Exercise-induced ß2-adrenergic Receptor Activation Enhances the Antileukemic Activity of Expanded γδ T-Cells via DNAM-1 Upregulation and PVR/Nectin-2 Recognition.

Exercise mobilizes cytotoxic lymphocytes to blood which may allow superior cell products to be harvested and manufactured for cancer therapy. Gamma-Delta (γδ) T-cells have shown promise for treating solid tumors, but there is a need to increase their potency against hematologic malignancies. Here, we show that human γδ T-cells mobilized to blood in response to just 20 minutes of graded exercise have surface phenotypes and transcriptomic profiles associated with cytotoxicity, adhesion, migration, and cytokine signaling. Following 14 days ex vivo expansion with zoledronic acid and IL2, exercise mobilized γδ T-cells had surface phenotypes and transcriptomic profiles associated with enhanced effector functions and demonstrated superior cytotoxic activity against multiple hematologic tumors in vitro and in vivo in leukemia-bearing xenogeneic mice. Infusing humans with the ß1+ß2-agonist isoproterenol and administering ß1 or ß1+ß2 antagonists prior to exercise revealed these effects to be ß2-adrenergic receptor (AR) dependent. Antibody blocking of DNAM-1 on expanded γδ T-cells, as well as the DNAM-1 ligands PVR and Nectin-2 on leukemic targets, abolished the enhanced antileukemic effects of exercise. These findings provide a mechanistic link between exercise, ß2-AR activation, and the manufacture of superior γδ T-cell products for adoptive cell therapy against hematologic malignancies. SIGNIFICANCE: Exercise mobilizes effector γδ T-cells to blood via ß2-adrenergic signaling which allows for generation of a potent expanded γδ T-cell product that is highly cytotoxic against hematologic malignancies.

Acute exercise increases immune responses to SARS CoV-2 in a previously infected man.

Evidence is emerging that exercise and physical activity provides protection against severe COVID-19 disease in patients infected with SARS-CoV-2, but it is not known how exercise affects immune responses to the virus. A healthy man completed a graded cycling ergometer test prior to and after SARS-CoV-2 infection, then again after receiving an adenovirus vector-based COVID-19 vaccine. Using whole blood SARS-CoV-2 peptide stimulation assays, IFN-γ ELISPOT assays, flow cytometry, ex vivo viral-specific T-cell expansion assays and deep T-cell receptor (TCR) ß sequencing, we found that exercise robustly mobilized highly functional SARS-CoV-2 specific T-cells to the blood compartment that recognized spike protein, membrane protein, nucleocapsid antigen and the B.1.1.7 α-variant, and consisted mostly of CD3+/CD8+ T-cells and double-negative (CD4-/CD8-) CD3+ T-cells. The magnitude of SARS-CoV-2 T-cell mobilization with exercise was intensity dependent and robust when compared to T-cells recognizing other viruses (e.g. CMV, EBV, influenza). Vaccination enhanced the number of exercise-mobilized SARS-CoV-2 T-cells recognizing spike protein and the α-variant only. Exercise-mobilized SARS-CoV-2 specific T-cells proliferated more vigorously to ex vivo peptide stimulation and maintained broad TCR-ß diversity against SARS-CoV-2 antigens both before and after ex vivo expansion. Neutralizing antibodies to SARS-CoV-2 were transiently elevated during exercise after both infection and vaccination. Finally, infection was associated with an increased metabolic demand to defined exercise workloads, which was restored to pre-infection levels after vaccination. This case study provides impetus for larger studies to determine if these immune responses to exercise can facilitate viral clearance, ameliorate symptoms of long COVID syndrome, and/or restore functional exercise capacity following SARS-CoV-2 infection.

Variations in strength-related measures during the menstrual cycle in eumenorrheic women: A systematic review and meta-analysis.

OBJECTIVES: To systematically review the current body of research that has investigated changes in strength-related variables during different phases of the menstrual cycle in eumenorrheic women. DESIGN: Systematic review and meta-analysis. METHODS: A literature search was conducted in Pubmed, SPORTDiscus and Web of Science using search terms related to the menstrual cycle and strength-related measures. Two reviewers reached consensus that 21 studies met the criteria for inclusion. Methodological rigour was assessed using the Quality Assessment Tool for Observational Cohort and Cross-Sectional Studies. Random effects meta-analyses were used to compare the early-follicular, ovulatory and mid-luteal phases for maximal voluntary contraction, isokinetic peak torque, and explosive strength. RESULTS: The assessment of study quality showed that a high level of bias exists in specific areas of study design. Non-significant and small or trivial effect sizes (p≥0.26, Hedges g≤0.35) were identified for all strength-related variables in each comparison between phases. 95% confidence intervals for each comparison suggested the uncertainty associated with each estimate extends to a small effect on strength performance with unclear direction (-0.42≤g≤0.48). The heterogeneity for each comparison was also small (p≥0.83, I2=0%). CONCLUSIONS: Strength-related measures appear to be minimally altered (g≤0.35) by the fluctuations in ovarian sex hormones that occur during the menstrual cycle. This finding should be interpreted with caution due to the methodological shortcomings identified by the quality assessment.

