Aerobic glycolysis is dispensable for pro-inflammatory cytokine production by dendritic cells

• Anna S. Budikhina
• Nina E. Murugina
• Polina V. Maksimchik
• Dagil Yu.A.
• Melnikov M.V.
• Balyasova L.S.
• Vladimir V. Murugin
• Chkadua G.Z.
• Boris V. Pinegin
• Mikhail V. Pashenkov

Abstract

Introduction. Activation of innate immune cells is accompanied by an increase in glycolysis; however, the links between glycolysis and specific parameters of innate immune response have not been conclusively established.

Aims – to study the relationship between glycolysis and proinflammatory cytokine production by dendritic cells (DCs) activated by lipopolysaccharide.

Material and methods. DCs were generated from blood monocytes of healthy donors and stimulated by LPS in the presence of different combinations of D-glucose, 2-deoxy-D-glucose (2-DG) and D-mannose. Cytokine production was analysed by enzyme-linked immunosorbent assay, cytokine gene expression by real-time PCR with reverse transcription, glycolysis by XFe96 real-time metabolism analyzer.

Results. We show that aerobic glycolysis is dispensable for pro-inflammatory cytokine production by human DCs in vitro. Upon activation with LPS, macrophages and DCs produce comparable amounts of the pro-inflammatory cytokines tumor necrosis factor (TNF) and interleukin-6 (IL-6), despite the much lower rate of glycolysis in DCs. Inhibition of glycolysis by glucose starvation (culturing in glucose-free medium) does not significantly affect TNF, IL-6 and IL-12p70 production by LPS-activated DCs. At the same time, 2-deoxy-D-glucose (2-DG), a widely used glycolysis inhibitor, suppresses production of these cytokines by LPS-activated DCs and compromises their viability. Unlike glucose starvation, 2-DG induces an unfolded protein response (UPR). Cancellation of this UPR by an excess of D-mannose alleviates negative effects of 2-DG on DC cytokine production and viability.

Conclusion. Obtained data allow to partially revise current views on the role of glycolysis in the development of innate immune response.

Keywords:dendritic cells; macrophages; glycolysis; 2-deoxy-D-glucose; lipopolysaccharide; cytokines; unfolded protein response

References

1. Kadowaki N., Ho S., Antonenko S., Malefyt R.W., Kastelein R.A., Bazan F., et al. Subsets of human dendritic cell precursors express different toll-like receptors and respond to different microbial antigens. J. Exp. Med. 2001; 194 (6): 863–9.

2. Khaitov R.M., Pinegin B.V., Pashenkov M.V. Role of pattern recognition receptors in innate and adaptive immunity. Immunologiya. 2009; 30 (1): 66–76. (in Russian)

3. Everts B., Amiel E., Huang S.C., Smith A.M., Chang C.H., Lam W.Y., et al. TLR-driven early glycolytic reprogramming via the kinases TBK1-IKKepsilon supports the anabolic demands of dendritic cell activation. Nat. Immunol. 2014; 15 (4): 323–32.

4. Everts B., Amiel E., van der Windt G.J., Freitas T.C., Chott R., Yarasheski K.E., et al. Commitment to glycolysis sustains survival of NO-producing inflammatory dendritic cells. Blood. 2012; 120 (7): 1422–31.

5. Rodriguez-Prados J.C., Traves P.G., Cuenca J., Rico D., Aragones J., Martin-Sanz P., et al. Substrate fate in activated macrophages: a comparison between innate, classic, and alternative activation. J. Immunol. 2010; 185 (1): 605–14.

6. Na Y.R., Gu G.J., Jung D., Kim Y.W., Na J., Woo J.S., et al. GM-CSF induces inflammatory macrophages by regulating glycolysis and lipid metabolism. J. Immunol. 2016; 197 (10): 4101–9.

7. Murugina N.E., Balyasova L.S., Budikhina A.S., Maximchik P.V., Dagil Yu.A., Murugin V.V., Chkadua G.Z., Pinegin B.V., Pashenkov M.V. Metabolic reprogramming of macrophages upon activation with a NOD1 receptor agonist. Immunologiya. 2019; 40 (1): 5–14. (in Russian)

8. Dietl K., Renner K., Dettmer K., Timischl B., Eberhart K., Dorn C., et al. Lactic acid and acidification inhibit TNF secretion and glycolysis of human monocytes. J. Immunol. 2010; 184 (3): 1200–9.

9. Hu K., Yang Y., Lin L., Ai Q., Dai J., Fan K., et al. Caloric restriction mimetic 2-deoxyglucose alleviated inflammatory lung injury via suppressing nuclear pyruvate kinase M2-signal transducer and activator of transcription 3 pathway. Front. Immunol. 2018; 9: 426.

10. Tannahill G.M., Curtis A.M., Adamik J., Palsson-McDermott E.M., McGettrick A.F., Goel G., et al. Succinate is an inflammatory signal that induces IL-1beta through HIF-1alpha. Nature. 2013; 496 (7444): 238–42.

11. O’Neill L.A., Pearce E.J. Immunometabolism governs dendritic cell and macrophage function. J. Exp. Med. 2015; 213 (1): 15–23.

12. Xi H., Kurtoglu M., Lampidis T.J. The wonders of 2-deoxy-D-glucose. IUBMB Life. 2014; 66 (2): 110–21.

13. Maximchik P., Abdrakhmanov A., Inozemtseva E., Tyurin-Kuzmin P.A., Zhivotovsky B., Gogvadze V. 2-Deoxy-D-glucose has distinct and cell line-specific effects on the survival of different cancer cells upon antitumor drug treatment. FEBS J. 2018; 285 (24): 4590–601.

14. Grootjans J., Kaser A., Kaufman R.J., Blumberg R.S. The unfolded protein response in immunity and inflammation. Nat. Rev. Immunol. 2016; 16 (8): 469–84.

15. Smith J.A. Regulation of cytokine production by the unfolded protein response; implications for infection and autoimmunity. Front. Immunol. 2018; 9: 422.

16. Budikhina A.S., Murugina N.E., Maximchik P.V., Dagil Yu.A., Nikolaeva A.M., Balyasova L.S., Murugin V.V., Chkadua G.Z., Pinegin B.V., Pashenkov M.V. On the role of glycolysis in pro-inflammatory cytokine production by macrophages. Immunologiya. 2019; 40 (5): 11–22. (in Russian)

17. Pashenkov M.V., Balyasova L.S., Dagil Y.A., Pinegin B.V. The role of the p38-MNK-eIF4E signaling axis in TNF production downstream of the NOD1 receptor. J Immunol. 2017; 198 (4): 1638–48.

18. Sallusto F., Lanzavecchia A. Efficient presentation of soluble antigen by cultured human dendritic cells is maintained by granulocyte/macrophage colony-stimulating factor plus interleukin 4 and downregulated by tumor necrosis factor alpha. J. Exp. Med. 1994; 179 (4): 1109–18.

19. O’Neill L.A., Kishton R.J., Rathmell J. A guide to immunometabolism for immunologists. Nat. Rev. Immunol. 2016; 16 (9): 553–65.

20. Jackson R.J., Hellen C.U., Pestova T.V. The mechanism of eukaryotic translation initiation and principles of its regulation. Nat. Rev. Mol. Cell Biol. 2010; 11 (2): 113–27.

21. Langenkamp A., Messi M., Lanzavecchia A., Sallusto F. Kinetics of dendritic cell activation: impact on priming of TH1, TH2 and nonpolarized T cells. Nat. Immunol. 2000; 1 (4): 311–6.

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