Mitochondrial ultracondensation, but not swelling, is involved in TNF alpha-induced apoptosis in human T-lymphoblastic leukaemic cells.

Mitochondrial permeability transition (PT) pore opening and mitochondrial swelling have been reported in association with apoptosis. Conformational alterations of mitochondria induced by tumour necrosis factor-alpha (TNF alpha), and the association with TNF alpha-induced apoptosis, were, therefore, studied in the human acute T-lymphoblastic leukaemia (T-ALL) cell line, CCRF-CEM and its vinblastine-resistant CEM/VLB100 cell line by transmission electron microscopy (TEM). The CEM/VLB100 cell line possessed more condensed (C phase) mitochondria in the resting state compared with its parental cell line, consistent with increased activity of the mitochondrial electron transport chain (ETC). Following exposure to TNF alpha, conformational alterations of mitochondria occurred in both apoptotic and non-apoptotic cells. Orthodox (O phase) mitochondria in non-apoptotic cells underwent C-phase, transitional O-phase and slightly swollen (S-phase) conformational changes. TNF alpha-induced mitochondrial swelling was a late event and was found to a far lesser extent than mitochondrial condensation. No swollen mitochondria were observed in apoptotic cells. Ultracondensed (UC phase) mitochondria were observed in cells undergoing both TNF alpha-induced and spontaneous apoptosis and were seen when TNF alpha-induced apoptosis was inhibited by 3-methyladenine (3MA). The structural integrity of UC phase mitochondria persisted through the apoptotic process. We conclude that TNF alpha-induced mitochondrial swelling and apoptosis are separate events. Mitochondrial ultracondensation is associated with the processes signalling apoptosis and is not a result of TNF alpha-induced apoptotic shrinkage.