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Toll like receptor 2/1 mediated platelet adhesion and activation on bacterial mimetic surfaces is dependent on src/Syk-signaling and purinergic receptor P2X1 and P2Y12 activation
Örebro University, School of Health and Medical Sciences, Örebro University, Sweden.
Department of Physics, Chemistry and Biology, Division of Molecular Surface Physics and Nanoscience, Linköping University, Linköping, Sweden.
Department of Clinical Pathology and Clinical Genetics, Linköping University Hospital, Linköping, Sweden; Department of Clinical and Experimental Medicine, Linköping University, Linköping, Sweden .
Örebro University, School of Medicine, Örebro University, Sweden.
2014 (English)In: Biointerphases, ISSN 1934-8630, E-ISSN 1559-4106, Vol. 9, no 4, 041003- p.Article in journal (Refereed) Published
Abstract [en]

Platelets are considered to have important functions in inflammatory processes as key players in innate immunity. Toll like receptors (TLRs), expressed on platelets, recognize pathogen associated molecular patterns and trigger immune responses. Pathogens are able to adhere to human tissues and form biofilms which cause a continuous activation of the immune system. The authors aimed to investigate how immobilized Pam(3)CSK(4) (a synthetic TLR2/1 agonist) and IgG, respectively, resembling a bacterial focus, affects adhesion and activation of platelets including release of two cytokines, regulated on activation normal T-cell expressed and secreted (RANTES) and macrophage migration inhibitory factor (MIF). The authors also aim to clarify the signaling downstream of TLR2/1 and Fc gamma RII (IgG receptor) and the role of adenine nucleotides in this process. Biolayers of Pam(3)CSK(4) and IgG, respectively, were confirmed by null-ellipsometry and contact angle measurements. Platelets were preincubated with signaling inhibitors for scr and Syk and antagonists for P2X1 or P2Y1 [adenosine triphosphate (ATP), adenosine diphosphate (ADP) receptors] prior to addition to the surfaces. The authors show that platelets adhere and spread on both Pam(3)CSK(4)- and IgG-coated surfaces and that this process is antagonized by scr and Syc inhibitors as well as P2X1 and P2Y antagonists. This suggests that Pam(3)CSK(4) activated platelets utilize the same pathway as Fc gamma RII. Moreover, the authors show that ATP-ligation of P2X1 is of importance for further platelet activation after TLR2/1-activation, and that P2Y12 is the prominent ADP-receptor involved in adhesion and spreading. RANTES and MIF were secreted over time from platelets adhering to the coated surfaces, but no MIF was released upon stimulation with soluble Pam(3)CSK(4). These results clarify the importance of TLR2/1 and Fc gamma RII in platelet adhesion and activation, and strengthen the role of platelets as an active player in sensing bacterial infections.

Place, publisher, year, edition, pages
2014. Vol. 9, no 4, 041003- p.
National Category
Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)
Research subject
Cell Research
Identifiers
URN: urn:nbn:se:oru:diva-42365DOI: 10.1116/1.4901135ISI: 000347160900004PubMedID: 25553878OAI: oai:DiVA.org:oru-42365DiVA: diva2:785900
Funder
Swedish Heart Lung FoundationSwedish Society for Medical Research (SSMF)
Note

Funding Agencies:

Swedish Medical Research Council

Olle Engkvist Foundation

Lars Hiertas Minne foundation

Available from: 2015-02-04 Created: 2015-02-03 Last updated: 2017-10-18Bibliographically approved
In thesis
1. Platelets as immune cells in sensing bacterial infection
Open this publication in new window or tab >>Platelets as immune cells in sensing bacterial infection
2015 (English)Doctoral thesis, comprehensive summary (Other academic)
Place, publisher, year, edition, pages
Örebro: Örebro university, 2015. 78 p.
Series
Örebro Studies in Medicine, ISSN 1652-4063 ; 116
Keyword
Platelets, infection, TLR2/1, Porphyromonas gingivalis, inflammatory mediators, lipid peroxidation
National Category
Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)
Research subject
Cell Research
Identifiers
urn:nbn:se:oru:diva-40397 (URN)978-91-7529-060-7 (ISBN)
Public defence
2015-03-06, Universitetssjukhuset, Wilandersalen, Södra Grev Rosengatan, Örebro, 09:00 (Swedish)
Opponent
Supervisors
Available from: 2015-01-08 Created: 2015-01-08 Last updated: 2017-10-17Bibliographically approved

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School of Health and Medical Sciences, Örebro University, SwedenSchool of Medicine, Örebro University, Sweden
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