Download PDF
Thromb Haemost 2020; 120(04): 658-670
DOI: 10.1055/s-0040-1703007
Blood Cells, Inflammation and Infection
Georg Thieme Verlag KG Stuttgart · New York

Thrombin Promotes Macrophage Polarization into M1-Like Phenotype to Induce Inflammatory Responses

Mercedes López-Zambrano   *
1   Centro de Biofísica y Bioquímica, Instituto Venezolano de Investigaciones Científicas, Caracas, Venezuela
2   Department of Biochemistry, Medical Faculty, Justus-Liebig-University, Giessen, Germany
,
Julián Rodriguez-Montesinos   *
2   Department of Biochemistry, Medical Faculty, Justus-Liebig-University, Giessen, Germany
,
Gustavo E. Crespo-Avilan
2   Department of Biochemistry, Medical Faculty, Justus-Liebig-University, Giessen, Germany
3   Cardiovascular and Metabolic Disorders Program, Duke-NUS Medical School, Singapore
,
Mónica Muñoz-Vega
4   Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubirán, Unidad de Investigación de Enfermedades Metabólicas, Mexico City, Mexico
,
Klaus T. Preissner
2   Department of Biochemistry, Medical Faculty, Justus-Liebig-University, Giessen, Germany
5   Institute of Fundamental Medicine and Biology, Kazan (Volga Region) Federal University, Kazan, Russian Federation
› Author Affiliations
Further Information

Publication History

01 August 2019

24 January 2020

Publication Date:
04 March 2020 (online)

Also available at
    Permissions and Reprints

Abstract

Despite strong evidence supporting the cellular interplay between haemostasis and innate immunity, humoral connections between blood coagulation and the behavior of inflammatory macrophages are not well understood. In this study, we investigated changes in gene expression of selected cytokines and chemokines and their secretion profiles following thrombin stimulation of murine macrophages. Thrombin promoted differentiation of macrophages into an M1-like phenotype that was associated with the expression of classical pro-inflammatory markers. The cellular actions of thrombin on macrophages were mediated in part by protease-activated receptor-1 (PAR-1) and were dependent on phosphoinositide 3-kinase/AKT and nuclear factor-κB. Moreover, heat-denatured thrombin stimulated the secretion of some pro-inflammatory mediators to the same magnitude indicating that different receptors transmit cellular signals of enzymatically active thrombin and nonactive thrombin, the latter remaining so far undefined. Finally, pretreatment of macrophages with thrombin resulted in tolerance against secondary stimulation by lipopolysaccharide with regard to expression of tumor necrosis factor-α. These results provide new insights into the molecular link between the key enzyme of haemostasis and innate immunity responses.

Keywords

thrombin receptors - tumor necrosis factor-α - protease-activated receptors - cytokine profile - innate immunity

Authors' Contributions

M.L.Z. and K.T.P. conceived the overall research design of the present study and wrote the manuscript. M.L.Z. and J.R.M. performed the major experiments as well as the statistical analyses. G.E.C.A. and M.M.V. performed some experiments. All authors approved the submitted version of the manuscript.


* Joint first authors.


Supplementary Material

 

