Longevity of human allospecific TLCs: mycoplasma infection as a cause of in vitro "suppression" of MLC.

It has been suggested that allospecific T-cell clones lose specific reactivity after approximately 30 cell doublings and subsequently acquire suppressor and NK-like characteristics. We have tested this hypothesis by assaying paired functional and nonfunctional TLCs for suppressor activity in PLT and MLC cocultures. Two sets of clones were initially studied: the first pair consisted of clone S5.2B, a functional TLC, and S5.14A, a nonfunctional TLC; the second pair of clones tested was comprised of two different expansions of the same clone S5.5A (nonfunctional) and S5.5B (functional). These experiments yielded no evidence for suppressive activity by nonfunctional clones toward functional clones, furthermore, the addition of nonfunctional clones to primary MLC assays had no effect on the level of responsiveness. Eight clones were subcloned and 89 subclones were retested for function after approximately 50 cell doublings. Generally, the subclones failed to suppress MLC proliferation. A minority of TLCs could suppress MLC responses, but this "suppression" was reversible with the addition of 2% exogenous TCGF. However, eight subclones and two parental TLC lines did suppress MLC responses in the presence or absence of TCGF, but the suppressive effects in such cocultures were reversible in the presence of tylocine, an anti-mycoplasma antibiotic. Therefore, human T-cells, cultured for extended periods, do not inexorably and universally lose specific alloreactivity and gain suppressive characteristics due to some presumed differentiative event.