Enhancement of mitogen response and surface marker analysis of lymphocytes from young and old donors after preliminary incubation in vitro

We have examined the detailed kinetics of PHA induced proliferation of freshly isolated mononuclear cells from young and old human donors. Our studies confirm that donors over the age of 60 years may have a decreased number of PHA responsive cells and that these cells have a significantly diminished rate of entry into the first cell cycle. Age of the donor does not affect the time at which significant numbers of cells appear in first S-phase, the duration of first S-phase, the doubling time in exponential growth, and thymidine uptake per cell. Cells from young and old donors were also cultured in medium supplemented with pooled human serum for 1 or 2 days prior to the PHA response assay. After 2 days of preliminary culture, the PHA response in both young and old is significantly enhanced, with a greater enhancement in the old. The basis of this enhancement appears to be a significant increase in rate of entry into the first cell cycle. None of the other kinetic parameters were significantly altered. The magnitude of the rate enhancement in old donors' cells eliminated the difference in entry rate between young and old after 2 days of preliminary culture. The same rate enhancing effects were seen in the presence of serum pooled from young or old donors. There is no statistically significant change in the number of responding cells after preliminary culture. We had previously suggested, based on lactate dehydrogenase (LD) subunit ratio and patterns of T cell associated surface markers, that less differentiated subpopulations of T cells exist in elderly donors. After preliminary culture, no significant change was seen in the proportions of cells positive for T3, T4, T8, T10 and Ia surface antigens and the unusual pattern of surface marker distribution was still present on the old donors' cells. It appears that the greater rate enhancement in old donors' cells after preliminary culture may not be due to induced maturation of the possibly less differentiated T cell populations described previously. The results do suggest that these subpopulations may be non-responsive to PHA.