Macrophage tolerance response to Aggregatibacter actinomycetemcomitans lipopolysaccharide induces differential regulation of tumor necrosis factor‐α, interleukin‐1β and matrix metalloproteinase 9 secretion

Abstract

Background and objective The lipopolysaccharide of Aggregatibacter actinomycetemcomitans, a potent stimulator of the immune system, induces the secretion of inflammatory mediators that modulate periodontal tissue destruction. In this study, we investigated the tolerance response of human macrophages to stimulation with A. actinomycetemcomitans lipopolysaccharide. Material and methods U937 monocytes were differentiated into adherent macrophage-like cells by treatment with phorbol myristic acid. Macrophage-like cells were then pretreated for 24 h with either 0.01 or 0.1 microg/mL LPS A. actinomycetemcomitans. Culture medium supernatants were removed and cells were restimulated with LPS at 1 microg/mL. Cell-free supernatants were collected after 24 h of stimulation and analyzed by ELISA for TNF-alpha, IL-1 beta, IL-6, IL-8, PGE(2) and MMP-9. Results Phorbol myristic acid-differentiated U937 macrophages treated with low doses of lipopolysaccharide developed tolerance to subsequent lipopolysaccharide treatments, resulting in significantly reduced secretion of tumor necrosis factor-alpha. However, this tolerance response was associated with increased secretion of interleukin-1 beta and matrix metalloproteinase 9, whereas the secretion of interleukin-6, interleukin-8 and prostaglandin E(2) was unaffected. Phosphatidylinositol-3'-kinase inhibitors added during the tolerance-induction period markedly attenuated the increase in interleukin-1 beta secretion but had no effect on tumor ne…