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The Journal of Immunology, Vol 152, Issue 5 2333-2343, Copyright © 1994 by American Association of Immunologists

ARTICLES

Sensitivity of human leukemia cells in exponential or stationary growth phase to anti-CD5 immunotoxins. Role of intracellular processing events

R Chignola, C Anselmi, A Franceschi, M Pasti, C Candiani, G Tridente and M Colombatti
Institute of Immunology and Infectious Diseases, University of Verona, Italy.

We have assayed the sensitivity of Jurkat cells in different growth phases to an anti-CD5-ricin A chain (ST.1-RTA) immunotoxins (IT). Jurkat cells proliferated exponentially until a stationary growth phase was reached. Proliferating and stationary cells displayed marked differences in sensitivity to ST.1-RTA treatment; the time required to kill one log of target cells (T10) was 70 h in proliferating and 12 h in stationary cells, respectively. Differences in sensitivity to IT treatment were greatly diminished by the addition of the IT enhancer monensin (T10 = 4.9 and 3.5 h in proliferating and stationary cells, respectively). Binding and internalization studies carried out with fluoresceinated ST.1 mAb revealed that the higher sensitivity of stationary cells to ST.1-RTA treatment was not due to an increased uptake or to faster internalization kinetics of IT molecules in this cell population; rather, our data indicated that a different intracellular routing of IT molecules took place in the two cell populations. Mathematical modeling of experimental data allowed us to calculate the efficiency of the intracellular transport of IT molecules toward a subcellular compartment facilitating toxin translocation to the cell cytosol. The IT intracellular processing in stationary cells was 5.5-fold more efficient than in proliferating cells. This value strictly correlated with the higher sensitivity of the stationary cell population to ST.1-RTA treatment.


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R. Chignola, C. Anselmi, M. D. Serra, A. Franceschi, G. Fracasso, M. Pasti, E. Chiesa, J. M. Lord, G. Tridente, and M. Colombatti
Self-potentiation of Ligand-Toxin Conjugates Containing Ricin A Chain Fused with Viral Structures
J. Biol. Chem., October 6, 1995; 270(40): 23345 - 23351.
[Abstract] [Full Text] [PDF]


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