Skip to Content
Merck
  • Human filarial Wolbachia lipopeptide directly activates human neutrophils in vitro.

Human filarial Wolbachia lipopeptide directly activates human neutrophils in vitro.

Parasite immunology (2014-06-10)
F Tamarozzi, H L Wright, K L Johnston, S W Edwards, J D Turner, M J Taylor
ABSTRACT

The host inflammatory response to the Onchocerca volvulus endosymbiont, Wolbachia, is a major contributing factor in the development of chronic pathology in humans (onchocerciasis/river blindness). Recently, the toll-like pattern recognition receptor motif of the major inflammatory ligands of filarial Wolbachia, membrane-associated diacylated lipoproteins, was functionally defined in murine models of pathology, including mediation of neutrophil recruitment to the cornea. However, the extent to which human neutrophils can be activated in response to this Wolbachia pattern recognition motif is not known. Therefore, the responses of purified peripheral blood human neutrophils to a synthetic N-terminal diacylated lipopeptide (WoLP) of filarial Wolbachia peptidoglycan-associated lipoprotein (PAL) were characterized. WoLP exposure led to a dose-dependent activation of healthy, human neutrophils that included gross morphological alterations and modulation of surface expressed integrins involved in tethering, rolling and extravasation. WoLP exposure induced chemotaxis but not chemokinesis of neutrophils, and secretion of the major neutrophil chemokine, interleukin 8. WoLP also induced and primed the respiratory burst, and enhanced neutrophil survival by delay of apoptosis. These results indicate that the major inflammatory motif of filarial Wolbachia lipoproteins directly activates human neutrophils in vitro and promotes a molecular pathway by which human neutrophils are recruited to sites of Onchocerca parasitism.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Ammonium chloride, 99.99% trace metals basis
Sigma-Aldrich
Phorbol 12-myristate 13-acetate, ≥99% (TLC), film or powder
Sigma-Aldrich
Ammonium chloride, Molecular Biology, suitable for cell culture, ≥99.5%
Sigma-Aldrich
Propidium iodide, ≥94.0% (HPLC)
Sigma-Aldrich
Trypan Blue, powder, BioReagent, suitable for cell culture
Sigma-Aldrich
Trypan Blue, ≥80% (HPLC), Dye content 60 %
Sigma-Aldrich
Phorbol 12-myristate 13-acetate, synthetic, ≥98.0% (TLC)
Sigma-Aldrich
Ammonium chloride, tested according to Ph. Eur.
Sigma-Aldrich
Propidium iodide, ≥94% (HPLC)
Sigma-Aldrich
Ammonium chloride, 99.998% trace metals basis
Sigma-Aldrich
PMA, for use in molecular biology applications, ≥99% (HPLC), Molecular Biology
Sigma-Aldrich
Ammonium chloride, BioUltra, ≥99.5% (AT)
Sigma-Aldrich
TNF-α human, Animal-component free, recombinant, expressed in E. coli, suitable for cell culture
Supelco
Ammonium chloride, Pharmaceutical Secondary Standard; Certified Reference Material
Methionine, European Pharmacopoeia (EP) Reference Standard
Dimethyl sulfoxide, European Pharmacopoeia (EP) Reference Standard
USP
Dimethyl sulfoxide, United States Pharmacopeia (USP) Reference Standard
DL-Methionine, European Pharmacopoeia (EP) Reference Standard
Sigma-Aldrich
Dimethyl sulfoxide, suitable for HPLC, ≥99.7%
Sigma-Aldrich
Dimethyl sulfoxide, ReagentPlus®, ≥99.5%
Sigma-Aldrich
Dimethyl sulfoxide, ACS reagent, ≥99.9%
Sigma-Aldrich
Tumor Necrosis Factor-α human, Xeno-free, recombinant, expressed in HEK 293 cells, suitable for cell culture
Phenylalanine, European Pharmacopoeia (EP) Reference Standard
Supelco
Dimethyl sulfoxide, for inorganic trace analysis, ≥99.99995% (metals basis)
Sigma-Aldrich
Trypan Blue solution, 0.4%, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Dimethyl sulfoxide, meets EP testing specifications, meets USP testing specifications
Sigma-Aldrich
Ethylenediaminetetraacetic acid, BioUltra, anhydrous, ≥99% (titration)
Sigma-Aldrich
Ethylenediaminetetraacetic acid, anhydrous, crystalline, BioReagent, suitable for cell culture
Sigma-Aldrich
Tumor Necrosis Factor-α human, TNF-α, recombinant, expressed in E. coli, powder, suitable for cell culture
Sigma-Aldrich
Ethylenediaminetetraacetic acid, purified grade, ≥98.5%, powder