Rationale for the selection of ricin A-chain anti-T immunotoxins for mature T cell depletion.

A series of 25 different ricin A-chain immunotoxins (IT) were prepared with monoclonal antibodies reacting with several T cell antigens belonging to different clusters of differentiation (CD) to select the most appropriate immunotoxins (IT) for mature T cell depletion. Our screening procedure was performed in 2 steps. First, IT were evaluated using protein synthesis inhibition assay on clonogenic malignant cells in order to determine the most active IT for a given CD. Second, IT thus selected were evaluated for both mature T cell killing efficacy and tolerance on hematopoietic progenitor cells (HPC). This study showed that (1) different IT directed against the same CD antigen displayed a wide range of activity, suggesting the need for a large screening of IT to determine the most appropriate antibody for a given target antigen; (2) anti-CD3 and anti-CD5 ricin A-chain IT are the most active, displaying a cytoreduction of more than 2 logs on mature T cells; (3) the 3 different anti-CD2 ricin A-chain IT evaluated in this study induced poor mature T cell cytoreduction despite their relative efficacy on leukemic cells; (4) anti-CD8 ricin A-chain IT showed almost no efficacy on relevant target cells; (5) no toxicity on HPC was found for concentrations up to 10(-8) of A-chain whatever the IT used.