Cytotoxic T-cell-derived granzyme B activates the apoptotic protease ICE-LAP3

Cytotoxic T lymphocytes (CTLs) and natural killer (NK) cells provide immune surveillance against viruses and neoplasms, and play a central role in the pathogenesis of autoimmune disease, AIDS and graft rejection [1,2]. Thus, it is important to understand the precise molecular mechanism(s) whereby cytotoxic lymphocytes destroy susceptible target cells, Granule-mediated cytotoxicity requires a combination of both perforin and granzyme B [3]. Perforin polymerizes to form transmembrane channels and presumably allows granzyme B access to target cell substrates, which until recently, were unknown, One clue to the identity of the physiological substrate(s) activated by granzyme B comes from its unusual specificity for cleaving synthetic substrates after aspartate residues [4], Members of the ICE/CED-3 family of cysteine proteases are prime candidates as they are important apoptotic effecters [5,6] and are expressed as zymogens, which can be processed to form active heterodimeric enzymes after cleavage at specific aspartate residues, Previous studies have shown that granzyme B proteolytically activates the cell death effector Yama/CPP32/apopain [7,8] (referred to here as Yama), Here we report that granzyme B also activates ICE-LAP3/Mch3/CMH-1 (referred to here as ICE-LAP3), which, along with Yama and Mch2, forms a subset of the ICE/CED-3 family of cysteine proteases most closely related to the Caenorhabditis elegans cell death gene, CED-3 [6,9], Importantly, Jurkat T cells incubated with granzyme B and a sublytic concentration of perforin undergo apoptosis, which is preceded by the activation of endogenous ICE-LAP3, Thus, we propose that granzyme B mediates apoptosis by directly engaging the target cell's death effector machinery, which is probably composed of an arsenal of intracellular, CED-3-like cysteine proteases.