Inflammation Research

, Volume 66, Issue 4, pp 353–364 | Cite as

Adenosine effectively restores endotoxin-induced inhibition of human neutrophil chemotaxis via A1 receptor-p38 pathway

  • Xiaohan Xu
  • Shuyun Zheng
  • Yuyun Xiong
  • Xu Wang
  • Weiting Qin
  • Huafeng Zhang
  • Bingwei SunEmail author
Original Research Paper


Neutrophil chemotaxis plays an essential role in recruiting neutrophils to sites of inflammation. Neutrophil chemotaxis is suppressed both after exposure to lipopolysaccharide (LPS) in vitro and during clinical and experimental endotoxemia, leading to serious consequences. Adenosine (ADO) is a potent anti-inflammatory agent that acts on a variety of neutrophil functions. However, its effects on human neutrophil chemotaxis during infection have been less well characterized. In the present study, we investigated the effect of ADO and its receptor-specific antagonist and agonist on neutrophil chemotaxis in an in vitro LPS-stimulated model. The results showed that increasing the concentration of ADO effectively restored the LPS-inhibited neutrophil chemotaxis to IL-8. A similar phenomenon occurred after intervention with a selective A1 receptor agonist but not with a selective antagonist. Pre-treatment with cAMP antagonist failed to restore LPS-inhibited chemotaxis. Furthermore, protein array and western blot analysis showed that the activation of A1 receptor significantly decreased LPS-induced p38 MAPK phosphorylation. However, the surface expression of the A1 receptor in LPS-stimulated neutrophils was not significantly changed. Taken together, these data indicated that ADO restored the LPS-inhibited chemotaxis via the A1 receptor, which downregulated the phosphorylation level of p38 MAPK, making this a promising new therapeutic strategy for infectious diseases.


Neutrophil Adenosine Chemotaxis Lipopolysaccharide P38 



This study was supported by the National Natural Science Foundation of China, No. 81071546, No. 81272148, No. 81171786, No. 81471903 and No. 81301657.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no competing interests.