Are There Benefits from the Use of Fish Oil Supplements in Athletes? A Systematic Review.

Despite almost 25 y of fish oil supplementation (FS) research in athletes and widespread use by the athletic community, no systematic reviews of FS in athletes have been conducted. The objectives of this systematic review are to: 1) provide a summary of the effect of FS on the athlete's physiology, health, and performance; 2) report on the quality of the evidence; 3) document any side effects as reported in the athlete research; 4) discuss any risks associated with FS use; and 5) provide guidance for FS use and highlight gaps for future research. Electronic databases (PubMed, Embase, Web of Science, Google Scholar) were searched up until April 2019. Only randomized placebo-controlled trials (RCTs) in athletes, assessing the effect of FS on a health, physiological/biochemical, or performance variable were included. Of the 137 papers identified through searches, 32 met inclusion criteria for final analysis. Athletes varied in classification from recreational to elite, and from Olympic to professional sports. Mean age for participants was 24.9 ± 4.5 y, with 70% of RCTs in males. We report consistent effects for FS on reaction time, mood, cardiovascular dynamics in cyclists, skeletal muscle recovery, the proinflammatory cytokine TNF-α, and postexercise NO responses. No clear effects on endurance performance, lung function, muscle force, or training adaptation were evident. Methodological quality, applying the Physiotherapy Evidence Database (PEDro) scale, ranged from 6 to a maximum of 11, with only 4 RCTs reporting effect sizes. Few negative outcomes were reported. We report various effects for FS on the athlete's physiology; the most consistent findings were on the central nervous system, cardiovascular system, proinflammatory cytokines, and skeletal muscle. We provide recommendations for future research and discuss the potential risks with FS use.

Systemic ß-Adrenergic Receptor Activation Augments the ex vivo Expansion and Anti-Tumor Activity of Vγ9Vδ2 T-Cells.

TCR-gamma delta (γδ) T-cells are considered important players in the graft-vs.-tumor effect following allogeneic hematopoietic cell transplantation (alloHCT) and have emerged as candidates for adoptive transfer immunotherapy in the treatment of both solid and hematological tumors. Systemic ß-adrenergic receptor (ß-AR) activation has been shown to mobilize TCR-γδ T-cells to the blood, potentially serving as an adjuvant for alloHCT and TCR-γδ T-cell therapy. We investigated if systemic ß-AR activation, using acute dynamic exercise as an experimental model, can increase the mobilization, ex vivo expansion, and anti-tumor activity of TCR-γδ T-cells isolated from the blood of healthy humans. We also sought to investigate the ß-AR subtypes involved, by administering a preferential ß1-AR antagonist (bisoprolol) and a non-preferential ß1 + ß2-AR antagonist (nadolol) prior to exercise as part of a randomized placebo controlled cross-over experiment. We found that exercise mobilized TCR-γδ cells to blood and augmented their ex vivo expansion by ~182% compared to resting blood when stimulated with IL-2 and ZOL for 14-days. Exercise also increased the proportion of CD56+, NKG2D+/CD62L-, CD158a/b/e+ and NKG2A- cells among the expanded TCR-γδ cells, and increased their cytotoxic activity against several tumor target cells (K562, U266, 221.AEH) in vitro by 40-60%. Blocking NKG2D on TCR-γδ cells in vitro eliminated the augmented cytotoxic effects of exercise against U266 target cells. Furthermore, administering a ß1 + ß2-AR (nadolol), but not a ß1-AR (bisoprolol) antagonist prior to exercise abrogated the exercise-induced enhancement in TCR-γδ T-cell mobilization and ex vivo expansion. Furthermore, nadolol completely abrogated while bisoprolol partially inhibited the exercise-induced increase in the cytotoxic activity of the expanded TCR-γδ T-cells. We conclude that acute systemic ß-AR activation in healthy donors markedly augments the mobilization, ex vivo expansion, and anti-tumor activity of TCR-γδ T-cells and that some of these effects are due to ß2-AR signaling and phenotypic shifts that promote a dominant activating signal via NKG2D. These findings highlight ß-ARs as potential targets to favorably alter the composition of allogeneic peripheral blood stem cell grafts and improve the potency of TCR-γδ T-cell immune cell therapeutics.

