Addition of the chemotactic peptide, f-Met-Leu-Phe, to human monocytes induced a burst of superoxide release, which ceased after approximately 3 min. Diminished responsiveness to f-Met-Leu-Phe, but not to phorbol myristate acetate (PMA), was induced by 1- to 3-h storage at 0 degrees C or by 2 min in 40 microM adenosine (ADO). Reversal of the ADO block was achieved by addition of adenosine deaminase (ADA) as little as 15 sec before the f-Met-Leu-Phe stimulus; ADA had no effect when added poststimulus. The ADO experiments suggest that there are a minimum of two sequentially produced intermediates in the f-Met-Leu-Phe stimulus-response pathway. The first intermediate persists for less than 30 sec. The second, formation of which is stimulated by the first, persists for the duration of the response and is the target of ADO inhibition. The ADO target is apparently not protein kinase-C, since the response of inhibited cells to PMA was unimpaired. The maximal inhibition by adenosine of f-Met-Leu-Phe-induced superoxide generation was approximately 50%. It is possible that f-Met-Leu-Phe stimulates two pathways of NADPH activation, only one of which is inhibited by adenosine.