Reversible cell damage by T-cell perforins. Calcium influx and propidium iodide uptake into K562 cells in the absence of lysis.
The non-lethal effects of the lymphocyte-derived pore-forming toxin perforin on the human erythroleukaemia cell line K562 were investigated. By using the fluorescent Ca2+ indicator fura-2, perforin was shown to cause intracellular Ca2+ concentration to rise transiently into the micromolar range in the absence of cell death. By fluorescence-activated cell sorting it was demonstrated that K562 cells took up the membrane-impermeant nuclear stain propidium iodide (PI) when exposed to non-lethal doses of perforin. The permeability to PI was short-lived, confirming the transience of the perforin pore. Analogies with non-lethal effects and recovery processes occurring in nucleated cells exposed to the membrane-attack complex of complement are drawn.
Jones
J
J
Department of Medical Biochemistry, University of Wales College of Medicine, Heath Park, Cardiff, U.K.
Hallett
M B
MB
Morgan
B P
BP
D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D002118:Calcium / Q000378:metabolism*; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D006801:Humans; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive; D008562:Membrane Glycoproteins; D008565:Membrane Proteins / Q000302:isolation & purification / Q000494:pharmacology*; D051379:Mice; D054353:Perforin; D010617:Phenanthridines / Q000378:metabolism*; D052899:Pore Forming Cytotoxic Proteins; D011419:Propidium / Q000378:metabolism*; D013602:T-Lymphocytes, Cytotoxic; D014407:Tumor Cells, Cultured / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*
D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D010617:Phenanthridines; D052899:Pore Forming Cytotoxic Proteins; D054353:Perforin; D011419:Propidium; D002118:Calcium