A role for calcium influx in setting the threshold for CD4+CD8+ thymocyte negative selection

We have applied an in vitro system that mimics thymic negative selection to investigate signaling pathways that may be important for the removal of autoreactive cells from the thymus. We sought to more precisely determine the contribution of calcium-dependent pathways to CD4+CD8+ thymocyte deletion that is mediated by either an antigenic peptide or a peptide analogue. We show that the requirement for external calcium influx is dependent upon the strength of the deleting ligand. Furthermore, these results correlate well with a requirement, under certain circumstances, for signaling through the calcium/calmodulin-dependent phosphatase calcineurin. The use of suboptimal stimuli may, therefore, be useful in revealing biochemical pathways important for CD4+CD8+ thymocyte negative selection.