During extreme exercise, athletes experience increased inflammation that is similar to the acute phase response. Endurance athletes, distance runners in particular, are also more susceptible to compromised iron stores. This study evaluated inflammation, immune function and iron status in athletes completing a 50K ultramarathon. Twenty-two well-trained distance runners, 11 males and 11 females, were randomized in a double blind manner into--1) those who consumed 300 mg vitamin E and 1000 mg vitamin C (500 mg twice daily) or 2) placebos--for six weeks before and one week following a 50K ultramarathon race. Blood samples were obtained on 13 separate occasions throughout the study: before supplementation, during supplementation, the day before the race, pre-race, mid-race, immediately post-race, 2 hours following the race, and daily for six days following the race. Plasma levels of ascorbic acid and ��-tocopherol were measured by HPLC with electrochemical detection. Inflammatory cytokines, interleukin-6 (IL-6), tumor necrosis factor-�� (TNF-��), and interleukin-1�� (IL-1��) were measured using standard clinical assays. Each subject recorded immune function in an activity log and incidence of illness was tabulated as number of days ill. Ferritin was measured by enzyme immunoassay. Hemoglobin, hematocrit, and total-iron binding capacity (TIBC) and serum total iron were analyzed by standard procedures. Plasma concentrations of ascorbic acid and ��-tocopherol increased significantly in supplemented subjects (p<0.0001). Although the ultramarathon race elicited an inflammatory response, antioxidant supplementation did not alter the responses of IL-6 and TNF-��, which both increased from pre-race to mid-race, post- and post-2 h (Scheffe post-hoc analysis, p<0.0001) and returned to pre-race concentrations by 1 day after the race. Male supplemented subjects had lower IL-1�� concentrations compared to females consuming the supplement or to males consuming the placebo (ANCOVA, gender/time/treatment interaction; p<0.01) at mid-race (p<0.05 females, p<0.005 males), post 1 and 2 days (all p<0.002). Males had significantly higher ferritin levels than the female subjects (ANOVA, p<0.0001); supplementation resulted in lower ferritin concentrations at post-5 days (p<0.02, ANCOVA treatment time interaction, p<0.005). Supplementation did not reduce the days illness among those consuming antioxidants compared to those consuming the placebos. Ferritin not only increases during inflammation, it also is a measure of iron stores. Females had significantly lower levels of iron than the male subjects for each of the iron parameters measured (hemoglobin and hematocrit both p<0.0001, ferritin p<0.001, TIBC p<0.02) excluding serum total iron. The ferritin concentrations measured in the women were indicative of depleted iron stores (<12 ��g/l), and antioxidant supplementation increased hematocrit levels in the female subjects (p<0.05). This investigation indicates that female distance runners need to be aware of an increased susceptibility to iron depletion compared to their male counterparts. Antioxidant supplementation improved hematocrit levels (p<0.05) among female runners and may improve iron status among females with depleted stores. Although other investigations have suggested that antioxidant vitamins decrease exercise induced inflammation, no profound benefit of supplementation was found in this investigation though a response similar to the acute phase response was elicited by the ultramarathon race. Improvements in IL-i and ferritin in response to antioxidant supplementation may indicate that the supplementation was beneficial, but more research is needed to draw definitive conclusions. / Graduation date: 2003