In vitro interleukin-3 binding to leukemia cells predicts cytotoxicity of a diphtheria toxin/IL-3 fusion protein.
Patients with acute myeloid leukemia frequently develop chemotherapy resistant blasts. To overcome multidrug resistance, a diphtheria toxin fusion protein (DTIL3) was engineered by fusing the catalytic and translocation domains of diphtheria toxin (DT) to human interleukin-3 (IL-3). However, when blasts were isolated from patients and tested for colony growth inhibition by DTIL3, only a third of the samples showed sensitivity to the fusion protein. Prior to clinical development, we need to be able to identify which patients are likely to respond to therapy with DTIL3. In this report, we compared the inhibition of thymidine incorporation in human leukemia cell lines by DTIL3 to the IL-3 receptor number and affinity. We found DTIL3 was cytotoxic to four of the eight cell lines tested with half-maximal inhibition of thymidine incorporation (IC(50)) from 1 to 50 pM. The IL-3 receptor density for these cell lines ranged from 0 to 2635 receptors per cell. The dissociation constant for an IL-3 high-affinity receptor agonist was 0.5 nM for cell lines with receptors. We found a correlation for the cell lines between the presence of high-affinity IL-3 receptors and sensitivity to DTIL3 (p = 0.03). These results suggest the variability in sensitivity of patient leukemic progenitors to DTIL3 may be due in part to the presence or absence of high-affinity IL-3 receptors.
Alexander
R L
RL
Department of Physiology and Pharmacology, Wake Forest University School of Medicine, Wake Forest, North Carolina 27157, USA.
Kucera
G L
GL
Klein
B
B
Frankel
A E
AE
D000970:Antineoplastic Agents / Q000494:pharmacology*; D002455:Cell Division / Q000187:drug effects*; D004167:Diphtheria Toxin / Q000494:pharmacology*; D006801:Humans; D007377:Interleukin-3 / Q000378:metabolism* / Q000494:pharmacology*; D007938:Leukemia / Q000378:metabolism* / Q000473:pathology; D011485:Protein Binding; D016185:Receptors, Interleukin-3 / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000494:pharmacology; D014407:Tumor Cells, Cultured
D000970:Antineoplastic Agents; D004167:Diphtheria Toxin; D007377:Interleukin-3; D016185:Receptors, Interleukin-3; D011993:Recombinant Fusion Proteins; C407154:diphtheria toxin-interleukin-3 fusion protein, recombinant