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  • 1
    Language: English
    In: mBio, 10 October 2017, Vol.8(5)
    Description: is the causative agent of tularemia and a potential bioterrorism agent. In the present study, we isolated, identified, and quantified the proteins present in the membranes of the virulent type A strain, Schu S4, and the attenuated type B strain, LVS (live vaccine strain). Spectral counting of mass spectrometric data showed enrichment for membrane proteins in both strains. Mice vaccinated with whole LVS membranes encapsulated in poly (lactic-co-glycolic acid) (PLGA) nanoparticles containing the adjuvant polyinosinic-polycytidylic acid [poly(I·C)] showed significant protection against a challenge with LVS compared to the results seen with naive mice or mice vaccinated with either membranes or poly(I·C) alone. The PLGA-encapsulated Schu S4 membranes with poly(I·C) alone did not significantly protect mice from a lethal intraperitoneal challenge with Schu S4; however, this vaccination strategy provided protection from LVS challenge. Mice that received the encapsulated Schu S4 membranes followed by a booster of LVS bacteria showed significant protection with respect to a lethal Schu S4 challenge compared to control mice. Western blot analyses of the sera from the Schu S4-vaccinated mice that received an LVS booster showed four immunoreactive bands. One of these bands from the corresponding one-dimensional (1D) SDS-PAGE experiment represented capsule. The remaining bands were excised, digested with trypsin, and analyzed using mass spectrometry. The most abundant proteins present in these immunoreactive samples were an outer membrane OmpA-like protein, FopA; the type IV pilus fiber building block protein; a hypothetical membrane protein; and lipoproteins LpnA and Lpp3. These proteins should serve as potential targets for future recombinant protein vaccination studies. The low infectious dose, the high potential mortality/morbidity rates, and the ability to be disseminated as an aerosol make a potential agent for bioterrorism. These characteristics led the Centers for Disease Control (CDC) to classify as a Tier 1 pathogen. Currently, there is no vaccine approved for general use in the United States.
    Keywords: Franscisella Tularensis ; Immunity ; Inner Membrane Proteins ; Outer Membrane Proteins ; Proteomics ; Bacterial Vaccines -- Immunology ; Francisella Tularensis -- Immunology ; Membrane Proteins -- Immunology ; Tularemia -- Prevention & Control ; Vaccines, Subunit -- Immunology
    E-ISSN: 2150-7511
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  • 2
    Language: English
    In: Infection and immunity, 04 January 2016, Vol.84(3), pp.765-74
    Description: Haemophilus haemolyticus and nontypeable Haemophilus influenzae (NTHi) are closely related upper airway commensal bacteria that are difficult to distinguish phenotypically. NTHi causes upper and lower airway tract infections in individuals with compromised airways, while H. haemolyticus rarely causes such infections. The lipooligosaccharide (LOS) is an outer membrane component of both species and plays a role in NTHi pathogenesis. In this study, comparative analyses of the LOS structures and corresponding biosynthesis genes were performed. Mass spectrometric and immunochemical analyses showed that NTHi LOS contained terminal sialic acid more frequently and to a higher extent than H. haemolyticus LOS did. Genomic analyses of 10 strains demonstrated that H. haemolyticus lacked the sialyltransferase genes lic3A and lic3B (9/10) and siaA (10/10), but all strains contained the sialic acid uptake genes siaP and siaT (10/10). However, isothermal titration calorimetry analyses of SiaP from two H. haemolyticus strains showed a 3.4- to 7.3-fold lower affinity for sialic acid compared to that of NTHi SiaP. Additionally, mass spectrometric and immunochemical analyses showed that the LOS from H. haemolyticus contained phosphorylcholine (ChoP) less frequently than the LOS from NTHi strains. These differences observed in the levels of sialic acid and ChoP incorporation in the LOS structures from H. haemolyticus and NTHi may explain some of the differences in their propensities to cause disease.
    Keywords: Haemophilus -- Metabolism ; Haemophilus Infections -- Microbiology ; Haemophilus Influenzae -- Metabolism ; Lipopolysaccharides -- Chemistry ; N-Acetylneuraminic Acid -- Analysis ; Phosphorylcholine -- Analysis
    ISSN: 00199567
    E-ISSN: 1098-5522
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  • 3
    Language: English
    In: Biomedical & Environmental Mass Spectrometry, November 1990, Vol.19(11), pp.731-745
    Description: A structural model is proposed for the surface glycolipids, or lipooligosaccharides (LOS), of gram‐negative pathogenic bacteria that colonize human mucosae, e.g. and The development of this model has involved analysis of a series of pyocin‐resistant mutants with altered LOS and other recent immunochemical and structural data. A comprehensive approach to determining the necessary structural data has been constructed that utilizes liquid secondary ion mass spectrometry, tandem mass spectrometry, methylation analysis and nuclear magnetic resonance. To prepare purified oligosaccharides for these analyses, chromatographic and chemical techniques have been developed that include high‐pH anion‐exchange chromatography of underivatized oligosaccharides and reverse‐phase chromatography after derivatization with hydrazino alkyl benzoates. The proposed LOS model has several unique features that distinguish it from models developed for the lipopolysaccharides of enteric bacteria. This information should lead to an understanding of the unique structure/function relationship of LOS and to the development of carbohydrate‐based vaccines.
    Keywords: Bakterien ; Krankheitserreger ; Lipoprotein ; Strukturanalyse ; Oberflaechenstruktur ; Immunozytochemie ; Massenspektrometrie ; Kernresonanzspektrometrie ; Ionenaustauschchromatographie ; Zellmembran ; Theoretisches Modell ; Oligosaccharid ; Biology ; Anatomy & Physiology;
    ISSN: 0887-6134
    E-ISSN: 1096-9888
    E-ISSN: 23763868
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