The response of human lymphocytes to phytohemagglutinin is impaired at different levels during aging

Abstract

Several parameters generally believed to be necessary for the activation and progression of proliferation of human lymphocytes have been investigated and compared with special reference to aging. The responding capacity of plasma membrane potential to depolarizing and also repolarizing conditions induced by exposure to mitogens like PHA was lower in lymphocytes from old donors as compared to those of young ones. This indicates a significant age-dependent difference in the readiness to respond to channel-activating perturbations. As an early signal of activation, after one hour PHA stimulation the merocyanine 540 uptake by the lipid regions was chosen, based on the property of this fluorescent probe to bind to loosely packed lipids of the plasma membrane. The proteins encoded by the c-myc and c-myb genes were chosen as markers of the G0/G1 and G1/S phased transition, respectively. The mean number of cells that increased the uptake of MC 540 following mitogenic stimulation did not differ in young vs. old individuals. However, 4 samples out of 10 from the old population showed lower MC 540 fluorescence than the lowest signal from the young population. The number of responding cells was decreased during aging when the presence of the c-myc protein was taken as its measure; and this decrease was further accentuated, determining the expression of the c-myb protein. This frequently encountered age-dependent pattern, however, was not followed by the lymphocytes of all old donors. One example is reported in which the MC 540 uptake, the c-myc and c-myb expression in the cells from one old subject fell in the range of the young subjects. However, even in this case, the response of the lymphocytes as measured by 3H-thymidine incorporation was only 64% of that of young subjects. For this sample, we found an impairment of the response at the mitochondrial level. In addition to these parameters, the amount of 3H-thymidine incorporated by the cells expressing the c-myb protein was calculated. The values in old individuals were lower than those in the young, suggesting that not all the cells expressing the c-myb protein were able to synthesize DNA in lymphocyte populations from the elderly. Our data support the view that the age-dependent decline of lymphocyte responsiveness to mitogens can be accounted for by impairments at different levels.(ABSTRACT TRUNCATED AT 400 WORDS)