Cell Cycle Phase-Specific Survival of CD95 Ligand-Challenged Jurkat Cells: Upregulation of Heat-Shock Response

An important means of regulating T-cell function occurs via physical deletion (cytolysis) of unnecessary/unwanted T cells. Among cytolytic pathways, CD95 (Fas)-based killing plays a prominent role. Although activation of T cells results in rapid upregulation of surface CD95 expression, sensitivity to CD95-based killing lags behind. To assess determinants of resistance to CD95-based killing, we used Jurkat cells as a model. Analysis of the 10% survivors of a LD90 dose of CD95 ligand (CD95L) at 24 h demonstrated them to arise preferentially from the S + G2/M phases of the cell cycle and to remain clustered in S + G2/M without undergoing cell division. Protein immunoblot, immunocytochemistry, and RT-PCR analyses demonstrated that hsp72 was markedly upregulated in CD95L survivors within hours of CD95L challenge, indicative of a heat-shock response. Indeed, exposure of Jurkat cells to bona fide heat shock did markedly upregulate hsp72 and, upon subsequent CD95L challenge, did greatly enhance cell survival with persistent clustering to S + G2/M. These findings collectively suggest that in response to a CD95L insult, development of a heat-shock response above some critical threshold level can protect against lethality. This raises the possibility that exaggerated and/or protracted heat-shock responses under in vivo conditions may favor the survival of T cells (including autoaggressive T cells) that otherwise would be destined to die via a CD95-based pathway.