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Generation of platelet-derived microparticles through the activation of the toll-like receptor 4

M Alarcón1

  • 1Thrombosis Research Center, Department of Clinical Biochemistry and Immunohematology, Faculty of Health Sciences, Research Center for Aging, Universidad de Talca, 2 Norte 685, Talca, Post code 3460000, Chile.

Heliyon
|April 23, 2019
PubMed
Abstract

Insights

Bacterial infections activate human platelets via toll-like receptor 4 (TLR4), increasing pro-thrombotic Platelet-derived Microparticles (PDMPs). A new protocol efficiently generates and measures PDMPs, aiding cardiovascular disease research.

Area of Science:

  • Immunology
  • Hematology
  • Cardiovascular Research

Background:

  • Bacterial infections can elevate thrombosis and atherosclerosis risk through proinflammatory factors.
  • Human platelets express toll-like receptor 4 (TLR4), a key receptor for bacterial lipopolysaccharide (LPS).
  • Platelet activation generates abundant Platelet-derived Microparticles (PDMPs), linked to Cardiovascular Diseases (CVDs).

Purpose of the Study:

  • To establish a protocol for generating pro-thrombotic microparticles from activated platelets in vitro.
  • To evaluate the role of TLR4 activation in Platelet-derived Microparticles (PDMPs) generation.
  • To assess the potential for inhibiting PDMPs formation.

Main Methods:

  • Washed platelets from healthy volunteers were incubated with Escherichia coli LPS and ADP.
  • Platelet-rich plasma and washed platelets were compared as sources for microparticle generation.
  • Flow cytometry was used to analyze Platelet-derived Microparticles (PDMPs) following TLR4 activation and inhibition assays.

Main Results:

  • Incubation of platelets with LPS and ADP significantly increased Platelet-derived Microparticles (PDMPs) generation by 34-fold.
  • Washed platelets proved to be a superior source for microparticle generation compared to platelet-rich plasma.
  • Acetylsalicylic acid demonstrated a 7.7-fold inhibition of Platelet-derived Microparticles (PDMPs) generation induced by TLR4 activation.

Conclusions:

  • A novel, straightforward protocol for generating and analyzing Platelet-derived Microparticles (PDMPs) using flow cytometry has been developed.
  • This protocol offers a potential method for future studies investigating the association of PDMPs with various pathologies.
  • The findings highlight the role of TLR4 in platelet activation and PDMPs production, with implications for cardiovascular research.
Keywords:
BiochemistryImmunology

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