Transferrin binding to peripheral blood lymphocytes activated by phytohemagglutinin involves a specific receptor. Ligand interaction.
Immunohistological studies have indicated that membrane sites binding transferrin are present upon activated human peripheral blood lymphocytes. In this study, we have investigated transferrin uptake in human lymphocytes exposed to phytohemagglutinin (PHA), by quantitative radiobinding and immunofluorescence in parallel. In stimulated lymphocytes, binding was maximal after a 30-min incubation, being greatest at 37 degrees C, and greater at 22 degrees C than at 4 degrees C. Although some shedding and endocytosis of transferrin occurred at 22 degrees and 37 degrees C, these factors, and resulting synthesis of new sites, did not affect measurement of binding which was found to be saturable, reversible, and specific for transferrin (K(a) 0.5-2.5 x 10(8) M(-1)). Binding was greater after a 48-h exposure to PHA than after 24 h, and was maximal at 66 h. Sequential Scatchard analysis revealed no significant elevation in affinity of interaction. However, although the total number of receptors increased, the proportion of cells in which binding of ligand was detected immunohistologically increased in parallel, and after appropriate correction, the cellular density of receptors remained relatively constant throughout (60,000-80,000 sites/cell). Increments in binding during the culture period were thus due predominantly to expansion of a population of cells bearing receptors. Similar differences in binding were apparent upon comparison of cells cultured in different doses of PHA, and in unstimulated cells binding was negligible. Transferrin receptors appear, therefore, to be readily detectable only upon lymphocytes that have been activated.
D004305:Dose-Response Relationship, Drug; D005455:Fluorescent Antibody Technique; D006801:Humans; D007501:Iron / Q000378:metabolism; D008214:Lymphocytes / Q000378:metabolism*; D010835:Phytohemagglutinins / Q000494:pharmacology*; D011869:Radioligand Assay / Q000379:methods; D011956:Receptors, Cell Surface / Q000378:metabolism*; D011990:Receptors, Transferrin; D014168:Transferrin / Q000378:metabolism*
D010835:Phytohemagglutinins; D011956:Receptors, Cell Surface; D011990:Receptors, Transferrin; D014168:Transferrin; D007501:Iron