Iron balance and iron supplementation for the female athlete: A practical approach.

Maintaining a positive iron balance is essential for female athletes to avoid the effects of iron deficiency and anaemia and to maintain or improve performance. A major function of iron is in the production of the oxygen and carbon dioxide carrying molecule, haemoglobin, via erythropoiesis. Iron balance is under the control of a number of factors including the peptide hormone hepcidin, dietary iron intake and absorption, environmental stressors (e.g. altitude), exercise, menstrual blood loss and genetics. Menstruating females, particularly those with heavy menstrual bleeding are at an elevated risk of iron deficiency. Haemoglobin concentration [Hb] and serum ferritin (sFer) are traditionally used to identify iron deficiency, however, in isolation these may have limited value in athletes due to: (1) the effects of fluctuations in plasma volume in response to training or the environment on [Hb], (2) the influence of inflammation on sFer and (3) the absence of sport, gender and individually specific normative data. A more detailed and longitudinal examination of haematology, menstrual cycle pattern, biochemistry, exercise physiology, environmental factors and training load can offer a superior characterisation of iron status and help to direct appropriate interventions that will avoid iron deficiency or iron overload. Supplementation is often required in iron deficiency; however, nutritional strategies to increase iron intake, rest and descent from altitude can also be effective and will help to prevent future iron deficient episodes. In severe cases or where there is a time-critical need, such as major championships, iron injections may be appropriate.

Salt-sensitive hypertension: mechanisms and effects of dietary and other lifestyle factors.

Salt sensitivity, which is an increase in blood pressure in response to high dietary salt intake, is an independent risk factor for cardiovascular disease and mortality. It is associated with physiological, environmental, demographic, and genetic factors. This review focuses on the physiological mechanisms of salt sensitivity in populations at particular risk, along with the associated dietary factors. The interplay of mechanisms such as the renin-angiotensin aldosterone system, endothelial dysfunction, ion transport, and estrogen decrease in women contributes to development of salt sensitivity. Because of their effects on these mechanisms, higher dietary intakes of potassium, calcium, vitamin D, antioxidant vitamins, and proteins rich in L-arginine, as well as adherence to dietary patterns similar to the DASH (Dietary Approaches to Stop Hypertension) diet, can be beneficial to salt-sensitive populations. In contrast, diets similar to the typical Western diet, which is rich in saturated fats, sucrose, and fructose, together with excessive alcohol consumption, may exacerbate salt-sensitive changes in blood pressure. Identifying potential mechanisms of salt sensitivity in susceptible populations and linking them to protective or harmful dietary and lifestyle factors can lead to more specific guidelines for the prevention of hypertension and cardiovascular disease.

Critical Difference and Biological Variation in Biomarkers of Oxidative Stress and Nutritional Status in Athletes.

UNLABELLED: The longitudinal monitoring of oxidative stress (OS) in athletes may enable the identification of fatigued states and underperformance. The application of OS biomarker monitoring programs in sport are hindered by reliability and repeatability of in-the-field testing tools, the turnaround of results, and the understanding of biological variation (BV). Knowledge of BV and critical difference values (CDV) may assist with data interpretation in the individual athlete. METHODS: We aimed firstly to assess the repeatability of the clinical point of care redox test, Free Oxygen Radical Test (FORT) and the Free Oxygen Radical Defence (FORD) in trained participants and elite athletes and secondly to calculate the analytical, BV, CDV and index of individuality (II) for FORT, FORD, red blood cell glutathione, lutein, α and γ-tocopherol. Part 1: Fifteen elite athletes were sampled in duplicate for calculation of the repeatability of the FORT and FORD tests. Part 2: Twelve well-trained athletes had venous samples drawn every 2 hours from 0800 to 1800 for calculation of BV, CDV, II for FORT, FORD, RBC GSH, lutein, α-tocopherol and γ-tocopherol. RESULTS: Repeatability of the FORT and FORD assay was 3.9% and 3.7% respectively. Biomarker CDV ranged from 12.8% to 37%, with a circadian effect for FORT, α-tocopherol and γ-tocopherol (p<0.01), with all biomarker indices of individuality < 0.8 arbitrary units. CONCLUSION: We report that the use of the novel redox test in athletes is practical, and the generation of BV and CDV for biomarkers of OS enhances the interpretation of physiologically meaningful changes in individuals above the use of clinical reference ranges alone.

