Long-Term in vitro Growth of Human T Cell Clones: Can Postmitotic ‘Senescent’ Cell Populations Be Defined?

The long-term in vitro growth characteristics of peripheral, nonspecifically expanded T cell clones from young (< 30 years old) and old (> 65 years old) Senieur-protocol compatible healthy blood donors were compared. Additionally, T cell clones derived from the sites of autoimmune events were studied. Until the 6th week in culture all clones showed a progressive increase in cell number. CD25 expression and proliferation in response to stimulation with recombinant IL-2 remained stable. Staining of DNA fragments with propidium iodide, however, revealed a small percentage of apoptotic cells. After 7-12 weeks in culture a rapid decrease in cell numbers accompanied by a striking increase in the number of apoptotic cells was noted in all peripheral clones. Although CD25 was still present on almost 100% of the cells, receptor density and proliferative response to IL-2 were greatly reduced. After a maximum of 14 weeks in culture, corresponding to 29 population doublings (PD), all peripheral T cell clones had died. No difference was noted between CD4 and CD8 T cell clones, but clones from old donors tended to have a shorter lifespan and a lower number of PD than clones from young persons (n.s.). Interestingly, T cells from autoimmune lesions had a longer lifespan and a higher number of PD than peripheral Tcell clones (p < 0.01). In contrast to peripheral T cell clones, which died almost immediately after reaching the end of their proliferative capacity, a small but stable nondividing cell population could be distinguished in autoimmune clones. Although these cells had lost their capacity to divide in response to IL-2 stimulation, they still expressed the low affinity chain of the IL-2 receptor, at the same intensity as at the initiation of the cultures. These results (a) confirm previous data that peripheral T cell clones have a limited lifespan; (b) suggest that this may be due to an imbalance between mitosis and apoptosis, and (c) enable the postulation that T lymphocytes from autoimmune lesions may, at least partly, escape this regulatory mechanism.