Characterization and receptor specific toxicity of two diphtheria toxin-related interleukin-3 fusion proteins DAB389-mIL-3 and DAB389-(Gly4Ser)2-mIL-3.
We have constructed two fusion proteins, DAB389-mIL-3 and DAB389-(Gly4Ser)2-mIL-3, in which the receptor-binding domain of diphtheria toxin is replaced by mouse interleukin-3 (IL-3). Cytotoxic activity of the fusion toxins was observed on three out of six cell lines assayed. This toxicity was mediated through binding to the IL-3 receptor as it was inhibited in a dose-dependent manner with murine IL-3 or anti-IL-3 neutralizing antibodies. DAB389-(Gly4Ser)2-mIL-3 was up to 5 times more toxic than DAB389-mIL-3, depending on the cell line (0.8 x 10(-10) M < IC50 < 3 x 10(-10) M). These proteins can be used for the detection of IL-3 receptors on mouse cells and should allow for the selective elimination of IL-3 receptor-positive pluripotent hematopoietic stem cells prior to bone marrow transplantation.
Liger
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Département d'Ingénierie et d'Etudes des Protéines (DIEP), DSV, CEA,CE Saclay, Gif-sur-Yvette, France.
vanderSpek
J C
JC
Gaillard
C
C
Cansier
C
C
Murphy
J R
JR
Leboulch
P
P
Gillet
D
D
D000818:Animals; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D004167:Diphtheria Toxin / Q000737:chemistry* / Q000378:metabolism / Q000633:toxicity; D007376:Interleukin-2 / Q000737:chemistry* / Q000378:metabolism / Q000633:toxicity; D007377:Interleukin-3 / Q000737:chemistry* / Q000378:metabolism / Q000633:toxicity; D051379:Mice; D017510:Protein Folding; D016185:Receptors, Interleukin-3 / Q000187:drug effects* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism / Q000633:toxicity
C105544:DAB(389)-(Gly(4)-Ser)2-interleukin 3; D004167:Diphtheria Toxin; D007376:Interleukin-2; D007377:Interleukin-3; D016185:Receptors, Interleukin-3; D011993:Recombinant Fusion Proteins; C078456:denileukin diftitox