title: Brucella species release a specific, protease-sensitive, inhibitor of TNF-alpha expression, active on human macrophage-like cells.
abstract: Brucella species can establish themselves and cause disease in humans, but the mechanisms by which brucellae evade the antibacterial defenses of their host remain largely unknown. We have previously reported that, unlike Escherichia coli K12, intracellular pathogens from the genus Brucella survive and multiply within U937-derived phagocytes, and live Brucella organisms failed to induce TNF-alpha release upon infection. Moreover, exogenously added TNF-alpha restricted intracellular growth of Brucella species. Herein, we demonstrate that Brucella-infected U937 cells are activated to express IL-1 beta and IL-6 at both the mRNA and protein levels, while they cannot accumulate TNF-alpha mRNA. When physically separated from macrophages, live brucellae impaired TNF-alpha production in E. coli-infected cells. Moreover, in agonist-activated macrophages, supernatants from Brucella cultures promoted an inhibition of the induction of both TNF-alpha expression and release, without affecting IL-1 beta or IL-6 induction. These phenomena, observed whatever the Brucella strain assayed, show that brucellae release some high m.w. factor(s) that specifically inhibits TNF-alpha expression in activated human macrophages. The proteic nature of the factor(s) was demonstrated by its heat and protease sensitiveness, and this could explain why U937-derived macrophages did release TNF-alpha when infected with chloramphenicol-treated brucellae. We also found that the Brucella factor(s) specifically acts on human macrophagic cells, but not on murine macrophage-like cells. Our findings provide direct evidence that a secreted Brucella virulence factor(s) inhibiting TNF-alpha expression might contribute to the evasion of Brucella organisms from human antimicrobial defenses.
authors.affiliation: INSERM Unit 431, University of Montpellier II, France.
authors.forename: E
authors.initials: E
authors.lastname: Caron
authors.forename: A
authors.initials: A
authors.lastname: Gross
authors.forename: J P
authors.initials: JP
authors.lastname: Liautard
authors.forename: J
authors.initials: J
authors.lastname: Dornand
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000276:immunology* / Q000494:pharmacology*; D001483:Base Sequence; D002002:Brucella / Q000276:immunology* / Q000378:metabolism; D002460:Cell Line; D002474:Cell-Free System / Q000276:immunology; D003470:Culture Media; D010450:Endopeptidases / Q000494:pharmacology*; D006801:Humans; D008262:Macrophage Activation / Q000276:immunology; D008264:Macrophages / Q000201:enzymology / Q000276:immunology* / Q000382:microbiology*; D051379:Mice; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000032:analysis; D013045:Species Specificity; D014409:Tumor Necrosis Factor-alpha / Q000037:antagonists & inhibitors* / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D003470:Culture Media; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; D010450:Endopeptidases