title: 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: Institute of Medical Microbiology, University of Duisburg-Essen, 45147 Essen, Germany.
authors: Chiranjeevi
authors: C
authors: Chebrolu
authors: Department of Chemistry, University of Natural Resources and Life Sciences, 1190 Vienna, Austria.
authors: Daniel
authors: D
authors: Artner
authors: Institute of Medical Microbiology, University of Duisburg-Essen, 45147 Essen, Germany.
authors: Anna M
authors: AM
authors: Sigmund
authors: Institute of Medical Microbiology, University of Duisburg-Essen, 45147 Essen, Germany.
authors: Jan
authors: J
authors: Buer
authors: Department of Chemistry, University of Natural Resources and Life Sciences, 1190 Vienna, Austria.
authors: Alla
authors: A
authors: Zamyatina
authors: Institute of Medical Microbiology, University of Duisburg-Essen, 45147 Essen, Germany. Electronic address: carsten.kirschning@uk-essen.de.
authors: Carsten J
authors: CJ
authors: Kirschning
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000378:metabolism; D004187:Disaccharides / Q000737:chemistry; D005947:Glucose / Q000737:chemistry*; D057809:HEK293 Cells; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000187:drug effects / Q000378:metabolism*; D008050:Lipid A / Q000737:chemistry* / Q000494:pharmacology*; D051379:Mice; D018716:Molecular Mimicry; D013045:Species Specificity; D051197:Toll-Like Receptor 4 / Q000037:antagonists & inhibitors* / Q000378:metabolism
chemical_list: D004187:Disaccharides; D008050:Lipid A; D051197:Toll-Like Receptor 4; D005947:Glucose
keywords: CD14; LPS; MD-2; PRRs; TLR4; Therapeutic blockade