Supplementary MaterialsAdditional file 1: Protocols for T cell and Monocyte phenotypes. emphasis on understanding how different forms of exercise affect immune function. Mechanistic studies are beginning to shed light on how exercise may impair the development of cancer or be used to augment cancer treatment. The beneficial effects of exercise on the immune system may be exploited to improve patient responses to cancer immunotherapy. Methods We investigated the effects of acute exercise on the composition of peripheral blood leukocytes over time in a male population of varying fitness. Subjects performed a brief maximal intensity cycling regimen and a longer less intense cycling regimen at separate visits. Leukocytes were measured by multi-parameter flow cytometry of more than 50 immunophenotypes for each collection sample. Results We found a differential induction of leukocytosis dependent on exercise intensity and duration. Cytotoxic natural killer cells demonstrated the greatest increase Roscovitine inhibition (average of 5.6 fold) immediately post-maximal exercise whereas CD15+ granulocytes demonstrated the largest increase at 3 h post-maximal exercise (1.6 fold). The longer, less intense endurance exercise resulted in an attenuated leukocytosis. Induction of leukocytosis did not differ in our limited study of active ( 0.01 by the Bonferronis multiple comparison post-test of all pairs Having identified the cell types that changed upon exercise, we tested the effects of two types of exercise on the magnitude of induction of leukocytosis for distinct leukocyte subsets. Figure?2a shows the change in six of the eight previously shown immune cell populations (lymphocytes and total CD3+ cells not shown) from baseline to post exercise and from baseline to 3 h post exercise. CD56+CD16+ natural killer cells (NK cells) showed the greatest degree of change versus other cell populations in both the maximal and endurance regimens Roscovitine inhibition (Bartletts test for equal variances em p /em ? ?0.0001; em p /em ? ?0.01 for NK cell compared to other cell populations). NK cell induction was significantly higher in the maximal test versus the endurance test (5.6 fold increase vs 2.8 respectively; em p /em ? ?0.0001). CD56brCD16? NK cells, thought to be precursor cells to CD56+CD16+ NK cells Rabbit Polyclonal to YOD1 , did not increase as much as CD56+CD16+ NK cells (2.7 fold in maximal and 1.8 in endurance; p?=?0.0017 and p?=?0.029 respectively) Roscovitine inhibition (Fig.?2b). We looked at the degree of change in other cell types. Another cell type involved in immune surveillance, TCR+ T cells, was induced after maximal exercise (2.6 fold) (Fig.?2b). Although monocytes were induced to a small degree upon exercise, we investigated whether there was differential induction in three monocyte subsets (Classical- CD14+CD16?; Intermediate- CD14+CD16+; Non-classical- CD14loCD16+) . Immediately post maximal exercise; nonclassical monocytes were induced to the highest degree and were higher than classical monocytes (2.7 fold compared to 1.6; em p /em ? ?0.01) (Fig.?2c). As observed with other cell Roscovitine inhibition types, there was no difference in the attenuated induction after the endurance exercise. These data highlight the distinct coordination of the mobilization of immune cells in response to different modes of exercise. Open in a separate window Fig. 2 Natural killer cells show the greatest degree of induction upon exercise. a The values for each subset at the Post or 3HR time point were divided by the Pre sample value. Box and whisker plots show the 25th and 75th percentiles (box) with median (line in box) and the minimum and maximum values. b The changes from Pre to Post for CD56++CD16? NK cells and T cells. c Changes in the distribution of Classical (CD14+CD16?), Intermediate (CD14+CD16+), and Non-classical monocytes (CD14loCD16+). The means comparing Pre versus Post were significantly different ( em p /em ? ?0.0001 by repeated measures ANOVA). **?=? em p /em ? ?0.01 by the Bonferronis multiple comparison post-test of all pairs The peripheral blood leukocyte system responds distinctly to different types of exercise To illustrate the peripheral blood leukocyte system changes in response to exercise, we took the mean values of each major phenotype from the 15 subjects and created pie graphs to reflect the size and composition of the leukocytes at each time point (Fig.?3a). The baseline sample prior to the maximal test was used to set the pie graph at 100% and the subsequent pie graphs were all sized in proportion to the baseline sample. As a whole, maximal exercise resulted in the expansion of cells at immediately post exercise (163%) and remained high at the three hour time point (135%) whereas leukocytes demonstrated slower kinetics with a 122% increase immediately after exercise and peaking to Roscovitine inhibition 140% at the three hour time point for the endurance test. After 24 h, the system returned to baseline for both regimens (98% for maximal and 100% for endurance). From another viewpoint, the.