Species and mediator specific TLR4 antagonism in primary human and murine immune cells by βGlcN(1↔1)αGlc based lipid A mimetics.

Abstract

Immune stimulatory pathogen associated molecular patterns (PAMPs) are major drivers of infection pathology. Infections with Gram-negative bacteria or negatively polar and single stranded RNA influenza virus are prominent causes of morbidity and mortality. Toll-like receptor (TLR) 4 is a major host sensor for both of the two infections. In order to inhibit TLR4 driven immune activation we recently developed synthetic tetra-acylated lipid A mimetics based on a conformationally restricted βGlcN(1↔1)αGlcN disaccharide scaffold (DA-compounds) that antagonized ectopically overexpressed human and murine TLR4/MD-2 complexes. Here we comparatively analyzed human peripheral blood mononuclear cell (hPBMC) and murine bone marrow derived macrophage (mBM) activation upon 30 min of preincubation in vitro with six variably acylated DA-compounds. 16 h subsequent to consequent LPS challenge, we sampled culture supernatants for cytokine and NO concentration analysis. Four compounds significantly inhibited release of both TNF and IL-6 by hPBMCs upon LPS challenge. In contrast, three compounds effectively inhibited mBM production of MIP-2 and KC, and even five of them inhibited IL-6 and NO production. LPS driven like other TLR ligand driven mBM TNF release was largely unimpaired. The inhibitory effect was specific in that Clo75 driven cytokine release by both hPBMCs and mBMs was unimpaired by the compounds analyzed. Our results indicate biological species specificity of LPS antagonism by variably tetraacylated lipid A mimetics and validate three out of six DA-antagonists as promising candidates for development of therapeutically applicable anti-inflammatory compounds.

• Authors: Chiranjeevi Chebrolu, Daniel Artner, Anna M Sigmund, Jan Buer, Alla Zamyatina, Carsten J Kirschning
• Journal: Molecular immunology (Mol Immunol) Vol. 67 Issue 2 Pt B Pg. 636-41 (Oct 2015) ISSN: 1872-9142 [Electronic] England
• PMID: 26319313 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2015 Elsevier Ltd. All rights reserved.

Chemical References

• Disaccharides
• Lipid A
• Toll-Like Receptor 4
• Glucose

Topics

• Animals
• Bone Marrow Cells (metabolism)
• Disaccharides (chemistry)
• Glucose (chemistry)
• HEK293 Cells
• Humans
• Leukocytes, Mononuclear (drug effects, metabolism)
• Lipid A (chemistry, pharmacology)
• Mice
• Molecular Mimicry
• Species Specificity
• Toll-Like Receptor 4 (antagonists & inhibitors, metabolism)