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3‐Deaza‐Adenosine Inhibition of Stimulus‐Response Coupling in Human Polymorphonuclear Leukocytes

In an effort to define better the functional role of S‐adenosyl‐methionine‐mediated methylation reactions in modulating polymorphonuclear (PMN) functional responses to chemotactic stimuli, we investigated the effects of 3‐deaza‐adenosine (3‐DZA), a known inhibitor of methylation reactions in phagocytic cells, on formyl methionyl‐leucyl‐phenylalanine (FMLP)‐induced responses in human PMN leukocytes. Using the fluorescent cyanine dye 3,3'‐dipropylthiocarbocyanine (di‐S‐C3‐(5)) as an optical probe of membrane potential we observed that 3‐DZA at concentrations that inhibit FMLP‐induced O2− production does not significantly alter FMLP‐induced changes in transmembrane potential. Additional studies showed an inhibitory effect of 3‐DZA on FMLP‐induced PMN pinocytosis and to a lesser degree on FMLP‐induced degranulation. However, pretreatment of PMNs with 3‐DZA did not alter FMLP‐induced changes in Quin‐2 fluorescence, an indicator of changes in intracellular calcium levels. These findings demonstrate a dissociation between chemotactic factor‐induced cell membrane depolarization, changes in intracellular calcium, and specific neutrophil functional responses and suggest that chemotactic factor‐induced changes in transmembrane potential and intracellular calcium are independent of chemotactic factor‐induced methylation reactions. Furthermore, 3‐DZA did not alter phorbol myristate acetate induced O2− production or fluid pinocytosis indicating a stimulus specificity for the inhibitory effects of this agent on O2− production.