Proteomic analysis of anti-Francisella tularensis LVS antibody response in murine model of tularemia

Havlasová, J; Hernychová, Lenka; Brychta, Martin; Hubálek, Martin; Lenčo, Juraj; Larsson, Pär; Lundqvist, Margaretha; Forsman, Mats; Krocova, Z; Stulı́k, Jiřı́; Macela, Aleš

doi:10.1002/pmic.200401123

Cited by 72 publications

(74 citation statements)

References 56 publications

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“…The latter study identified a larger number of potential OMP candidates, approximately 500 in total, yet only 4 were predicted to be OMPs, including FopA. In an effort to identify immunogenic proteins from F. tularensis, total cellular proteins were separated by 2DE and probed with sera from experimentally infected mice (19). Of the 36 immunoreactive proteins identified, the vast majority were probable cytoplasmic proteins, whereas only two predicted OMPs were noted.…”

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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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Characterization ofFrancisella tularensisOuter Membrane Proteins

et al. 2007

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“…Cross-reactions are possible only with serum obtained from patients with brucellosis or yersiniosis, but sera with a titer lower than 1:320 do not agglutinate Brucellae at all [129,197]. The agglutination test can be followed by the Rose Bengal plate test, which is often used as a rapid screening test in the diagnosis of brucellosis [198], or, according to our experience, with immunoproteomic techniques to exclude cross-reactions [194].…”

Section: Diagnosis Detection and Laboratory Confirmationmentioning

confidence: 99%

“…Among these are antibodies oriented against some outer membrane proteins such as FopA, OmpA [192,193], and Tul4 [194]; against other intracellular proteins such as GroEL, KatG [192], and DnaK; and against several putative virulence markers such as nucleoside diphosphate kinase, isocitrate dehydrogenase, the RNA-binding protein Hfq, and the molecular chaperone ClpB [194,195]. Thus, antibodies can potentially contribute to the protective response by eliminating Francisella virulence factors and, together with the antibody-independent functions of B cells, can demonstrate the potential of B cells to collaborate with T cells in the induction, regulation, and expression of protective immunity against F. tularensis infection.…”

Section: Host Immune Responsementioning

confidence: 99%

Putting the Jigsaw Together - A Brief Insight Into the Tularemia

Kubelková

Macela

2015

Open Life Sciences

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Tularemia is a debilitating febrile and potentially fatal zoonotic disease of humans and other vertebrates caused by the Gram-negative bacterium Francisella tularensis. The natural reservoirs are small rodents, hares, and possibly amoebas in water. The etiological agent, Francisella tularensis, is a non-spore forming, encapsulated, facultative intracellular bacterium, a member of the γ-Proteobacteria class of Gram-negative bacteria. Francisella tularensis is capable of invading and replicating within phagocytic as well as non-phagocytic cells and modulate inflammatory response. Infection by the pulmonary, dermal, or oral routes, respectively, results in pneumonic, ulceroglandular, or oropharyngeal tularemia. The highest mortality rates are associated with the pneumonic form of this disease. All members of Francisella tularensis species cause more or less severe disease Due to their abilities to be transmitted to humans via multiple routes and to be disseminated via biological aerosol that can cause the disease after inhalation of even an extremely low infectious dose, Francisella tularensis has been classified as a Category A bioterrorism agent. The current standard of care for tularemia is treatment with antibiotics, as this therapy is highly effective if used soon after infection, although it is not, however, absolutely effective in all cases.

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“…Finally, ABAYE3267 protein sequence is similar to a highly conserved protein, the nucleoside diphosphate kinase (NDK) that is present in a number of bacteria including Bacillus anthracis, Francisella tularensis and Campylobacter fetus and considered to be a potential virulence target and thus a vaccine candidate. [54][55][56] Using the MBGD platform, 34 the proteomic data of OMVs obtained from HPLC/MS/MS experiments and bioinformatics prediction of surface-exposed antigens, we have identified 77 antigens as potential vaccine candidates ( Table S2). The final list was restricted to 13 putative antigens.…”

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confidence: 99%