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DOI: 10.1055/s-0039-1685495
The Role of Human Platelet Preparation for Toll-Like Receptors 2 and 4 Related Platelet Responsiveness
Funding This publication was funded by the German Research Foundation (DFG) and the University of Würzburg in the funding program Open Access Publishing. Juergen Koessler (project number KO 5256/3–1) and Anna Kobsar (project number KO 5294/2–1) have received grants from Deutsche Forschungsgemeinschaft (DFG).
Publication History
12 November 2018
25 February 2019
Publication Date:
17 April 2019 (online)
Abstract
Background Like immune cells, platelets express the repertoire of toll-like receptors (TLR), among them TLR2 and TLR4, which are important for the recognition of bacterial patterns. Receptor-mediated functional effects in platelets have been investigated, but reliable conclusions are tampered due to heterogeneous study designs with variable platelet preparation methods. This study compares TLR2- and TLR4-dependent platelet responsiveness in platelet-rich plasma (PRP) and in washed platelets (WPs).
Material and Methods Fresh peripheral blood samples from healthy donors served for the preparation of PRP and WP. Basal and agonist-stimulated TLR2 and TLR4 expression levels were evaluated by flow cytometry. Light transmission aggregometry was used to investigate functional effects of TLR2 and TLR4 stimulation with Pam3CSK4 or LPS (lipopolysaccharides from Escherichia coli) as ligands. The capacity of chemokine release was determined by immunoassays.
Results Pam3CSK4 and LPS (in combination with thrombin) were able to induce aggregation in WP, but not in PRP, with threshold concentrations of 15 µg/mL. Basal expression levels of TLR2 and TLR4 were higher in WP than in PRP, increasing several-fold rapidly and persistently upon platelet activation with potent agonists. Pam3CSK4 (15 µg/mL) or LPS led to the submaximal release of RANTES, PF4, PDGF, NAP-2, and sCD40L from WP. In PRP, secretory effects are less pronounced for RANTES, PDGF, or PF4, and not detectable for NAP-2 or sCD40L.
Conclusion The effects mediated by TLR2 and TLR4 stimulation are dependent on platelet preparation, an important issue for experimental designs and for manufacturing of platelet concentrates in transfusion medicine.
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References
- 1 Cognasse F, Nguyen KA, Damien P. , et al. The inflammatory role of platelets via their TLRs and Siglec receptors. Front Immunol 2015; 6: 83
- 2 Morrell CN, Aggrey AA, Chapman LM, Modjeski KL. Emerging roles for platelets as immune and inflammatory cells. Blood 2014; 123 (18) 2759-2767
- 3 von Hundelshausen P, Weber C. Platelets as immune cells: bridging inflammation and cardiovascular disease. Circ Res 2007; 100 (01) 27-40
- 4 Cognasse F, Hamzeh H, Chavarin P, Acquart S, Genin C, Garraud O. Evidence of Toll-like receptor molecules on human platelets. Immunol Cell Biol 2005; 83 (02) 196-198
- 5 Shiraki R, Inoue N, Kawasaki S. , et al. Expression of Toll-like receptors on human platelets. Thromb Res 2004; 113 (06) 379-385
- 6 Hamzeh-Cognasse H, Damien P, Chabert A, Pozzetto B, Cognasse F, Garraud O. Platelets and infections - complex interactions with bacteria. Front Immunol 2015; 6: 82
- 7 Blair P, Rex S, Vitseva O. , et al. Stimulation of toll-like receptor 2 in human platelets induces a thromboinflammatory response through activation of phosphoinositide 3-kinase. Circ Res 2009; 104 (03) 346-354
- 8 Rex S, Beaulieu LM, Perlman DH. , et al. Immune versus thrombotic stimulation of platelets differentially regulates signalling pathways, intracellular protein-protein interactions, and alpha-granule release. Thromb Haemost 2009; 102 (01) 97-110
- 9 Kälvegren H, Skoglund C, Helldahl C, Lerm M, Grenegård M, Bengtsson T. Toll-like receptor 2 stimulation of platelets is mediated by purinergic P2 × 1-dependent Ca2+ mobilisation, cyclooxygenase and purinergic P2Y1 and P2Y12 receptor activation. Thromb Haemost 2010; 103 (02) 398-407
- 10 Aslam R, Speck ER, Kim M. , et al. Platelet Toll-like receptor expression modulates lipopolysaccharide-induced thrombocytopenia and tumor necrosis factor-alpha production in vivo. Blood 2006; 107 (02) 637-641
- 11 Damien P, Cognasse F, Payrastre B. , et al. NF-κB links TLR2 and PAR1 to soluble immunomodulator factor secretion in human platelets. Front Immunol 2017; 8: 85
- 12 Ståhl AL, Svensson M, Mörgelin M. , et al. Lipopolysaccharide from enterohemorrhagic Escherichia coli binds to platelets through TLR4 and CD62 and is detected on circulating platelets in patients with hemolytic uremic syndrome. Blood 2006; 108 (01) 167-176
- 13 Clark SR, Ma AC, Tavener SA. , et al. Platelet TLR4 activates neutrophil extracellular traps to ensnare bacteria in septic blood. Nat Med 2007; 13 (04) 463-469
