Institute of Science and the Environment, University of Worcester, Worcestershire, United Kingdom.
Institute of Sport, Physical Activity and Leisure, Leeds Beckett University , Leeds , United Kingdom.
J Appl Physiol (1985). 2018 Aug 1;125(2):586-595. doi: 10.1152/japplphysiol.01137.2017. Epub 2018 Apr 12.
This study investigated immunophenotypic differences in intracellular thiol redox state of peripheral blood mononuclear cells (PBMCs) isolated from trained [ n = 9, means ± SD: age 28 ± 5 yr; (body mass index) BMI 23.2 ± 2.6 kg/m; V̇o (maximal oxygen intake)56.9 ± 6.1 ml·kg·min] and recreationally active (RA, n = 11, means ± SD: age 27 ± 6 yr; BMI 24.2 ± 3.7 kg/m; V̇o 45.1 ± 6.4 ml·kg·min) participants before and after a maximal aerobic exercise tolerance test. Blood samples were taken before (Pre), during (sample acquired at 70% maximum heart rate), immediately after (Post + 0), and 15 min postexercise (Post + 15). PBMCs were isolated, and reduced thiol analysis [fluorescein-5 maleimide (F5M)] by immunophenotype [cluster of differentiation (CD)3, CD4, and CD8] was performed using flow cytometry. A significant increase in cellular F5M fluorescence was observed in CD3 T cells at Post + 0, with changes driven to a greater extent by CD8 T cells (fold change in both groups CD4: +2.3, CD8: +2.8; P < 0.05). Further analysis revealed a population of highly reduced CD8 T cells (CD8T-reduced) that significantly increased from Pre to Post + 0 in RA participants only (RA: +272 cell/µl, P < 0.05). To understand these results further, CD8T-reduced and CD8T-reduced cells were analyzed for immunophenotype in response to the same exercise protocol ( n = 6, means ± SD: age 24 ± 5 yr; BMI 25.7 ± 4.1 kg·m; V̇o 41.33 ± 7.63 ml·kg·min). CD8T-reduced had significantly less lymphoid homing potential (chemokine receptor type 7) Post + 0 compared with Pre. This study is the first, to our knowledge, to demonstrate that lymphocyte populations become more reductive in response to acute exercise. NEW & NOTEWORTHY The study presented provides the first evidence to suggest that cytotoxic T cells become transiently reductive in healthy individuals following a single bout of cycling. Detection of these cells was enabled via the use of a flow cytometric assay that incorporates the thiol reactive probe fluorescein-5 maleimide. Using this method, transient reductive stress in viable T cells is permissible and provides the basis for further research in the area of exercise immunology.
这项研究调查了训练有素的[n=9，平均值±标准差：年龄 28±5 岁；（体重指数）BMI 23.2±2.6 kg/m；V̇o（最大摄氧量）56.9±6.1 ml·kg·min]和有规律的业余活动（RA，n=11，平均值±标准差：年龄 27±6 岁；BMI 24.2±3.7 kg/m；V̇o 45.1±6.4 ml·kg·min）参与者在最大有氧耐力测试前后外周血单个核细胞（PBMCs）细胞内硫醇氧化还原状态的免疫表型差异。在测试前（Pre）、进行中（在最大心率的 70%时获得样本）、运动后即刻（Post + 0）和运动后 15 分钟（Post + 15）时采集血液样本。使用流式细胞术通过免疫表型[分化群（CD）3、CD4 和 CD8]进行 PBMCs 的还原硫分析[荧光素 5 马来酰亚胺（F5M）]。在 Post + 0 时，CD3 T 细胞中的细胞 F5M 荧光显著增加，变化主要由 CD8 T 细胞驱动（两组的变化倍数 CD4：+2.3，CD8：+2.8；P<0.05）。进一步的分析显示，RA 参与者中只有 CD8 T 细胞（CD8T-减少）的一个高度还原的群体从 Pre 到 Post + 0 显著增加（RA：+272 个细胞/μl，P<0.05）。为了进一步了解这些结果，对 6 名参与者（平均值±标准差：年龄 24±5 岁；BMI 25.7±4.1 kg·m；V̇o 41.33±7.63 ml·kg·min）进行了 CD8T-减少和 CD8T-减少细胞对相同运动方案的免疫表型分析。与 Pre 相比，Post + 0 时 CD8T-减少的淋巴细胞归巢潜力（趋化因子受体 7 型）显著降低。本研究首次证明，淋巴细胞群体在急性运动后变得更具还原性。新的和值得注意的是，该研究提供了第一个证据，表明健康个体在单次循环运动后，细胞毒性 T 细胞会暂时变得还原性。通过使用结合硫醇反应探针荧光素 5 马来酰亚胺的流式细胞术检测到这些细胞。使用这种方法，在存活的 T 细胞中允许短暂的还原应激，并为运动免疫学领域的进一步研究提供了基础。
