40 research outputs found

    Immunization with the <i>FTL_0325</i> mutant induces an early pro-inflammatory cytokine response in lungs.

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    <p>BALB/c mice (n = 4) were infected with 1×10<sup>7</sup> CFU of the indicated mutants and wild type <i>F. tularensis</i> LVS. At the indicated times, the levels of pro-inflammatory cytokines were measured in lung homogenates using a Cytometric Bead Array assay. The data are representative of two independent experiments conducted and were analyzed using ANOVA with Tukey-Kramer Multiple Comparison post-test and <i>P</i> values were recorded. <i>**P</i><0.01; ***<i>P<</i>0.001. Ψ = Mice infected with 1×10<sup>7</sup> CFU of <i>F. tularensis</i> LVS succumbed to infection by day 7 PI and hence were unavailable for comparison.</p

    <i>FTL_0304</i>, <i>FTL_0291</i>, <i>FTL_0325</i> and <i>FTL_0057</i> mutants are highly attenuated for virulence in BALB/c and C57BL/6 mice.

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    a<p>6–8 weeks old mice were infected i.n. with either <i>F. tularensis</i> LVS or the indicated mutants and monitored for mortality for 28 days. Data are shown as number of mice survived/total number of mice infected.</p

    Immunization with the <i>FTL_0325</i> mutant results in a potent memory recall response.

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    <p>Cell culture supernatants collected from BMDMs infected with either the <i>F. tularensis</i> LVS (A–D) or <i>F. tularensis</i> SchuS4 (E and F) and co-cultured with the splenocytes prepared from immunized mice at day 45 post-immunization (A and C) or at day 60 (B–F) were analyzed for IFN-γ (A, B and E) and IL-17a (C,D and F) cytokines by Cytometric Bead Flex sets. The data are representative of two independent experiments and were analyzed using ANOVA with Tukey-Kramer Multiple Comparison post-test and <i>P</i> values recorded. *<i>P<</i>0.05<i>; **P</i><0.01; ***<i>P<</i>0.001.</p

    Immunization with the <i>FTL_0325</i> mutant induces a higher pro-inflammatory cytokine response in the spleen.

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    <p>BALB/c mice (n = 4) were infected with 1×10<sup>7</sup> CFU of the indicated mutants and wild type <i>F. tularensis</i> LVS. At the indicated times, the levels of pro-inflammatory cytokines were measured in spleen homogenates using a Cytometric Bead Array assay. The data are representative of two independent experiments conducted and were analyzed using ANOVA with Tukey-Kramer Multiple Comparison post-test and <i>P</i> values were recorded. *<i>P<</i>0.05<i>; **P</i><0.01; ***<i>P<</i>0.001. Ψ = Mice infected with 1×10<sup>7</sup> CFU of <i>F. tularensis</i> LVS succumbed to infection by day 7 PI and hence were unavailable for comparison.</p

    Priming enhances BMDMs responses to non-HAd, but not HAd, <i>Ft</i>.

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    <p>(A) Primary BMDMs and (B) immortalized BMDMs (IMC) from wild-type and TLR2<sup>−/−</sup> C57BL/6 mice (2.5×10<sup>5</sup> cells/well) were treated with IFN-γ (50 ng/well) for 16 h prior to infection with MHB- and BHI-grown <i>Ft</i> LVS at a MOI of 100. (C) In a separate set of experiments priming was achieved by incubation of cells with 4% autologous serum for 16 h prior to infection with <i>Ft</i> LVS. The autologous serum was recovered from <i>Ft</i> LVS-infected mice at day 6 p.i. For both sets of experiments, supernatants were collected 24 h p.i. and were assayed for the presence of cytokines by CBA. Results represent the mean ± SEM from three independent experiments. **<i>P</i><0.01, ***<i>P</i><0.001. (All results shown were subjected to One-way ANOVA with Bonferroni's Post-test).</p

    HAd-<i>Ft</i> LVS is incapable of eliciting T<sub>H</sub>1-type pro-inflammatory responses irrespective of the cell-type.

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    <p>BMDMs (A & B), BMD-dendritic cells (BMD-DCs) (C), alveolar macrophage MHS cell line (D), and human peripheral blood-derived macrophages (E) were infected at a MOI of 100 with <i>Ft</i> grown in MHB or BHIB or bacteria derived from macrophages. Regardless of cell-type, 2.5×10<sup>5</sup> cells were plated per well, supernatants were collected after 24 h and analyzed for the presence of cytokines by CBA. Results represent the mean ± SEM from two independent experiments. *<i>P</i><0.05, ***<i>P</i><0.001. (One-way ANOVA with Bonferroni's Post-test).</p

    Immunization with the <i>FTL_0325</i> mutant induces an early inflammatory response.

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    <p>BALB/c mice were infected with 1×10<sup>7</sup> CFU of the <i>F. tularensis</i> LVS, <i>FTL_0325</i> or <i>FTL_0304</i> mutant and the lungs were harvested at the indicated times, sectioned and stained with H & E. Representatives of H & E stained lung sections are shown. Infiltration of neutrophils is shown in the inset (Magnification 10×; Inset 100×). Ψ = Mice infected with 1×10<sup>7</sup> CFU of <i>F. tularensis</i> LVS succumbed to infection by day 7 PI and hence were unavailable for comparison. Green arrows indicate the sites of cellular infiltration. Yellow circles in inset show neutrophilic infiltration in the lungs of <i>FTL_0325</i> mutant immunized mice.</p

    The <i>FTL_0325</i> and <i>FTL_0304</i> mutants of <i>F. tularensis</i> LVS exhibit no growth defect under acellular growth conditions.

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    <p>Growth curves for bacteria grown in MHB were generated in a 96-well plate using 200 µl culture volumes while the growth curves for bacteria grown in BHI and CDM were generated using a 25 ml culture volume. Results shown are representative of two independent experiments.</p

    OmpA, DnaK and Tul4 Specific Antibody Responses in Mice Immunized with TMV-Multiconjugate Vaccines using Schedule I and II of Immunization.

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    <p><i>F</i>. <i>tularensis</i> SchuS4 recombinant proteins OmpA, DnaK and Tul4 specific IgG, antibody levels on day 28 in serum samples of C57BL/6 mice immunized with TMV-multiconjugate vaccine using Schedule II were determined by ELISA. The plates were coated with recombinant <i>F</i>. <i>tularensis</i> SchuS4 proteins. Serum samples obtained from naïve mice or those inoculated with TMV alone were used as controls. The data are represented as Mean ±S.D. of absorbance values measured at 450nm. The comparisons are shown with the data obtained from mice immunized with TMV-multiconjugate vaccine using schedule I (shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0130858#pone.0130858.g007" target="_blank">Fig 7</a>). Table shows comparison of antibody titers between groups of mice vaccinated with Schedule I and II vaccination regimens.</p
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