  1. 1.
    Wagner JG, Roth RA. Neutrophil migration during endotoxemia. J Leukoc Biol. 1999;66(1):10–24.PubMedGoogle Scholar
  2. 2.
    Barletta KE, Ley K, Mehrad B. Regulation of neutrophil function by adenosine. Arterioscler Thromb Vasc Biol. 2012;32(4):856–64.CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Luster AD. Chemokines–chemotactic cytokines that mediate inflammation. N Engl J Med. 1998;338(7):436–45.CrossRefPubMedGoogle Scholar
  4. 4.
    Itoh Y, Okanoue T. Chemotactic cytokines (chemokines) in human hepatitis and experimental hepatitis models: which ones play the crucial role? J Gastroenterol. 2000;35(9):724–5.CrossRefPubMedGoogle Scholar
  5. 5.
    Heit B, et al. An intracellular signaling hierarchy determines direction of migration in opposing chemotactic gradients. J Cell Biol. 2002;159(1):91–102.CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Heit B, et al. PTEN functions to ‘prioritize’ chemotactic cues and prevent ‘distraction’ in migrating neutrophils. Nat Immunol. 2008;9(7):743–52.CrossRefPubMedGoogle Scholar
  7. 7.
    Pham CT. Neutrophil serine proteases: specific regulators of inflammation. Nat Rev Immunol. 2006;6(7):541–50.CrossRefPubMedGoogle Scholar
  8. 8.
    Maderazo EG, et al. Polymorphonuclear leukocyte migration abnormalities and their significance in seriously traumatized patients. Ann Surg. 1983;198(6):736–42.CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Berger D, et al. Incidence and pathophysiological relevance of postoperative endotoxemia. FEMS Immunol Med Microbiol. 1995;11(4):285–90.CrossRefPubMedGoogle Scholar
  10. 10.
    Matsuura M. Structural modifications of bacterial lipopolysaccharide that facilitate gram-negative bacteria evasion of host innate immunity. Front Immunol. 2013;4:109.CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Putker F, Bos MP, Tommassen J. Transport of lipopolysaccharide to the Gram-negative bacterial cell surface. FEMS Microbiol Rev. 2015;39(6):985–1002.CrossRefPubMedGoogle Scholar
  12. 12.
    Shimazu R, et al. MD-2, a molecule that confers lipopolysaccharide responsiveness on Toll-like receptor 4. J Exp Med. 1999;189(11):1777–82.CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Bohmer RH, Trinkle LS, Staneck JL. Dose effects of LPS on neutrophils in a whole blood flow cytometric assay of phagocytosis and oxidative burst. Cytometry. 1992;13(5):525–31.CrossRefPubMedGoogle Scholar
  14. 14.
    Bishop NC, et al. Pre-exercise carbohydrate status and immune responses to prolonged cycling: I. Effect on neutrophil degranulation. Int J Sport Nutr Exerc Metab. 2001;11(4):490–502.CrossRefPubMedGoogle Scholar
  15. 15.
    Guthrie LA, et al. Priming of neutrophils for enhanced release of oxygen metabolites by bacterial lipopolysaccharide. Evidence for increased activity of the superoxide-producing enzyme. J Exp Med. 1984;160(6):1656–71.CrossRefPubMedGoogle Scholar
  16. 16.
    Akgul C, Moulding DA, Edwards SW. Molecular control of neutrophil apoptosis. FEBS Lett. 2001;487(3):318–22.CrossRefPubMedGoogle Scholar
  17. 17.
    Harkness RA, Simmonds RJ, Coade SB. Purine transport and metabolism in man: the effect of exercise on concentrations of purine bases, nucleosides and nucleotides in plasma, urine, leucocytes and erythrocytes. Clin Sci (Lond). 1983;64(3):333–40.CrossRefGoogle Scholar
  18. 18.
    Bours MJ, et al. Adenosine 5′-triphosphate and adenosine as endogenous signaling molecules in immunity and inflammation. Pharmacol Ther. 2006;112(2):358–404.CrossRefPubMedGoogle Scholar
  19. 19.
    Hasko G, et al. Adenosine receptors: therapeutic aspects for inflammatory and immune diseases. Nat Rev Drug Discov. 2008;7(9):759–70.CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Jacobson KA, Gao ZG. Adenosine receptors as therapeutic targets. Nat Rev Drug Discov. 2006;5(3):247–64.CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Fresco P, et al. Release inhibitory receptors activation favours the A2A-adenosine receptor-mediated facilitation of noradrenaline release in isolated rat tail artery. Br J Pharmacol. 2002;136(2):230–6.CrossRefPubMedPubMedCentralGoogle Scholar
  22. 22.
    Gessi S, et al. The A3 adenosine receptor: an enigmatic player in cell biology. Pharmacol Ther. 2008;117(1):123–40.CrossRefPubMedGoogle Scholar
  23. 23.
    Quinn MT, DeLeo FR, Bokoch GM. Neutrophil methods and protocols. Preface. Methods Mol Biol. 2007;412:vii–viii.PubMedGoogle Scholar
  24. 24.
    Wang X, et al. Exogenous carbon monoxide inhibits neutrophil infiltration in LPS-induced sepsis by interfering with FPR1 via p38 MAPK but not GRK2. Oncotarget. 2016;7(23):34250–65.PubMedPubMedCentralGoogle Scholar
  25. 25.
    Janetopoulos C, Firtel RA. Directional sensing during chemotaxis. FEBS Lett. 2008;582(14):2075–85.CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Devreotes P, Janetopoulos C. Eukaryotic chemotaxis: distinctions between directional sensing and polarization. J Biol Chem. 2003;278(23):20445–8.CrossRefPubMedGoogle Scholar