- 14 Claushuis TAM, Van Der Veen AIP, Horn J. , et al. Platelet Toll-like receptor expression and activation induced by lipopolysaccharide and sepsis. Platelets 2018; :1–9 [Epub ahead of print]
- 15 Rivadeneyra L, Carestia A, Etulain J. , et al. Regulation of platelet responses triggered by Toll-like receptor 2 and 4 ligands is another non-genomic role of nuclear factor-kappaB. Thromb Res 2014; 133 (02) 235-243
- 16 Ward JR, Bingle L, Judge HM. , et al. Agonists of toll-like receptor (TLR)2 and TLR4 are unable to modulate platelet activation by adenosine diphosphate and platelet activating factor. Thromb Haemost 2005; 94 (04) 831-838
- 17 Klarström Engström K, Brommesson C, Kälvegren H, Bengtsson T. Toll like receptor 2/1 mediated platelet adhesion and activation on bacterial mimetic surfaces is dependent on src/Syk-signaling and purinergic receptor P2 × 1 and P2Y12 activation. Biointerphases 2014; 9 (04) 041003
- 18 Berthet J, Damien P, Hamzeh-Cognasse H. , et al. Human platelets can discriminate between various bacterial LPS isoforms via TLR4 signaling and differential cytokine secretion. Clin Immunol 2012; 145 (03) 189-200
- 19 McCabe White M, Jennings LK. Platelet Protocols: Research and Clinical Laboratory Procedures. San Diego, CA: Academic Press; 1999
- 20 Flo TH, Halaas O, Lien E. , et al. Human toll-like receptor 2 mediates monocyte activation by Listeria monocytogenes, but not by group B streptococci or lipopolysaccharide. J Immunol 2000; 164 (04) 2064-2069
- 21 Tsukamoto H, Ukai I, Yamagata Y. , et al. Leucine-rich repeat 2 of human Toll-like receptor 4 contains the binding site for inhibitory monoclonal antibodies. FEBS Lett 2015; 589 (24, Pt B): 3893-3898
- 22 Lievens D, Zernecke A, Seijkens T. , et al. Platelet CD40L mediates thrombotic and inflammatory processes in atherosclerosis. Blood 2010; 116 (20) 4317-4327
- 23 Kato H, Adachi S, Doi T. , et al. Mechanism of collagen-induced release of 5-HT, PDGF-AB and sCD40L from human platelets: role of HSP27 phosphorylation via p44/p42 MAPK. Thromb Res 2010; 126 (01) 39-43
- 24 Brandt E, Petersen F, Ludwig A, Ehlert JE, Bock L, Flad HD. The beta-thromboglobulins and platelet factor 4: blood platelet-derived CXC chemokines with divergent roles in early neutrophil regulation. J Leukoc Biol 2000; 67 (04) 471-478
- 25 von Hundelshausen P, Koenen RR, Sack M. , et al. Heterophilic interactions of platelet factor 4 and RANTES promote monocyte arrest on endothelium. Blood 2005; 105 (03) 924-930
- 26 Fleischer J, Grage-Griebenow E, Kasper B. , et al. Platelet factor 4 inhibits proliferation and cytokine release of activated human T cells. J Immunol 2002; 169 (02) 770-777
- 27 Brown AJ, Sepuru KM, Sawant KV, Rajarathnam K. Platelet-derived chemokine CXCL7 dimer preferentially exists in the glycosaminoglycan-bound form: implications for neutrophil-platelet crosstalk. Front Immunol 2017; 8: 1248
- 28 Antczak AJ, Singh N, Gay SR, Worth RG. IgG-complex stimulated platelets: a source of sCD40L and RANTES in initiation of inflammatory cascade. Cell Immunol 2010; 263 (01) 129-133
- 29 Anselmo A, Riva F, Gentile S. , et al. Expression and function of IL-1R8 (TIR8/SIGIRR): a regulatory member of the IL-1 receptor family in platelets. Cardiovasc Res 2016; 111 (04) 373-384
- 30 Tsai JC, Lin YW, Huang CY. , et al. The role of calpain-myosin 9-Rab7b pathway in mediating the expression of Toll-like receptor 4 in platelets: a novel mechanism involved in α-granules trafficking. PLoS One 2014; 9 (01) e85833
- 31 Duerschmied D, Bode C, Ahrens I. Immune functions of platelets. Thromb Haemost 2014; 112 (04) 678-691
- 32 Semple JW, Italiano Jr JE, Freedman J. Platelets and the immune continuum. Nat Rev Immunol 2011; 11 (04) 264-274
- 33 Aloui C, Prigent A, Sut C. , et al. The signaling role of CD40 ligand in platelet biology and in platelet component transfusion. Int J Mol Sci 2014; 15 (12) 22342-22364
- 34 Gerskowitch VP, Hodge J, Hull RA. , et al. Unexpected relationship between plasma protein binding and the pharmacodynamics of 2-NAP, a CCK1-receptor antagonist. Br J Clin Pharmacol 2007; 63 (05) 618-622
- 35 Koessler J, Hermann S, Weber K. , et al. Role of purinergic receptor expression and function for reduced responsiveness to adenosine diphosphate in washed human platelets. PLoS One 2016; 11 (01) e0147370
- 36 Koessler J, Weber K, Koessler A, Yilmaz P, Boeck M, Kobsar A. Expression and function of purinergic receptors in platelets from apheresis-derived platelet concentrates. Blood Transfus 2016; 14 (06) 545-551
- 37 Jonnalagadda D, Izu LT, Whiteheart SW. Platelet secretion is kinetically heterogeneous in an agonist-responsive manner. Blood 2012; 120 (26) 5209-5216
- 38 Kaufman J, Spinelli SL, Schultz E, Blumberg N, Phipps RP. Release of biologically active CD154 during collection and storage of platelet concentrates prepared for transfusion. J Thromb Haemost 2007; 5 (04) 788-796