8 research outputs found

    The <i>emrA1</i> mutant vaccinated mice induce sustained production of pro-inflammatory cytokines and a potent antibody response following lethal <i>Ft</i> LVS challenge.

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    <p>C57BL/6 mice immunized i.n. with 1×10<sup>6</sup> CFU of the <i>emrA1</i> mutant were challenged i.n. with 1×10<sup>7</sup> CFU of wild type <i>Ft</i> LVS 42 days post-immunization. <b>(A-D)</b> On days 5, 7 and 14 post-challenge, mice (n = 3 per group/time point) were euthanized and their excised lungs were homogenized. Clear lung homogenates were used for quantification of indicated pro-inflammatory cytokines using flow cytometric analysis. The data are represented as Mean ± S.D. <b>(E)</b> On day 21 post-challenge, mice (n = 3 per group) were anesthetized and bled retroorbitally to obtain serum. <i>Ft</i> specific total IgG, IgG2a, IgG2b, IgG1 and IgA levels in serum samples were determined by ELISA. The data are represented as Mean ± S.D. of absorbance values measured at 450 nm. Red arrows indicate antibody titers. ND = Not detected.</p

    Mice immunized with the <i>emrA1</i> mutant are protected against 1000LD100–10,000LD100 challenge dose of <i>Ft</i> LVS.

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    <p>C57BL/6 mice (n = 5–10 per group) were immunized i.n. with 1×10<sup>6</sup> CFU of the <i>emrA1</i> mutant. <b>(A, B)</b> On day 42 of the primary immunization mice were challenged i.n. with 1×10<sup>7</sup> CFU of wild type <i>Ft</i> LVS. Age matched unvaccinated mice challenged with a similar dose of <i>Ft</i> LVS were kept as controls. (<b>C, D)</b> On day 42 of the primary immunization mice were challenged i.n. with 1×10<sup>8</sup> CFU of wild type <i>Ft</i> LVS. Age matched unvaccinated mice challenged with a similar dose of <i>Ft</i> LVS were kept as controls. (<b>E, F)</b> On day 75 of the primary immunization mice were challenged i.n. with 1×10<sup>7</sup> CFU of wild type <i>Ft</i> LVS. Age matched unvaccinated mice challenged with a similar dose of <i>Ft</i> LVS were kept as controls. The Challenged mice were observed for morbidity and mortality for a period of 21 days post-challenge (<b>A, C, E</b>). The mice were weighed at the indicated times post-challenge to monitor the progression of infection (<b>B, D, F</b>). The survival results are expressed as Kaplan-Meier survival curves and P values were determined by Log-rank test. Body weights of mice are expressed percent body weights.</p

    Immunization with the <i>emrA1</i> mutant results in minimal weight loss, rapid bacterial clearance, and histopathological lesions in lung, liver and spleen.

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    <p>C57BL/6 mice were immunized i.n. with 1×10<sup>6</sup> CFU of the <i>emrA1</i> mutant. Mice infected with equal numbers of wild type <i>Ft</i> LVS were used as controls. <b>(A)</b> The immunized mice were weighed at the indicated times post-immunization to track the progress of infection. <b>(B)</b> On days 1, 5, 7, 14 and 21 post-immunization, mice (n = 4 per group/time point) were euthanized and bacterial burdens were quantified in their lung, liver and spleen. Bacterial counts in organs are expressed as Log<sub>10</sub>CFU/mL. The <i>P</i> values were determined using one way ANOVA. *<i>P<0</i>.<i>05; **P<0</i>.<i>01; ***P<0</i>.<i>001</i>. <b>(C)</b> Excised lungs, livers and spleens were preserved in 10% formalin, paraffin embedded, sliced into 5 μM thin sections and stained with Hematoxylene & Eosin. Stained sections were observed for histopathological lesions under a light microscope (Magnification 100×). # = <i>Ft</i> LVS infected mice succumbed to infection.</p

    Immunization with <i>emrA1</i> mutant using a prime-boost vaccination regimen improves the extent of protection against <i>Ft</i> SchuS4 challenge.

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    <p><b>(A)</b> C57BL/6 mice (n = 10 per group) were immunized i.n. with 1×10<sup>6</sup> CFU of the <i>emrA1</i> mutant and boosted i.d. on day 21 with a similar dose. On day 42 of the primary immunization mice were challenged i.n. with 17 CFU of <i>Ft</i> SchuS4. Age matched unvaccinated mice challenged with a similar dose of <i>Ft</i> SchuS4 served as controls. <b>(B)</b> C57BL/6 mice (n = 10 per group) were immunized i.d. with 1×10<sup>6</sup> CFU of the <i>emrA1</i> mutant and boosted i.n. on day 21 with a similar dose. On day 42 of the primary immunization mice were challenged i.n. with 17 CFU of <i>Ft</i> SchuS4. Age matched unvaccinated mice challenged with a similar dose of <i>Ft</i> SchuS4 served as controls. The survival results are expressed as Kaplan-Meier survival curves and P values were determined by Log-rank test.</p

    Mice immunized with the <i>emrA1</i> mutant induce regulated production of pro-inflammatory cytokines and a potent antibody response.