  27. 27.
    Dilao R, Hauser MJ. Chemotaxis with directional sensing during Dictyostelium aggregation. C R Biol. 2013;336(11–12):565–71.CrossRefPubMedGoogle Scholar
  28. 28.
    Murdoch C, Finn A. Chemokine receptors and their role in inflammation and infectious diseases. Blood. 2000;95(10):3032–43.PubMedGoogle Scholar
  29. 29.
    Perez-Aso M, et al. Adenosine A2A receptor and TNF-alpha regulate the circadian machinery of the human monocytic THP-1 cells. Inflammation. 2013;36(1):152–62.CrossRefPubMedPubMedCentralGoogle Scholar
  30. 30.
    Swain SD, et al. Inhibition of the neutrophil NADPH oxidase by adenosine is associated with increased movement of flavocytochrome b between subcellular fractions. Inflammation. 2003;27(1):45–58.CrossRefPubMedGoogle Scholar
  31. 31.
    Salmon JE, Cronstein BN. Fc gamma receptor-mediated functions in neutrophils are modulated by adenosine receptor occupancy. A1 receptors are stimulatory and A2 receptors are inhibitory. J Immunol. 1990;145(7):2235–40.PubMedGoogle Scholar
  32. 32.
    van der Hoeven D, et al. A role for the low-affinity A2B adenosine receptor in regulating superoxide generation by murine neutrophils. J Pharmacol Exp Ther. 2011;338(3):1004–12.CrossRefPubMedPubMedCentralGoogle Scholar
  33. 33.
    McColl SR, et al. Immunomodulatory impact of the A2A adenosine receptor on the profile of chemokines produced by neutrophils. FASEB J. 2006;20(1):187–9.PubMedGoogle Scholar
  34. 34.
    Inoue Y, et al. A3 and P2Y2 receptors control the recruitment of neutrophils to the lungs in a mouse model of sepsis. Shock. 2008;30(2):173–7.PubMedPubMedCentralGoogle Scholar
  35. 35.
    Jordan JE, et al. A(3) adenosine receptor activation attenuates neutrophil function and neutrophil-mediated reperfusion injury. Am J Physiol. 1999;277(5 Pt 2):H1895–H905.PubMedGoogle Scholar
  36. 36.
    Foxman EF, Kunkel EJ, Butcher EC. Integrating conflicting chemotactic signals. The role of memory in leukocyte navigation. J Cell Biol. 1999;147(3):577–88.CrossRefPubMedPubMedCentralGoogle Scholar
  37. 37.
    McLeish KR, et al. Exocytosis of neutrophil granule subsets and activation of prolyl isomerase 1 are required for respiratory burst priming. J Innate Immun. 2013;5(3):277–89.CrossRefPubMedPubMedCentralGoogle Scholar
  38. 38.
    Rolas L, et al. Inhibition of mammalian target of rapamycin aggravates the respiratory burst defect of neutrophils from decompensated patients with cirrhosis. Hepatology. 2013;57(3):1163–71.CrossRefPubMedGoogle Scholar
  39. 39.
    Marin V, et al. The p38 mitogen-activated protein kinase pathway plays a critical role in thrombin-induced endothelial chemokine production and leukocyte recruitment. Blood. 2001;98(3):667–73.CrossRefPubMedGoogle Scholar
  40. 40.
    Pouliot M, et al. Expression and activity of prostaglandin endoperoxide synthase-2 in agonist-activated human neutrophils. FASEB J. 1998;12(12):1109–23.PubMedGoogle Scholar
  41. 41.
    Armstrong RA. Investigation of the inhibitory effects of PGE2 and selective EP agonists on chemotaxis of human neutrophils. Br J Pharmacol. 1995;116(7):2903–8.CrossRefPubMedPubMedCentralGoogle Scholar
  42. 42.
    Flamand N, et al. Cyclic AMP-mediated inhibition of 5-lipoxygenase translocation and leukotriene biosynthesis in human neutrophils. Mol Pharmacol. 2002;62(2):250–6.CrossRefPubMedGoogle Scholar
  43. 43.
    Flamand N, et al. Adenosine, a potent natural suppressor of arachidonic acid release and leukotriene biosynthesis in human neutrophils. Am J Respir Crit Care Med. 2000;161(2 Pt 2):S88–S94.CrossRefPubMedGoogle Scholar
  44. 44.
    Cronstein BN, et al. The adenosine/neutrophil paradox resolved: human neutrophils possess both A1 and A2 receptors that promote chemotaxis and inhibit O2 generation, respectively. J Clin Invest. 1990;85(4):1150–7.CrossRefPubMedPubMedCentralGoogle Scholar
  45. 45.
    Cadieux JS, et al. Potentiation of neutrophil cyclooxygenase-2 by adenosine: an early anti-inflammatory signal. J Cell Sci. 2005;118(Pt 7):1437–47.CrossRefPubMedPubMedCentralGoogle Scholar
  46. 46.
    Pouliot M, et al. Adenosine up-regulates cyclooxygenase-2 in human granulocytes: impact on the balance of eicosanoid generation. J Immunol. 2002;169(9):5279–86.CrossRefPubMedGoogle Scholar

Copyright information

© Springer International Publishing 2017

Authors and Affiliations

  • Xiaohan Xu
    • 1
  • Shuyun Zheng
    • 2
  • Yuyun Xiong
    • 3
  • Xu Wang
    • 1
  • Weiting Qin
    • 1
  • Huafeng Zhang
    • 1
  • Bingwei Sun
    • 1
    Email author
  1. 1.Department of Burn and Plastic Surgery, Affiliated HospitalJiangsu UniversityZhenjiangChina
  2. 2.Department of Critical Care Medicine, Nanjing First HospitalNanjing Medical UniversityNanjingChina
  3. 3.Department of Clinical Laboratory, Affiliated HospitalJiangsu UniversityZhenjiangChina

Personalised recommendations