Non-transferrin dependent 59Fe uptake in phytohemagglutinin-stimulated human peripheral lymphocytes

Human peripheral lymphocytes stimulated by phytohemagglutinin (PHA) have been used to demonstrate the characteristics of iron uptake from non-transferrin iron donors. When incubated with 59Fe(II)-ascorbate or 59Fe(III)-nitrilotriacetate (Fe-NTA), stimulated lymphocytes (2 micrograms/ml PHA) showed a tenfold increase in uptake of 59Fe as compared with the resting cells. The uptake of 59Fe from these iron donors was time and concentration dependent, showed saturation kinetics, and was not influenced by the addition of a tenfold excess of unlabeled Fe(III)2-transferrin (Fe-Tf). The amount of 59Fe accumulated from 59Fe-NTA was about one-third of that from 59Fe2-Tf, whereas the uptake of 59Fe from 59Fe-ascorbate was about tenfold higher. When stimulated with varying doses of PHA, lymphocytes showed maximal uptake of 59Fe from 59Fe2-Tf at the concentration of PHA (1 microgram/ml) optimal for 3H-thymidine incorporation. Lymphocytes stimulated with supraoptimal concentrations (5-20 micrograms/ml) of PHA showed slightly less iron uptake from 59Fe-Tf and had a longer transferrin cycle time, as compared with the cells under optimal stimulation. In contrast, the uptake of 59Fe from non-transferrin 59Fe donors in cells stimulated with 5-20 micrograms/ml PHA was greater than that in cells grown with 1 or 2 micrograms/ml PHA. It is suggested that lymphocytes take up iron from non-transferrin iron donors by processes different from the iron uptake pathway used by transferrin.