Spontaneous and stimulated production of cytokines by peripheral blood cells obtained from the caudal vein of male Wistar rats was assessed before testing their resistance to oxygen deficiency in a decompression chamber. To study the spontaneous production of cytokines, heparinized blood cells were incubated in a culture medium (24 h, 5% CO2, 37°C) and the content of proinflammatory cytokines IL-6 and TNFα and anti-inflammatory IL-10 in the culture medium was assessed by ELISA. To stimulate cytokine production, blood cells were incubated for 24 h with LPS, phytohemagglutinin, and concanavalin A at final concentrations of 2, 4, and 4 μg, respectively. Two weeks after blood sampling, individual resistance of the animals to hypoxia in a decompression chamber was determined. In animals with low resistance to hypoxia, the levels of spontaneous production of all three cytokines were significantly higher, while after stimulation, the level of IL-1β increased by more than 2 times. The animals with spontaneous production of IL-10>50 pg/ml, IL-6>10 pg/ml, and TNFα>10 pg/ml, as well as with the increase in IL-1β production by more than 2 times upon stimulation were classified as low-resistant. At IL-10<15 pg/ml, IL-6<9 pg/ml, and TNFα<7 pg/ml, as well as the absence of the increase in IL-1β production upon stimulation, they were classified as high-resistant. The identified features of spontaneous and stimulated production of cytokines can be used as non-invasive biomarkers to determine the resistance to hypoxia without exposure to sublethal hypoxia in a decompression chamber.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Kirova YI, Germanova EL, Lukyanova LD. Phenotypic features of the dynamics of HIF-1α levels in rat neocortex in different hypoxia regimens. Bull. Exp. Biol. Med. 2013;154(6):718-722. https://doi.org/10.1007/s10517-013-2038-z
Dzhalilova DSh, Kosyreva AM, Diatroptov ME, Ponomarenko EA, Tsvetkov IS, Zolotova NA, Mkhitarov VA, Khochanskiy DN, Makarova OV. Dependence of the severity of the systemic inflammatory response on resistance to hypoxia in male Wistar rats. J. Inflamm Res. 2019;12:73-86. https://doi.org/10.2147/JIR.S194581
Jain K, Suryakumar G, Prasad R, Ganju L. Differential activation of myocardial ER stress response: a possible role in hypoxic tolerance. Int. J. Cardiol. 2013;168(5):4667-4677. https://doi.org/10.1016/j.ijcard.2013.07.180
Lukyanova LD, Kirova YuI. Effect of hypoxic preconditioning on free radical processes in tissues of rats with different resistance to hypoxia. Bull. Exp. Biol. Med. 2011;151(3):292-296. https://doi.org/10.1007/s10517-011-1312-1
Julian CG, Subudhi AW, Wilson MJ, Dimmen AC, Pecha T, Roach RC. Acute mountain sickness, inflammation, and permeability: new insights from a blood biomarker study. J. Appl. Physiol. (1985). 2011;111(2):392-399. https://doi.org/10.1152/japplphysiol.00391.2011
Lundeberg J, Feiner JR, Schober A, Sall JW, Eilers H, Bickler PE. Increased cytokines at high altitude: lack of effect of ibuprofen on acute mountain sickness, physiological variables, or cytokine levels. High Alt. Med. Biol. 2018;19(3):249-258. https://doi.org/10.1089/ham.2017.0144
Druzhinina YuG, Ryzhikova SL, Ryabicheva TG, Varaksin NA. Determination of the spontaneous and mitogen-induced cytokine production levels in conditionally healthy donors ex vivo. Med. Immunol. 2009;11(4-5):473. Russian.
Golderova AS, Nikolaeva IN, Tarasova VE, Kozlov VA. Features of cytokine production in patients with unstable angina: dependence on ethnicity in the Republic of Sakha (Yakutia). Med. Immunol. 2012;14(1-2):143-148. Russian. https://doi.org/10.15789/1563-0625-2012-1-2-143-148
Kosyreva AM, Dzhalilova DSh, Tsvetkov IS, Makarova MA, Makarova OV. Ex vivo production of IL-1β and IL-10 by activated blood cells of Wistar rats with different resistance to hypoxia after systemic inflammatory response syndrome. Bull. Exp. Biol. Med. 2023;176(2):290-296. https://doi.org/10.1007/s10517-024-06010-5
Movafagh A, Heydary H, Mortazavi-Tabatabaei SA, Azargashb E. The significance application of indigenous phytohemagglutinin (PHA) mitogen on metaphase and cell culture procedure. Iran. J. Pharm. Res. 2011;10(4):895-903.
Kang SS, Kim SK, Baik JE, Ko EB, Ahn KB, Yun CH, Han SH. Staphylococcal LTA antagonizes the B cell-mitogenic potential of LPS. Sci. Rep. 2018;8(1):1496. https://doi.org/10.1038/s41598-018-19653-y
Nonis SG, Haywood J, Schmidberger JW, Mackie ERR, Soares da Costa TP, Bond CS, Mylne JS. Structural and biochemical analyses of concanavalin A circular permutation by jack bean asparaginyl endopeptidase. Plant Cell. 2021;33(8):2794-2811. https://doi.org/10.1093/plcell/koab130
Lee SH, Lee H, Park R. Systemic immune modulation induced by ephedrine in obese-diabetes (db/db) mice. Curr. Issues Mol. Biol. 2023;45(12):10097-10108. https://doi.org/10.3390/cimb45120630
Dzhalilova D, Kosyreva A, Lokhonina A, Tsvetkov I, Vishnyakova P, Makarova O, Fatkhudinov T. Molecular and phenotypic distinctions of macrophages in tolerant and susceptible to hypoxia rats. PeerJ. 2023;11:e16052. https://doi.org/10.7717/peerj.16052
Savilova AM, Chulkina MM, Alexeev LP. Comparative investigation of interleukin-2 and interleukin-2 receptor alpha mRNA expression in lymphocytes activated by PHA or CONA. Immunologiya. 2013;(2):76-80. Russian.
Taylor CT, Scholz CC. The effect of HIF on metabolism and immunity. Nat. Rev. Nephrol. 2022;18(9):573-587. https://doi.org/10.1038/s41581-022-00587-8
Author information
Authors and Affiliations
Corresponding author
Additional information
Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 177, No. 4, pp. 419-423, April, 2024
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Dzhalilova, D.S., Kosyreva, A.M. & Makarova, O.V. Spontaneous and Stimulated Production of Cytokines by Blood Cells Ex Vivo as a Biomarker of Initially High or Low Hypoxia Resistance in Rats. Bull Exp Biol Med (2024). https://doi.org/10.1007/s10517-024-06200-1
Received:
Published:
DOI: https://doi.org/10.1007/s10517-024-06200-1