title: Activation of human THP-1 cells and rat bone marrow-derived macrophages by Helicobacter pylori lipopolysaccharide.
abstract: The mechanism by which Helicobacter pylori, which has little or no invasive activity, induces gastric-tissue inflammation and injury has not been well characterized. We have previously demonstrated that water-extracted proteins of H. pylori are capable of activating human monocytes by a lipopolysaccharide (LPS)-independent mechanism. We have now compared activation of macrophages by purified LPS from H. pylori and from Escherichia coli. LPS was prepared by phenol-water extraction from H. pylori 88-23 and from E. coli O55. THP-1, a human promyelomonocytic cell line, and macrophages derived from rat bone marrow each were incubated with the LPS preparations, and cell culture supernatants were assayed for production of tumor necrosis factor alpha (TNF-alpha), prostaglandin E2 (PGE2), and nitric oxide. THP-1 cells showed maximal activation by the LPS molecules after cell differentiation was induced by phorbol 12-myristate 13-acetate. Maximal TNF-alpha and PGE2 production occurred by 6 and 18 h, respectively, in both types of cells. In contrast, NO was produced by rat bone marrow-derived macrophages only and was maximal at 18 h. The minimum concentration of purified LPS required to induce TNF-alpha, PGE2, and NO responses in both types of cells was 2,000- to 30,000-fold higher for H. pylori than for E. coli. Purified LPS from three other H. pylori strains with different polysaccharide side chain lengths showed a similarly low level of activity, and polymyxin B treatment markedly reduced activity as well, suggesting that activation was a lipid A phenomenon. These results indicate the low biological activity of H. pylori LPS in mediating macrophage activation.
authors.affiliation: Department of Medicine, Vanderbilt University School of Medicine, Nashville, Tennessee 37232-2605.
authors.forename: G I
authors.initials: GI
authors.lastname: Pérez-Pérez
authors.forename: V L
authors.initials: VL
authors.lastname: Shepherd
authors.forename: J D
authors.initials: JD
authors.lastname: Morrow
authors.forename: M J
authors.initials: MJ
authors.lastname: Blaser
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells; D002454:Cell Differentiation; D002460:Cell Line; D004305:Dose-Response Relationship, Drug; D004926:Escherichia coli; D016480:Helicobacter pylori / Q000276:immunology*; D066298:In Vitro Techniques; D008070:Lipopolysaccharides / Q000276:immunology*; D008262:Macrophage Activation / Q000187:drug effects*; D008264:Macrophages / Q000187:drug effects; D009000:Monocytes / Q000166:cytology / Q000187:drug effects*; D011112:Polymyxin B / Q000494:pharmacology; D051381:Rats; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis
chemical_list: D008070:Lipopolysaccharides; D014409:Tumor Necrosis Factor-alpha; D011112:Polymyxin B