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    <p>C57BL/6 mice were immunized i.n. with <i>1</i>×10<sup>6</sup> CFU of the <i>emrA1</i> mutant or <i>Ft</i> LVS. On days 1, 5, 7 and 14 post-immunization, mice (n = 4 per group/time point) were euthanized and their excised lungs and spleens were homogenized. Clear lung <b>(A-D)</b> and spleen <b>(E-H)</b> homogenates were used for quantification of indicated pro-inflammatory cytokines using flow cytometric analysis. The data are represented as Mean ± S.D. The <i>P</i> values were determined using one way ANOVA. *<i>P<0</i>.<i>05; **P<0</i>.<i>01</i>. <b>(I)</b> On day 42 post-immunization, mice (n = 3 per group/ time point) were anesthetized and bled retroorbitally to obtain serum. <i>Ft</i> specific total IgG, IgG2a, IgG2b, IgG1 and IgA levels in serum samples were determined by ELISA. The data are represented as Mean ± S.D. of absorbance values measured at 450 nm. Red arrows indicate antibody titers. # = <i>Ft</i> LVS immunized mice succumbed to infection; ND = Not detected.</p

    Immunization with <i>emrA1</i>-mAb complexes improves the extent of protection against <i>Ft</i> SchuS4 challenge.

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    <p><b>(A)</b> C57BL/6 mice (n = 10 per group) immunized i.n. with 1×10<sup>6</sup> CFU of the <i>emrA1</i> mutant-mAb immune complexes. On day 42 of the primary immunization mice were challenged i.n. with 32 CFU of <i>Ft</i> SchuS4. Age matched unvaccinated mice challenged with a similar dose of <i>Ft</i> SchuS4 served as controls. The survival results are expressed as Kaplan-Meier survival curves and P values were determined by Log-rank test. <b>(B)</b> The mice were weighed at the indicated times post-challenge to monitor the progression of infection. <b>(C)</b> The indicated <i>Ft</i> specific antibodies were determined in serum from immunized mice on day 14 post-immunization. The results are expressed as antibody titers.</p

    The <i>emrA1</i> mutant vaccinated mice clear bacteria rapidly and exhibit minimal histopathological lesions in lung, liver and spleen following lethal <i>Ft</i> LVS challenge.

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    <p>C57BL/6 mice immunized i.n. with 1×10<sup>6</sup> CFU of the <i>emrA1</i> mutant or the unvaccinated control mice were challenged i.n. with 1×10<sup>7</sup> CFU of <i>Ft</i> LVS 42 days post-immunization. <b>(A)</b> On days 5, 7, and 14 post-challenge, mice (n = 3 per group/time point) were euthanized and bacterial burdens were quantified in their lung, liver and spleen. Bacterial counts in organs are expressed as Log<sub>10</sub> CFU/mL. The <i>P</i> values were determined using one way ANOVA. **<i>P<0</i>.<i>01;</i> ***<i>P<0</i>.<i>001</i>. <b>(B)</b> Lungs, livers and spleens collected at the indicated times post-challenge were preserved in 10% formalin, embedded in paraffin blocks, sliced into 5 μM thin sections and stained with Hematoxylene & Eosin. Stained sections were observed for histopathological lesions under a light microscope (Magnification 100×). # = Unvaccinated mice succumbed to infection.</p

    Immunization with <i>emrA1</i>-mAb complexes using a prime-boost vaccination regimen or a low dose immunization improves the extent of protection against <i>Ft</i> SchuS4 challenge.

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    <p><b>(A)</b> C57BL/6 mice (n = 10 per group) were immunized i.d. with 1×10<sup>6</sup> CFU of the <i>emrA1</i> mutant-mAb immune complex and boosted i.n. on day 21 with a similar dose. On day 42 of the primary immunization mice were challenged i.n. with 17 CFU of <i>Ft</i> SchuS4. Age matched unvaccinated mice challenged with a similar dose of <i>Ft</i> SchuS4 served as controls. <b>(B)</b> C57BL/6 mice (n = 10 per group) immunized i.n. with 1×10<sup>6</sup> CFU of the <i>emrA1</i>mutant-mAb immune complex and boosted i.d. on day 21 with a similar dose. On day 42 of the primary immunization mice were challenged i.n. with 17 CFU of <i>Ft</i> SchuS4. Age matched unvaccinated mice challenged with a similar dose of <i>Ft</i> SchuS4 served as controls. <b>(C)</b> C57BL/6 mice (n = 10 per group) immunized i.n. with 1×10<sup>3</sup> CFU of the <i>emrA1</i> mutant and boosted i.d. on day 21 with a similar dose. On day 42 of the primary immunization mice were challenged i.n. with 17 CFU of <i>Ft</i> SchuS4. Age matched unvaccinated mice challenged with a similar dose of <i>Ft</i> SchuS4 served as controls. <b>(D)</b> C57BL/6 mice (n = 10 per group) immunized i.d. with 1×10<sup>3</sup> CFU of the <i>emrA1</i> mutant and boosted i.n. on day 21 with a similar dose. On day 42 of the primary immunization mice were challenged i.n. with 17 CFU of <i>Ft</i> SchuS4. Age matched unvaccinated mice challenged with a similar dose of <i>Ft</i> SchuS4 served as controls. The survival results are expressed as Kaplan-Meier survival curves and P values were determined by Log-rank test.</p
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