ACTIVATION EVENTS DURING THYMIC SELECTION

During their differentiation in the mouse thymus, CD4+8- cells undergo several of the sequential changes observed upon normal activation of mature, peripheral CD4+ lymphocytes. Expression of CD69, an early activation marker, is first observed on a minority of cells at the T cell receptor (TCR)lo/med double-positive stage, is maximal (50-90%) on heat-stable antigen (HSA)hi TCR(hi) double-positive, HSA(hi) TCR(med) CD4+8lo, and HSA(hi) TCR(hi) CD4+8- cells, and is downmodulated at the mature HSA(lo) CD4+8- stage. In contrast, CD44, a late activation marker, is selectively expressed at the HSA(lo) stage. The set of lymphokines that CD4+8-thymocytes can produce upon stimulation also characteristically expands from mainly interleukin 2 (IL-2) at the HSA(hi) stage, to IL-2 and very large amounts of IL-4, IL-5, IL-10, and interferon-gamma IFN-gamma) at the HSA(lo) stage. 1 in 30 HSA(lo) CD4+8- adult thymocytes secrete IL-4 upon stimulation through their TCR. This frequency is 25% of the frequency of IL-2 producers, about 100-fold above that of peripheral (mainly resting) CD4+ T cells. With time after their generation in organ culture, CD4+8- thymocytes lose their capacity to secrete IL-4, IL-5, and IFN-gamma, but not IL-2. Similarly, the frequency of IL-4, but not of IL-2, producers progressively decreases after emigration to the periphery as judged by direct comparison between thymic and splenic CD4+ cells in newborns, or by following the fate of intrathymically labeled CD4+8-cells in adults after their migration to the spleen. This sequence suggests that thymic selection results from an activation process rather than a simple rescue from death at the double-positive stage, and shows that the functional changes induced after intrathymic activation, although transient, are still evident after export to the periphery.