Garcia-Souza 2017 MiP2017

Short-term adaptation to high altitude does not affect respiration of peripheral blood mononuclear cells and platelets.

Link: MiP2017

Garcia-Souza Luiz F, Velika B, Sumbalova Z, Menz V, Gatterer H, Volani C, Lundby C, Burtscher M, Gnaiger E (2017)

Event: MiP2017

Peripheral-blood mononuclear cells (PBMC) and platelets (PLT) are potentially powerful models for the diagnosis of mitochondrial respiratory function and dysfunction, offering a minimally invasive approach in comparison to tissue biopsies. However, several variables that may interfere with mitochondrial physiology in blood cells are still unsolved. High altitude experience induces whole body adaptations due to low O₂ concentration in the atmosphere and cold temperature. Decrease of plasma volume, increase in arterial blood pressure and red blood cells count are some of the adaptations regarding the blood stream adaptation to altitude [1,2]. However, mitochondrial function of PBMC and PLT under altitude adaptation was never addressed. Therefore, the objective of our study is to analyse mitochondrial function of PBMC and PLT exposed to 2020 m altitude adaptation during the first 48 hours.

Blood was collected from eight healthy female volunteers (26.3 ± 5.7 years) at low altitude of 575 m (Innsbruck, Austria) for 4 days and exposed to altitude of 2020 m (Kühtai, Austria) for 2 days. During the study, the participants were under a controlled diet and physical activity restricted to a light 1-h walk per day. Every morning at fasted state, blood was taken (18 mL) from up to 4 participants for measurements of freshly isolated blood cell respiration. PBMC and PLT were separated from whole blood by centrifugation on Ficoll-Paque™ PLUS density medium using 50 mL Leucosep tubes [3]. After the first washing of PBMC-platelets layer with DPBS (25 mL, 120 g, RT), the PBMC were washed two times with ice-cold DPBS+2% FBS (50 mL, 300 g, 4 °C). Isolated cells were counted on SYSMEX XN-350 haematology analyser. 3·106 PBMC or 150·106 PLT were added into the 2 mL chambers of the O2k-FluoRespirometer (Oroboros Instruments, Innsbruck, Austria) containing mitochondrial respiration medium MiR05-Kit with catalase at 37 °C [3,4]. The coupling control & viability protocol (CCVP) was applied. Respiration of PBMC was corrected for the contribution by contaminating platelets.

PBMC showed a ROUTINE respiration of 5.0 amol·s^-1·cell^-1 at 575 m and 5.2 amol·s^-1·cell^-1 at 2020 m. ET capacity was 18.8 amol·s^-1·cell^-1 (575 m) and 17.4 amol·s^-1·cell^-1 (2020 m). PLT showed a ROUTINE respiration of 0.12 amol·s^-1·cell^-1 at 575 m and 0.11 amol·s^-1·cell^-1 at 2020 m. ET capacity was 0.34 amol·s^-1·cell^-1 (575 m) and 0.31 amol·s^-1·cell^-1 (2020 m). There were no significant differences in altitude adaptation in PBMC and PLT. Flux control ratios in both cell types were not affected by altitude adaptation and cell viability was not compromised from altitude effect or any apparent complication during sample transport, 94% and 93.3% in PBMC (575 m and 2020 m, respectively) and 92.2% and 90.5% in PLT (575 m and 2020 m, respectively).

In summary, PBMC and PLT respiration was not affected during the first 48 hours of altitude experience. According to our results, altitude adaptation will not be a variable which interferes in mitochondrial function diagnostics in pathological states during the first 48 hours of altitude exposure. However, longer exposure of altitude adaptation or higher altitude exposure should be investigated.

• Bioblast editor: Kandolf G • O2k-Network Lab: AT Innsbruck Oroboros, SK Bratislava Sumbalova Z, AT Innsbruck Burtscher M, CH Zurich Lundby C, CH Zurich University of Zurich Physiology, DK Copenhagen Lundby C, AT Innsbruck Gnaiger E

Labels: MiParea: Respiration 

Organism: Human  Tissue;cell: Blood cells, Platelet 

Coupling state: ROUTINE, ET-pathway"ET-pathway" is not in the list (LEAK, ROUTINE, OXPHOS, ET) of allowed values for the "Coupling states" property. 

HRR: Oxygraph-2k 

PMBCs 

Affiliations and support

Garcia-Souza Luiz F(1,2), Velika B(2,3), Sumbalova Z(2,4), Menz V(1), Gatterer H(1), Volani C(5), Lundby C(6), Burtscher M(1), Gnaiger E(2,7)

1. Inst Sport Science, Univ Innsbruck, Austria
2. Daniel Swarovski Research Lab, Dept Visceral, Transplant Thoracic Surgery, Medical Univ Innsbruck, Austria
3. Dept Medical Clinical Biochem, Fac Medicine, Pavol Jozef Šafárik Univ Košice, Slovakia
4. Pharmacobiochemical Lab, 3rd Dept Internal Medicine, Fac Medicine, Comenius Univ, Bratislava, Slovakia
5. Dept Internal Medicine II, Medical Univ Innsbruck, Austria
6. Center Physical Activity Research, Univ Hospital Copenhagen, Denmark
7. Oroboros Instruments, Innsbruck, Austria. – [email protected]

Supported by K-Regio project MitoFit (GSLF, ZS, VM) and Action Austria-Slovakia (BV). Contribution to European Union Framework Programme Horizon 2020 COST Action CA15203 MITOEAGLE.

References

1. Siebenmann C, Robach P, Lundby C (2017) Regulation of blood volume in lowlanders exposed to high altitude. J Appl Physiol, jap-00118.
2. Mazzeo RS (2008) Physiological responses to exercise at altitude. Sports Med 38:1-8.
3. Garcia-Souza LF, Velika B, Sumbalova Z, Menz V, Burtscher M, Gnaiger E (2017) Assessment of mitochondrial respiratory function in cryopreserved platelets. Abstract, MiPschool Obergurgl 2017.
4. Sumbalova Z, Hiller E, Chang S, Garcia L, Droescher S, Calabria E, Volani C, Krumschnabel G, Gnaiger E (2016) Isolation of blood cells for HRR. Mitochondr Physiol Network 21.17:1-15.