Internalization of the cytotoxic molecules of T101 F(ab')2-(ricin-A-chain) immunotoxin into human T-leukemic cells.
We have investigated the internalization step of an immunotoxin and its relationship with cytotoxicity, with the F(ab')2-T101(ricin-A-chain) immunotoxin, directed against the CD5 antigen expressed on leukemic CEM cells. We first demonstrated that the biological action of the conjugate was related to its entry into the cell by an energy-dependent endocytotic process. We also found that during the first hours of cell intoxication, internalization is not the rate-limiting step of immunotoxin cytotoxicity. Internalization becomes limiting in cell intoxication only when the entry rate is low. Lastly we show that ammonium chloride, which strongly enhances immunotoxin potency, acts on internalized molecules for a very short time, suggesting that this enhancer affects an early intracellular step.
Department of Immunology, Sanofi Recherche, Montpellier, France.
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D000643:Ammonium Chloride / Q000494:pharmacology; D001692:Biological Transport; D002460:Cell Line; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000378:metabolism; D000922:Immunotoxins / Q000378:metabolism* / Q000494:pharmacology; D007700:Kinetics; D007938:Leukemia; D008985:Monensin / Q000494:pharmacology; D009363:Neoplasm Proteins / Q000096:biosynthesis; D012276:Ricin / Q000378:metabolism* / Q000494:pharmacology; D014407:Tumor Cells, Cultured / Q000378:metabolism
D007140:Immunoglobulin Fab Fragments; D000922:Immunotoxins; D009363:Neoplasm Proteins; D000643:Ammonium Chloride; D012276:Ricin; D008985:Monensin