Phorbol 12-myristate 13-acetate induced cell death of porcine peripheral blood polymorphonuclear leucocytes

Abstract

Polymorphonuclear leucocytes (PMNs) circulating in mammalian peripheral blood are terminally differentiated cells, and once isolated in serum-free medium, they undergo apoptosis within 1 or 2 days. In this study, we studied the effects of phorbol myristate acetate (PMA) on the viability of porcine PMNs in vitro. PMA is known to suppress apoptosis in many cell types. PMA but not dioctanoyl glycerol (DOG) induced morphological degeneration and cell death within 3 to 5 hours as assessed by light microscopy observation and the MTT viability assay. This occurred despite the fact that DNA fragmentation associated with 'spontaneous apoptosis' was not observed. Morphological degeneration and death were not due to the oxidative damage from superoxides or its metabolites produced by polymorphonuclear leucocytes, because PMA and DOG similarly stimulated superoxide production. Several other inactive phorbol derivatives tested did not cause cell death, suggesting that the toxicity of PMA did not result from non-specific effect of the reagent.