Table 4.

Stimulation of [3H]Thymidine Uptake by AML Cells in Response to Leptin

Patient Diagnosis Source Blasts, % rhL IL-3G-SCF SCF IL-3 + rhL G-CSF + rhL SCF + rhL Long Isoform Short Isoform
1  M5 BM 85  1.65 3.58  0.9  1.29 4.2 1.1  1.57  +  +  
2  M1 BM 87  0.67  1.02  0.85  4.2  4.6 1.02  4.12  ++  +  
3  M1  BM 88  0.65 0.43  0.95  0.7  0.65  0.9  0.7  +  ND  
4 M0  BM 81  0.8  1.62  0.61  0.78  1.73  0.53 0.94  +  ND  
5  M2  BM 88  1  73.1  1.2 23.07  85.9 3.9 29.5 +  ND  
6  M1  BM 94  0.96  1.28  1.32 8.05  1.6 1.02  7.19  +  ++  
7 M2  BM 88  1.2  3.1  1  9.4  3.28  1.23  9 ++  ++  
8  M2  PB 84  1.14  43.3  2.78 7.5  43.8  5.27 43.8 ++ ++  
9  Secondary AML  BM 82  1.8 24 2.4  22.3  25.5  0.85  20.4  +  +  
10 Secondary AML  BM 81  1.1  4.3  0.9  29.3  3.5 0.7  32.8 +  +  
11  M1, relapse PB 93  1.5 2.06  1.1  1.14  1.42 0.14  1.15  +  ++  
12  M1, relapse  BM 90 1.07  1.17  2.54  11.9  0.9  2.78  11.48  ++ ++  
13  M4, relapse  BM 90  1.3  4.3  1.15 6.2  3  1.7 6.4  +  +  
14  M1, relapse  BM 89  1.01  0.72  0.9  3.05  0.9  0.27 3.36  +  + 
Patient Diagnosis Source Blasts, % rhL IL-3G-SCF SCF IL-3 + rhL G-CSF + rhL SCF + rhL Long Isoform Short Isoform
1  M5 BM 85  1.65 3.58  0.9  1.29 4.2 1.1  1.57  +  +  
2  M1 BM 87  0.67  1.02  0.85  4.2  4.6 1.02  4.12  ++  +  
3  M1  BM 88  0.65 0.43  0.95  0.7  0.65  0.9  0.7  +  ND  
4 M0  BM 81  0.8  1.62  0.61  0.78  1.73  0.53 0.94  +  ND  
5  M2  BM 88  1  73.1  1.2 23.07  85.9 3.9 29.5 +  ND  
6  M1  BM 94  0.96  1.28  1.32 8.05  1.6 1.02  7.19  +  ++  
7 M2  BM 88  1.2  3.1  1  9.4  3.28  1.23  9 ++  ++  
8  M2  PB 84  1.14  43.3  2.78 7.5  43.8  5.27 43.8 ++ ++  
9  Secondary AML  BM 82  1.8 24 2.4  22.3  25.5  0.85  20.4  +  +  
10 Secondary AML  BM 81  1.1  4.3  0.9  29.3  3.5 0.7  32.8 +  +  
11  M1, relapse PB 93  1.5 2.06  1.1  1.14  1.42 0.14  1.15  +  ++  
12  M1, relapse  BM 90 1.07  1.17  2.54  11.9  0.9  2.78  11.48  ++ ++  
13  M4, relapse  BM 90  1.3  4.3  1.15 6.2  3  1.7 6.4  +  +  
14  M1, relapse  BM 89  1.01  0.72  0.9  3.05  0.9  0.27 3.36  +  + 

Mononuclear cells from patient samples were cultured in triplicate in the presence or absence of recombinant human leptin (100 ng/mL) in 96-well flat-bottom microtiter well plates (2.5 × 104cells/0.2 mL/well). For costimulation experiments, rHG-CSF was added to cultures at concentrations of 100 U/mL; IL-3 and SCF were added at 50 and 100 ng/mL, respectively. After 2 days of incubation, 0.1 μCi of [3H]thymidine was added to each well, and the cultures were continued overnight. The cells were then deposited onto harvester filters, and radioactivity was determined in a scintillation counter. The stimulation index was calculated as the ratio of [3H]thymidine incorporated in leptin-treated and untreated samples in counts per minute. Underscoring indicatesP < .05.

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