Is iron treatment beneficial in, iron-deficient but non-anaemic (IDNA) endurance athletes? A systematic review and meta-analysis.

PURPOSE: The aim of this study was to determine whether iron treatments improve the iron status and aerobic capacity of iron deficient non-anaemic endurance athletes. METHOD: A meta-analysis of studies that investigated the effects of iron treatment on serum ferritin (sFer), serum iron (sFe), transferrin saturation (Tsat), haemoglobin concentration ([Hb]) and (VO(2max)). Seventeen eligible studies were identified from online databases. RESULTS: Analysis of pooled data indicated that iron treatments had a large effect on improving sFer (Hedges' g=1.088, 95% CI 0.914 to 1.263, p<0.001), sFe (Hedges' g=1.004, 95% CI 0.828 to 1.181, p<0.001) and Tsat (Hedges g=0.741, 95% CI 0.564 to 0.919, p<0.001) and a moderate effect on improving [Hb] (Hedges' g=0.695, 95% CI 0.533 to 0.836, p<0.001) and (VO(2max)) (Hedges' g=0.610, 95% CI 0.399 to 0.821, p<0.001). Regression analysis revealed a significant interaction between the effect of iron treatment on sFer and treatment duration, suggesting treatments that lasted beyond 80 days appear to have the least effect on sFer. CONCLUSIONS: These results indicate iron treatments improve the iron status and aerobic capacity of iron deficient non-anaemic endurance athletes.

Effect of Intravenous Iron on Aerobic Capacity and Iron Metabolism in Elite Athletes.

PURPOSE: Iron-deficient athletes are often treated with long-term, low-dose iron therapy. Such treatments may be efficacious in correcting iron deficiency; however, the effect on acute and chronic iron metabolism and subsequent endurance capacity is less clear. METHODS: Fifteen national and international standard runners were identified as iron deficient nonanemic (IDNA) and assigned to either an intravenous iron treatment group or placebo group. Participants completed three exercise tests to volitional exhaustion, as follows: before treatment, within 24 h, and 4 wk after treatment. RESULTS: Serum ferritin, serum iron, and transferrin saturation were significantly improved in the iron group after intervention and compared with those in placebo (P < 0.05). Hepcidin levels were significantly greater before and after exercise after the iron injection (P < 0.05), and this was independent of changes in interleukin-6. There were no differences between groups in red cell indices, total hemoglobin mass, V˙O2max, submaximal blood lactate, running economy, RPE, or time to exhaustion (P > 0.05). CONCLUSIONS: A single 500-mg intravenous iron injection is effective for improving iron status for at least 4 wk, but this does not lead to improved aerobic capacity. This investigation suggests that iron availability supersedes inflammation in the regulation of hepcidin in IDNA endurance athletes after acute intravascular iron injection treatment.

A case study of an iron-deficient female Olympic 1500-m runner.

This case study examines the impact of low serum ferritin (sFe) on physiological assessment measures and performance in a young female 1500-m runner undertaking approximately 95-130 km/wk training. The study spans 4 race seasons and an Olympic Games. During this period, 25 venous blood samples were analyzed for sFe and hemoglobin (Hb); running economy, VO(2max), and lactate threshold were measured on 6 occasions separated by 8-10 mo. Training was carefully monitored including 65 monitored treadmill training runs (targeting an intensity associated with the onset of blood lactate accumulation) using blood lactate and heart rate. Performances at competitive track events were recorded. All data were compared longitudinally. Mean sFe was 24.5 ± 7.6 µg/L (range 10-47), appearing to be in gradual decline with the exception of 2 data points (37 and 47 µg/L) after parenteral iron injections before championships, when the lowest values tended to occur, coinciding with peak training volumes. Each season, 1500-m performance improved, from 4:12.8 in year 1 to 4:03.5 in year 4. VO(2max) (69.8 ± 2.0 mL · kg(-1) · min(-1)) and running economy (%VO(2max) at a fixed speed of 16 km/h; max 87.8%, min 80.3%) were stable across time and lactate threshold improved (from 14 to 15.5 km/h). Evidence of anemia (Hb <12 g/dL) was absent. These unique data demonstrate that in 1 endurance athlete, performance can continue to improve despite an apparent iron deficiency. Raising training volume may have caused increased iron utilization; however, the effect of this on performance is unknown. Iron injections were effective in raising sFe in the short term but did not appear to affect the long-term pattern.