  • References

  • 1 Macfarlane SR, Seatter MJ, Kanke T, Hunter GD, Plevin R. Proteinase-activated receptors. Pharmacol Rev 2001; 53 (02) 245-282
    PubMedGoogle Scholar
  • 2 Davis RP, Miller-Dorey S, Jenne CN. Platelets and coagulation in infection. Clin Transl Immunology 2016; 5 (07) e89
    CrossrefPubMedGoogle Scholar
  • 3 Palta S, Saroa R, Palta A. Overview of the coagulation system. Indian J Anaesth 2014; 58 (05) 515-523
    CrossrefPubMedGoogle Scholar
  • 4 López ML, Bruges G, Crespo G. , et al. Thrombin selectively induces transcription of genes in human monocytes involved in inflammation and wound healing. Thromb Haemost 2014; 112 (05) 992-1001
    Article in Thieme ConnectPubMedGoogle Scholar
  • 5 Coughlin SR. How the protease thrombin talks to cells. Proc Natl Acad Sci U S A 1999; 96 (20) 11023-11027
    CrossrefPubMedGoogle Scholar
  • 6 Ma L, Dorling A. The roles of thrombin and protease-activated receptors in inflammation. Semin Immunopathol 2012; 34 (01) 63-72
    CrossrefPubMedGoogle Scholar
  • 7 Esmon CT, Xu J, Lupu F. Innate immunity and coagulation. J Thromb Haemost 2011; 9 (Suppl. 01) 182-188
    CrossrefPubMedGoogle Scholar
  • 8 Key NS, Vercellotti GM, Winkelmann JC. , et al. Infection of vascular endothelial cells with herpes simplex virus enhances tissue factor activity and reduces thrombomodulin expression. Proc Natl Acad Sci U S A 1990; 87 (18) 7095-7099
    CrossrefPubMedGoogle Scholar
  • 9 Funderburg NT, Mayne E, Sieg SF. , et al. Increased tissue factor expression on circulating monocytes in chronic HIV infection: relationship to in vivo coagulation and immune activation. Blood 2010; 115 (02) 161-167
    CrossrefPubMedGoogle Scholar
  • 10 Veltrop MHAM, Beekhuizen H, Thompson J. Bacterial species- and strain-dependent induction of tissue factor in human vascular endothelial cells. Infect Immun 1999; 67 (11) 6130-6138
    CrossrefPubMedGoogle Scholar
  • 11 Geissmann F, Manz MG, Jung S, Sieweke MH, Merad M, Ley K. Development of monocytes, macrophages, and dendritic cells. Science 2010; 327 (5966): 656-661
    CrossrefPubMedGoogle Scholar
  • 12 Szaba FM, Smiley ST. Roles for thrombin and fibrin(ogen) in cytokine/chemokine production and macrophage adhesion in vivo. Blood 2002; 99 (03) 1053-1059
    CrossrefPubMedGoogle Scholar
  • 13 Zheng L, Martins-Green M. Molecular mechanisms of thrombin-induced interleukin-8 (IL-8/CXCL8) expression in THP-1-derived and primary human macrophages. J Leukoc Biol 2007; 82 (03) 619-629
    CrossrefPubMedGoogle Scholar
  • 14 Pou J, Rebollo A, Piera L. , et al. Tissue factor pathway inhibitor 2 is induced by thrombin in human macrophages. Biochim Biophys Acta 2011; 1813 (06) 1254-1260
    PubMedGoogle Scholar
  • 15 Colognato R, Slupsky JR, Jendrach M, Burysek L, Syrovets T, Simmet T. Differential expression and regulation of protease-activated receptors in human peripheral monocytes and monocyte-derived antigen-presenting cells. Blood 2003; 102 (07) 2645-2652
    CrossrefPubMedGoogle Scholar
  • 16 Cabrera-Fuentes HA, Lopez ML, McCurdy S. , et al. Regulation of monocyte/macrophage polarisation by extracellular RNA. Thromb Haemost 2015; 113 (03) 473-481
    Article in Thieme ConnectPubMedGoogle Scholar
  • 17 Daley JM, Brancato SK, Thomay AA, Reichner JS, Albina JE. The phenotype of murine wound macrophages. J Leukoc Biol 2010; 87 (01) 59-67
    CrossrefPubMedGoogle Scholar
  • 18 Yang Z, Ming XF. Functions of arginase isoforms in macrophage inflammatory responses: impact on cardiovascular diseases and metabolic disorders. Front Immunol 2014; 5: 533
    PubMedGoogle Scholar
  • 19 Bae YS, Yi HJ, Lee HY. , et al. Differential activation of formyl peptide receptor-like 1 by peptide ligands. J Immunol 2003; 171 (12) 6807-6813
    CrossrefPubMedGoogle Scholar
  • 20 Partida-Sánchez S, Randall TD, Lund FE. Innate immunity is regulated by CD38, an ecto-enzyme with ADP-ribosyl cyclase activity. Microbes Infect 2003; 5 (01) 49-58
    CrossrefPubMedGoogle Scholar
  • 21 Amici SA, Young NA, Narvaez-Miranda J. , et al. CD38 Is Robustly Induced in Human macrophages and monocytes in inflammatory conditions. Front Immunol 2018; 9: 1593
    CrossrefPubMedGoogle Scholar
  • 22 Jablonski KA, Amici SA, Webb LM. , et al. Novel markers to delineate murine M1 and M2 macrophages. PLoS One 2015; 10 (12) e0145342
    CrossrefPubMedGoogle Scholar
  • 23 Connolly TM, Condra C, Feng DM. , et al. Species variability in platelet and other cellular responsiveness to thrombin receptor-derived peptides. Thromb Haemost 1994; 72 (04) 627-633
