Abstract: Elucidating the intracellular fate(s) of targeted toxins is of fundamental importance for their optimal use as anticancer drugs, since the biochemical targets of their enzymatic activity reside in the cell cytoplasm, as in the case of the plant ribosome inactivating proteins (RIP) saporin, ricin and of bacterial toxins. In this paper, we compared the cell surface binding and cytotoxic properties of the model RIP ricin to an immunotoxin constructed with a monoclonal antibody directed against the human T-cell marker CD7 covalently linked to saporin (CD7-SAP). Our results indicate that, despite the fact that internalization takes place via an apparently common entry route leading to the Golgi complex, surprisingly, the addition of an endoplasmic reticulum retrieval C-terminal signal (KDEL) to CD7-SAP does not potentiate its cytotoxicity. In addition, while ricin toxicity is clearly reduced by Brefeldin A under conditions where this fungal metabolite causes Golgi stack disruption, we paradoxically observed a potentiating effect by Brefeldin A on CD7-SAP cytotoxicity suggesting that this inhibitor interferes with retrograde route(s) other than the well established Trans-Golgi Network-ER retrograde route.
Keywords: endocytosis; immunotoxin; ricin; Brefeldin A; leukaemia cells
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Giansanti, F.; Giordani, V.; Vago, R.; Flavell, D.J.; Flavell, S.U.; Fabbrini, M.S.; Ippoliti, R. Dissecting the Entry Route of Saporin-based a-CD7 Immunotoxins in Human T-Cell Acute Lymphoblastic Leukaemia Cells. Antibodies 2013, 2, 50-65.
Giansanti F, Giordani V, Vago R, Flavell DJ, Flavell SU, Fabbrini MS, Ippoliti R. Dissecting the Entry Route of Saporin-based a-CD7 Immunotoxins in Human T-Cell Acute Lymphoblastic Leukaemia Cells. Antibodies. 2013; 2(1):50-65.
Giansanti, Francesco; Giordani, Valeria; Vago, Riccardo; Flavell, David J.; Flavell, Sopsamorn U.; Fabbrini, Maria S.; Ippoliti, Rodolfo. 2013. "Dissecting the Entry Route of Saporin-based a-CD7 Immunotoxins in Human T-Cell Acute Lymphoblastic Leukaemia Cells." Antibodies 2, no. 1: 50-65.