Non-Anticoagulant Fractions of Enoxaparin Suppress Inflammatory Cytokine Release from Peripheral Blood Mononuclear Cells of Allergic Asthmatic Individuals

PLoS One. 2015 Jun 5;10(6):e0128803. doi: 10.1371/journal.pone.0128803. eCollection 2015.

Abstract

Background: Enoxaparin, a low-molecular-weight heparin, is known to possess anti-inflammatory properties. However, its clinical exploitation as an anti-inflammatory agent is hampered by its anticoagulant effect and the associated risk of bleeding.

Objective: The aim of the current study was to examine the ability of non-anticoagulant fractions of enoxaparin to inhibit the release of key inflammatory cytokines in primed peripheral blood mononuclear cells derived from allergic mild asthmatics.

Methods: Peripheral blood mononuclear cells from allergic asthmatics were activated with phytohaemag glutinin (PHA), concanavalin-A (ConA) or phorbol 12-myristate 13-acetate (PMA) in the presence or absence of enoxaparin fractions before cytokine levels were quantified using specific cytokine bead arrays. Together with nuclear magnetic resonance analysis,time-dependent and target-specific effects of enoxaparin fractions were used to elucidate structural determinants for their anti-inflammatory effect and gain mechanistic insights into their anti-inflammatory activity.

Results: Two non-anticoagulant fractions of enoxaparin were identified that significantly inhibited T-cell activation. A disaccharide fraction of enoxaparin inhibited the release of IL-4, IL-5, IL-13 and TNF-α by more than 57% while a tetrasaccharide fraction was found to inhibit the release of tested cytokines by more than 68%. Our data suggest that the observed response is likely to be due to an interaction of 6-O-sulfated tetrasaccharide with cellular receptor(s).

Conclusion and clinical relevance: The two identified anti-inflammatory fractions lacked anticoagulant activity and are therefore not associated with risk of bleeding. The findings highlight the potential therapeutic use of enoxaparin-derived fractions, in particular tetrasaccharide, in patients with chronic inflammatory disorders.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Anti-Inflammatory Agents, Non-Steroidal / isolation & purification
  • Anti-Inflammatory Agents, Non-Steroidal / pharmacology*
  • Asthma / immunology
  • Asthma / pathology
  • Chemical Fractionation
  • Concanavalin A / pharmacology
  • Enoxaparin / chemistry*
  • Female
  • Humans
  • Interleukin-13 / antagonists & inhibitors
  • Interleukin-13 / biosynthesis
  • Interleukin-4 / antagonists & inhibitors
  • Interleukin-4 / biosynthesis
  • Interleukin-5 / antagonists & inhibitors
  • Interleukin-5 / biosynthesis
  • Leukocytes, Mononuclear / drug effects*
  • Leukocytes, Mononuclear / immunology
  • Leukocytes, Mononuclear / pathology
  • Male
  • Middle Aged
  • Oligosaccharides / isolation & purification
  • Oligosaccharides / pharmacology*
  • Phytohemagglutinins / pharmacology
  • Primary Cell Culture
  • Tetradecanoylphorbol Acetate / pharmacology
  • Tumor Necrosis Factor-alpha / antagonists & inhibitors
  • Tumor Necrosis Factor-alpha / biosynthesis

Substances

  • Anti-Inflammatory Agents, Non-Steroidal
  • Enoxaparin
  • IL13 protein, human
  • IL4 protein, human
  • IL5 protein, human
  • Interleukin-13
  • Interleukin-5
  • Oligosaccharides
  • Phytohemagglutinins
  • Tumor Necrosis Factor-alpha
  • Concanavalin A
  • Interleukin-4
  • Tetradecanoylphorbol Acetate

Grants and funding

This work was supported by the Starter Grant from Royal Hobart Hospital Research Foundation, Australia (Grant Number- H22074) awarded to NS, RP and MS (http://www.rhhresearchfoundation.org/index.php/site/research/), Research Enhancement Grant Schemes from University of Tasmania, Australia (Grant Number- 105557) awarded to RP and NG (http://www.utas.edu.au/research-admin/funding/funding-options/internal-funding). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.