IL2 fused to lectin-deficient ricin is toxic to human leukemia cells expressing the IL2 receptor.
Interleukin-2 (IL2) fused to ricin B chain (RTB) with modifications of amino acid residues in each of three galactose-binding subdomains (1alpha, 1beta and 2gamma) was expressed in insect cells, purified by immunoaffinity chromatography and reassociated with ricin A chain (RTA). The fusion toxin-bound human leukemic cells with IL2 receptors and the binding was competed with IL2 but not asialofetuin. In contrast, binding was not observed with receptor negative human cell lines, and the fusion molecule very weakly bound asialofetuin (Kd= 10(-6)M), indicating lectin-deficient RTB. The IL2-lectin-deficient RTB-RTA intoxicated IL2 receptor bearing cells as well as ricin or IL2-wild-type RTB-RTA. While ricin and IL2-wild-type RTB-RTA were equally toxic to receptor negative cell lines, the IL2-lectin-deficient RTB-RTA was two-two and one half logs less cytotoxic to these cell lines. The sensitivity of receptor-positive cells to the lectin-deficient fusion protein suggests that high avidity intracellular galactose binding may not be required for ricin intoxication, at least in the case of IL2 receptor-targeted molecules. Furthermore, the potent selective cytotoxicity of the fusion protein suggests that the IL2-lectin-deficient RTB-RTA and similar ricin fusion molecules directed against other leukemic cell surface receptors provide a novel class of fusion toxins for therapy of human leukemias.
Frankel
A E
AE
Department of Medicine, Medical University of South Carolina, Charleston, USA.
Fu
T
T
Burbage
C
C
Chandler
J
J
Willingham
M C
MC
Tagge
E P
EP
D000911:Antibodies, Monoclonal / Q000378:metabolism; D006801:Humans; D000922:Immunotoxins / Q000737:chemistry / Q000378:metabolism*; D007376:Interleukin-2 / Q000378:metabolism*; D037102:Lectins; D015458:Leukemia, T-Cell / Q000378:metabolism*; D015375:Receptors, Interleukin-2 / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D012276:Ricin / Q000737:chemistry / Q000378:metabolism*; D014407:Tumor Cells, Cultured / Q000378:metabolism
D000911:Antibodies, Monoclonal; D000922:Immunotoxins; D007376:Interleukin-2; D037102:Lectins; D015375:Receptors, Interleukin-2; D011993:Recombinant Fusion Proteins; D012276:Ricin