title: Profiles of structural heterogeneity in native lipooligosaccharides of Neisseria and cytokine induction.
abstract: Fine differences in the phosphorylation and acylation of lipooligosaccharide (LOS) from Neisseria species are thought to profoundly influence the virulence of the organisms and the innate immune responses of the host, such as signaling through toll-like receptor 4 (TLR4) and triggering receptor expressed on myeloid cells (TREM). MALDI time-of-flight (TOF) mass spectrometry was used to characterize heterogeneity in the native LOS from Neisseria gonorrheae and N. meningitidis. A sample preparation methodology previously reported for Escherichia coli lipopolysaccharide (LPS) employing deposition of untreated LOS on a thin layer of a film composed of 2,4,6-trihydroxyacetophenone and nitrocellulose was used. Prominent peaks were observed corresponding to molecular ions and to fragment ions primarily formed by cleavage between the 3-deoxy-D-manno-oct-2-ulosonic acid (Kdo) and the lipid A (LA). Analyses of these data and comparison with spectra of the corresponding O-deacylated or hydrogen fluoride-treated LOS enabled the detection of novel species that apparently differed by the expression of up to three phosphates with one or more phosphoethanolamine (PEA) groups on the LA. We found that the heterogeneity profile of acylation and phosphorylation correlates with the induction of proinflammatory cytokines in THP-1 monocytic cells. This methodology enabled us to rapidly profile components of structural variants of native LOS that are of importance biologically.
authors.affiliation: Center for Immunochemistry, Veterans Affairs Medical Center, 4150 Clement Street, San Francisco, CA, 94121.
authors.forename: Constance M
authors.initials: CM
authors.lastname: John
authors.affiliation: Center for Immunochemistry, Veterans Affairs Medical Center, 4150 Clement Street, San Francisco, CA, 94121.
authors.forename: Mingfeng
authors.initials: M
authors.lastname: Liu
authors.affiliation: Center for Immunochemistry, Veterans Affairs Medical Center, 4150 Clement Street, San Francisco, CA, 94121; Department of Laboratory Medicine, University of California, San Francisco, CA 94143. Electronic address: Gary.Jarvis@ucsf.edu.
authors.forename: Gary A
authors.initials: GA
authors.lastname: Jarvis
mesh_terms: D000215:Acylation; D002240:Carbohydrate Sequence; D002460:Cell Line; D016207:Cytokines / Q000096:biosynthesis*; D054884:Host-Pathogen Interactions / Q000276:immunology; D006801:Humans; D006858:Hydrofluoric Acid; D007113:Immunity, Innate; D018836:Inflammation Mediators / Q000378:metabolism; D008070:Lipopolysaccharides / Q000737:chemistry* / Q000276:immunology* / Q000633:toxicity; D008969:Molecular Sequence Data; D015394:Molecular Structure; D009000:Monocytes / Q000187:drug effects / Q000276:immunology / Q000382:microbiology; D009343:Neisseria / Q000737:chemistry* / Q000276:immunology* / Q000472:pathogenicity; D009344:Neisseria gonorrhoeae / Q000737:chemistry / Q000276:immunology / Q000472:pathogenicity; D009345:Neisseria meningitidis / Q000737:chemistry / Q000276:immunology / Q000472:pathogenicity; D010766:Phosphorylation; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014774:Virulence / Q000276:immunology
chemical_list: D016207:Cytokines; D018836:Inflammation Mediators; D008070:Lipopolysaccharides; C023023:lipid-linked oligosaccharides; D006858:Hydrofluoric Acid