Anthrax toxin blocks priming of neutrophils by lipopolysaccharide and by muramyl dipeptide.

Abstract

We studied the pretreatment of human polymorphonuclear neutrophils (PMN) with purified preparations of the anthrax toxin components.sbd.protective antigen (PA), edema factor (EF), and lethal factor (LF).sbd.and their effects on release of superoxide anion .**GRAPHIC**. after stimulation with the chemotactic peptide N-formylmethionyl-leucyl-phenylalanine (FMLP). PMN isolated in the absence of lipopolysaccharide (LPS) (< 0.1 ng/ml) released only small amounts of .**GRAPHIC**. after FMLP stimulation; pretreatment with anthrax toxin had little effect. The release of .**GRAPHIC**. was increased fivefold by prior treatment with 3 ng/ml LPS for 1 h at 37.degree. C, an effect referred to as priming. PMN were primed to an equivalent extent by treatment with 100 ng/ml N-acetyl-muramyl-L-alanyl-d-isoglutamine (muramyl dipeptide [MDP]). Pretreatment of PMN with anthrax toxin components PA plus EF or PA plus LF inhibited priming by LPS or MDP, as shown by the reduction in the release of .**GRAPHIC**. up to 90% relative to controls not treated with toxin; single toxin components were inactive. The inhibition was markedly reduced when priming with LPS or MDP was carried out before exposure to toxin. .**GRAPHIC**. release after stimulation by phorbol myristate acetate was not increased by priming, and pretreatment with toxin did not inhibit .**GRAPHIC**. release after this stimulus. Evidently, anthrax toxin inhibits the priming that is norally induced in PMN by bacterial products and is necessary for the full expression of antimicrobial effects.