Outer membranes of a lipopolysaccharide-protein complex (LPS-17 kDa protein) as chemical tularemia vaccines

Khlebnikov, V. S.; Golovliov, Igor; Kulevatsky, Dmitrij P.; Tokhtamysheva, Natalia V.; Averin, Sergej F.; Zhemchugov, Vladislav E.; Pchelintsev, S.Yu.; Afanasiev, S. S.; Shcherbakov, Grigorij Ya.

doi:10.1111/j.1574-695x.1996.tb00241.x

Cited by 5 publications

(5 citation statements)

References 14 publications

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“…1 and data not shown). As controls of the purity of the fractions, we detected IglC only in the soluble fraction, as reported previously (50), and the inner membrane protein IcmF/PdpB only in the Sarkosyl-soluble fraction (50), and the outer membrane protein Tul4 (40,69), absent in the soluble fraction, was clearly enriched in the Sarkosyl-insoluble fraction, as expected ( Fig. 1).…”

Section: Resultssupporting

confidence: 57%

IglG and IglI of the Francisella Pathogenicity Island Are Important Virulence Determinants of Francisella tularensis LVS

et al. 2011

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The Gram-negative bacterium Francisella tularensis is the causative agent of tularemia, a disease intimately associated with the multiplication of the bacterium within host macrophages. This in turn requires the expression of Francisella pathogenicity island (FPI) genes, believed to encode a type VI secretion system. While the exact functions of many of the components have yet to be revealed, some have been found to contribute to the ability of Francisella to cause systemic infection in mice as well as to prevent phagolysosomal fusion and facilitate escape into the host cytosol. Upon reaching this compartment, the bacterium rapidly multiplies, inhibits activation of the inflammasome, and ultimately causes apoptosis of the host cell. In this study, we analyzed the contribution of the FPI-encoded proteins IglG, IglI, and PdpE to the aforementioned processes in F. tularensis LVS. The ⌬pdpE mutant behaved similarly to the parental strain in all investigated assays. In contrast, ⌬iglG and ⌬iglI mutants, although they were efficiently replicating in J774A.1 cells, both exhibited delayed phagosomal escape, conferred a delayed activation of the inflammasome, and exhibited reduced cytopathogenicity as well as marked attenuation in the mouse model. Thus, IglG and IglI play key roles for modulation of the intracellular host response and also for the virulence of F. tularensis.

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Section: Resultssupporting

confidence: 57%

IglG and IglI of the Francisella Pathogenicity Island Are Important Virulence Determinants of Francisella tularensis LVS

et al. 2011

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“…To date, only two F. tularensis OMPs have been described: FopA and TUL4 (22,27). This study describes, for the first time, the physical separation and enrichment of F. tularensis OMs from other cellular components for the identification of additional OMPs and LPs from both LVS and Schu S4.…”

Section: Discussionmentioning

confidence: 99%

“…The TUL4 paralogs share only 32% amino acid identity, with Tul4-B likely being the 17-kDa T-lymphocytereactive protein (41), based upon sequence homology (45) and predicted molecular mass (Table 1). Nevertheless, both proteins were selected for further analysis based upon previous reports of outer membrane localization of TUL4 (22).…”

Section: Bioinformatic Analysesmentioning

confidence: 99%

“…Initial [ 3 H]palmitate radiolabeling and detergent extraction of F. tularensis suggested that TUL4 was an integral membrane lipoprotein (LP) (46). Further studies later suggested that TUL4 most likely was in the F. tularensis OM (22). A number of other probable OMPs and LPs were identified in the aforementioned studies, yet none have been further characterized.…”

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confidence: 99%

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Characterization ofFrancisella tularensisOuter Membrane Proteins

et al. 2007

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Francisella tularensis is a gram-negative coccobacillus that is capable of causing severe, fatal disease in a number of mammalian species, including humans. Little is known about the proteins that are surface exposed on the outer membrane (OM) of F. tularensis, yet identification of such proteins is potentially fundamental to understanding the initial infection process, intracellular survival, virulence, immune evasion and, ultimately, vaccine development. To facilitate the identification of putative F. tularensis outer membrane proteins (OMPs), the genomes of both the type A strain (Schu S4) and type B strain (LVS) were subjected to six bioinformatic analyses for OMP signatures. Compilation of the bioinformatic predictions highlighted 16 putative OMPs, which were cloned and expressed for the generation of polyclonal antisera. Total membranes were extracted from both Schu S4 and LVS by spheroplasting and osmotic lysis, followed by sucrose density gradient centrifugation, which separated OMs from cytoplasmic (inner) membrane and other cellular compartments. Validation of OM separation and enrichment was confirmed by probing sucrose gradient fractions with antibodies to putative OMPs and inner membrane proteins. F. tularensis OMs typically migrated in sucrose gradients between densities of 1.17 and 1.20 g/ml, which differed from densities typically observed for other gram-negative bacteria (1.21 to 1.24 g/ml). Finally, the identities of immunogenic proteins were determined by separation on two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis and mass spectrometric analysis. This is the first report of a direct method for F. tularensis OM isolation that, in combination with computational predictions, offers a more comprehensive approach for the characterization of F. tularensis OMPs.

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“…В данной связи, ЛПС рассматривается как основной компонент молекулярной вакцины. В частности, согласно данным литературы, комплекс белков наружной мембраны и ЛПС, С-комплекс поверхностных структур F. tularensis обеспечивают определенный иммунитет лабораторных животных против возбудителя [2,15]. В то же время, в очищенном виде ни ЛПС, ни один из идентифицированных специфичных иммуногенных белков туляремийного микроба (например, Tul-4, Fop-A и др.)…”

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Role of Various Antigenic Preparations of Francisella Tularensis in Formation of Allergy Reaction in Humans and Animals

2016

Journal of microbiology, epidemiology and immunobiology

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Aim. Study the role of LPS in induction of anti-tularemia immunity in humans and animals. Materials and methods. Activity of various antigenic preparations of tularemia microbe, including highly purified from protein and S- and R-LPS, was studied using leukocytolysis reaction with blood of vaccinated humans and guinea pigs and skin allergy test (guinea pigs). Results. Only the whole cells of Francisella tularensis, killed in protein non-denaturating conditions and conserving full S-LPS structure (tularin+) were shown to be inductors of delayed-type hypersensitivity reaction. Alterations in LPS structure (tularin) results in a significant decrease, and denaturation of bacterial proteins (during boiling) results in a complete loss of immune stimulating properties of the preparations. Purified LPS preparations and O-polysaccharide fraction of S-LPS are not able to activate cell-mediated immunity. Conclusion. The presence of LPS with the full structure affects the ability of antigenic preparations of F.tularensis to cause allergic reactions, and thus, form cell-mediated anti-tularemia immunity. LPS of F.tularensis can not be excluded as an adjuvant and provides the most effective presentation of epitopes of protein molecules for interaction with receptors of T-lymphocytes.

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Outer membranes of a lipopolysaccharide-protein complex (LPS-17 kDa protein) as chemical tularemia vaccines

Cited by 5 publications

References 14 publications

IglG and IglI of the Francisella Pathogenicity Island Are Important Virulence Determinants of Francisella tularensis LVS

IglG and IglI of the Francisella Pathogenicity Island Are Important Virulence Determinants of Francisella tularensis LVS

Characterization ofFrancisella tularensisOuter Membrane Proteins

Role of Various Antigenic Preparations of Francisella Tularensis in Formation of Allergy Reaction in Humans and Animals

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