    Article in Thieme ConnectPubMedGoogle Scholar
  • 24 Derian CK, Santulli RJ, Tomko KA, Haertlein BJ, Andrade-Gordon P. Species differences in platelet responses to thrombin and SFLLRN. receptor-mediated calcium mobilization and aggregation, and regulation by protein kinases. Thromb Res 1995; 78 (06) 505-519
    CrossrefPubMedGoogle Scholar
  • 25 Takashiba S, Van Dyke TE, Amar S, Murayama Y, Soskolne AW, Shapira L. Differentiation of monocytes to macrophages primes cells for lipopolysaccharide stimulation via accumulation of cytoplasmic nuclear factor kappaB. Infect Immun 1999; 67 (11) 5573-5578
    CrossrefPubMedGoogle Scholar
  • 26 Rossol M, Heine H, Meusch U. , et al. LPS-induced cytokine production in human monocytes and macrophages. Crit Rev Immunol 2011; 31 (05) 379-446
    CrossrefPubMedGoogle Scholar
  • 27 Parameswaran N, Patial S. Tumor necrosis factor-α signaling in macrophages. Crit Rev Eukaryot Gene Expr 2010; 20 (02) 87-103
    CrossrefPubMedGoogle Scholar
  • 28 Nakanishi-Matsui M, Zheng YW, Sulciner DJ, Weiss EJ, Ludeman MJ, Coughlin SR. PAR3 is a cofactor for PAR4 activation by thrombin. Nature 2000; 404 (6778): 609-613
    CrossrefPubMedGoogle Scholar
  • 29 Mihara K, Ramachandran R, Saifeddine M. , et al. Thrombin-mediated direct activation of proteinase-activated receptor-2: Another target for thrombin signaling. Mol Pharmacol 2016; 89 (05) 606-614
    CrossrefPubMedGoogle Scholar
  • 30 Brummel KE, Paradis SG, Butenas S, Mann KG. Thrombin functions during tissue factor-induced blood coagulation. Blood 2002; 100 (01) 148-152
    CrossrefPubMedGoogle Scholar
  • 31 Preissner KT, Koyama T, Müller D, Tschopp J, Müller-Berghaus G. Domain structure of the endothelial cell receptor thrombomodulin as deduced from modulation of its anticoagulant functions. Evidence for a glycosaminoglycan-dependent secondary binding site for thrombin. J Biol Chem 1990; 265 (09) 4915-4922
    CrossrefPubMedGoogle Scholar
  • 32 Ay C, Dunkler D, Simanek R. , et al. Prediction of venous thromboembolism in patients with cancer by measuring thrombin generation: results from the Vienna Cancer and Thrombosis Study. J Clin Oncol 2011; 29 (15) 2099-2103
    CrossrefPubMedGoogle Scholar
  • 33 Milligan G, Kostenis E. Heterotrimeric G-proteins: a short history. Br J Pharmacol 2006; 147 (Suppl. 01) S46-S55
    CrossrefPubMedGoogle Scholar
  • 34 Cong Y, Oliver AO, Elson CO. Effects of cholera toxin on macrophage production of co-stimulatory cytokines. Eur J Immunol 2001; 31 (01) 64-71
    CrossrefPubMedGoogle Scholar
  • 35 Katada T. The inhibitory G protein G(i) identified as pertussis toxin-catalyzed ADP-ribosylation. Biol Pharm Bull 2012; 35 (12) 2103-2111
    CrossrefPubMedGoogle Scholar
  • 36 Vergadi E, Ieronymaki E, Lyroni K, Vaporidi K, Tsatsanis C. Akt signaling pathway in macrophage activation and M1/M2 polarization. J Immunol 2017; 198 (03) 1006-1014
    CrossrefPubMedGoogle Scholar
  • 37 Bar-Shavit R, Kahn A, Fenton II JW, Wilner GD. Chemotactic response of monocytes to thrombin. J Cell Biol 1983; 96 (01) 282-285
    CrossrefPubMedGoogle Scholar
  • 38 Bar-Shavit R, Benezra M, Eldor A. , et al. Thrombin immobilized to extracellular matrix is a potent mitogen for vascular smooth muscle cells: nonenzymatic mode of action. Cell Regul 1990; 1 (06) 453-463
    CrossrefPubMedGoogle Scholar
  • 39 Bar-Shavit R, Wilner GD. Biologic activities of nonenzymatic thrombin: elucidation of a macrophage interactive domain. Semin Thromb Hemost 1986; 12 (03) 244-249
    Article in Thieme ConnectPubMedGoogle Scholar
  • 40 Tran T, Stewart AG. Protease-activated receptor (PAR)-independent growth and pro-inflammatory actions of thrombin on human cultured airway smooth muscle. Br J Pharmacol 2003; 138 (05) 865-875
    CrossrefPubMedGoogle Scholar
  • 41 Quintin J, Saeed S, Martens JHA. , et al. Candida albicans infection affords protection against reinfection via functional reprogramming of monocytes. Cell Host Microbe 2012; 12 (02) 223-232
    CrossrefPubMedGoogle Scholar
  • 42 Netea MG, Quintin J, van der Meer JW. Trained immunity: a memory for innate host defense. Cell Host Microbe 2011; 9 (05) 355-361
    CrossrefPubMedGoogle Scholar
  • 43 Ifrim DC, Quintin J, Joosten LA. , et al. Trained immunity or tolerance: opposing functional programs induced in human monocytes after engagement of various pattern recognition receptors. Clin Vaccine Immunol 2014; 21 (04) 534-545
    CrossrefPubMedGoogle Scholar