To investigate the effect of intense exercise on immune function, 12 healthy males (26 +/- 1 yr) underwent cycle exercise to exhaustion at 80% maximum work load. One hour later, six of the subjects underwent a second identical bout. Blood was drawn preexercise (C), at exhaustion (Ex-1, Ex-2), and at 1 h of recovery (Rec-1, Rec-2). At Ex-1 and Ex-2, total leukocytes (monocytes, neutrophils, and lymphocytes) increased significantly (P less than 0.05), and all returned to C levels by Rec-1 except lymphocytes, which were lower than at C. At Rec-2 total leukocytes remained higher (P less than 0.05) than at C, primarily because of elevated neutrophil counts. Phenotype analysis indicated a 2.5-fold increase in CD16+ (natural killer) cells at Ex-1 and a decrease primarily in CD4+ (T-helper) cells. All phenotype changes returned to C levels at Rec-1. At Ex-1 the in vitro mitogenic response to concanavalin A, phorbol myristate acetate + ionomycin, phytohemagglutinin, and pokeweed mitogen decreased (P less than 0.05) but returned to levels not different from C at Rec-1. Both the in vitro percent specific lysis of a target natural killer cell (K562) and the lytic activity per cell increased (P less than 0.05) in peripheral mononuclear cells at Ex-1, returning to C levels at Rec-1. The total leukocyte counts with exercise were significantly correlated with plasma epinephrine concentration, and a causal effect is thus possible. Such acute changes in numbers and function of circulating cells of the immune system may possibly have functional and clinical correlates.