intravenously injected with 106 APLluc cells. Two weeks after APLluc injection, cells were harvested from the peripheral blood, spleen and BM and APLluc cells were purified to greater than 95% by CD34 immunoselection using an AutoMACS cell separation device (Miltenyi Biotech, Auburn, CA). The purity of isolated cells was quantified by flow cytometry. Purified APLluc cells were then incubated in the absence (control) or presence of 50 ng/mL AraC for 24 hours. APLluc cell survival was analyzed by bioluminescent imaging and results are presented as percent of the untreated control. Each bar represents the mean ± SD of a single experiment, for which each group was assayed in quadruplicate. * P < 0.05, ** P < 0.01, *** P < 0.001.
- Figure S2. AMD3100 sensitization of APL to Ara-C (JPG, 73.4 KB)
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Syngeneic B6129F1 recipient mice (n = 29) were intravenously injected with 106 AMLluc cells. Twelve days after AML injection, mice were left untreated (control; n = 6) or treated with AMD3100 alone (n = 7), Ara-C alone (n = 8), or the combination of AMD3100 and Ara-C (n = 8). Mice treated with chemotherapy received a single subcutaneous injection of Ara-C (500 mg/kg) on days 12 and 13 after AML injection. Mice treated with AMD3100 received subcutaneous injections of AMD3100 (5mg/kg) 1 hour before and 3 hours after each Ara-C injection. Total white blood cell counts (A) and AML blast cell counts (B) per microliter of peripheral blood in leukemic mice were determined by automated counting and flow cytometry of Gr1+CD34+ AML blast cells. The number of total WBCs and AMLluc cells in the peripheral blood at 19 and 23 days after injection were significantly reduced in mice treated with AMD3100 and Ara-C compared to mice treated with Ara-C alone. (C,D) Lack of general hematological toxicity in AMD3100 mobilized mice treated with chemotherapy. Mice treated with the combination of AMD3100 and Ara-C exhibit hemoglobin levels (C) and platelet counts (D) that are not significantly different than the Ara-C only treated control. * P < 0.05, ** P < 0.01. (skull and crossbones) indicates that no mice were alive in the AMD3100 only treatment group at 23 days after AML injection.
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