Phytohemagglutinin induced changes of membrane lipid packing, c-myc and c-myb encoded protein expression in human...

Three parameters which signal different stages of cell activation were analyzed in lymphocytes from young and old subjects. Merocyanine 540 (MC-540) incorporation into the membrane lipid phase was used as a very early marker of activation and was measured after 1 h of phytohemagglutinin (PHA) stimulation. The proteins coded by c-myc and c-myb protooncogenes were determined by appropriate antibodies and were taken as markers of the G0/G1 and G1/S phase transition, respectively. The number of cells which increased the uptake of MC-540 following PHA stimulation did not differ when comparing young and old individuals. Both the number of the responding cells and the size of the response were decreased during aging when the presence of the c-myc protein was taken into account. A consistent decrease of the percentage of lymphocytes able to express the c-myb protein was observed in the cells from old donors as compared to those from the young ones, but the amount of detectable protein per cell remained unchanged. Our data suggest that the deficiency of responsiveness which accompanies aging is due to impairments at different points of the cell cycle. The very low number of cells expressing the c-myb protein is likely the result of step by step elimination of those cells not able to fulfill the requirements to progress along the cell cycle.