69 research outputs found

    Multifunctional profile of CD4<sup>+</sup> T cells in the lung of infant and adult mice following BCG immunization.

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    <p>Infant and adult mice were immunized s.c. with BCG and sacrificed at 4 (A), 8 (B), or 16 (C) weeks following immunization. Cells from the lung were stimulated with <i>M</i>.<i>tb</i> CF + crude BCG for 24h or left unstimulated as a control. Cells were stained and analyzed by flow cytometry. Average proportions displayed in pie chart are of the CD4 T cells expressing specific cytokine combinations. Absolute numbers of CD4<sup>+</sup> T cells in the tissues were calculated and displayed in bar graphs. Results are from one independent experiment per timepoint, n = 4-5/group/timepoint. Data are expressed as Mean ± SEM. *, p < 0.05; **, p < 0.005; ***, p < 0.0005.</p

    Consecutive Spray Drying to Produce Coated Dry Powder Vaccines Suitable for Oral Administration

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    Current global vaccination programs are challenged by costs associated with vaccine cold chain storage and administration. A solid, thermally stable oral dosage form for vaccines would alleviate these costs but is difficult to produce due to general vaccine instability and the complication of bypassing the gastric barrier. We developed a novel consecutive spray drying method that is suitable for use with biologics and employs Eudragit L100 polymer as the enteric coating. More specifically, in step 1, recombinant replication deficient human type-5 adenovirus and vesicular stomatitis virus were encapsulated by spray drying with sugars from a water solution, and in step 2, the microparticles from step 1 were suspended in ethanol with Eudragit and spray dried again. Up to 25% of the starting material was fully encapsulated within the enteric coating, and encapsulation efficiency was largely dependent on spray gas flow rate and the solids concentration in the feed. After step 2, the coated vaccine–sugar particles maintained their thermostability and were slightly larger in size with a rugous surface morphology compared to the particles produced in step 1. The coated particles retained viral vector activity in vitro after 15 min incubation in 1 M HCl (simulating the stomach environment) and anhydrous ethanol (to dissolve the Eudragit outer shell). The production of dry, orally administered vaccine particles from consecutive spray drying offers the potential to remedy a number of vaccine storage, transportation, and administration limitations

    Functional networks most significanty modulated in PPD-B stimulated PBMC from vaccinated/protected calves.

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    <p>Visualisation of the trend and significance of each network: Red bars = up-regulation; blue bars = down-regulation of network. Horizontal bar: p = 0.05.</p

    Ag-specific T cell responses in the lung after boost AdHu5Ag85A immunization in BCG primed mice.

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    <p>Infant and adult mice were BCG immunized, and the AdHu5Ag85A booster vaccine was administered i.n. at 16 weeks post-BCG (A). The mice were sacrificed 4 weeks (B, C, D) or 8 weeks (E, F, G) after boosting. Lung cells were stimulated either with <i>M</i>.<i>tb</i> CF + crude BCG for 24h (open bar), or Ag85A-specific CD4 or CD8 T cell peptide for 6h (black bar), or left unstimulated (B, C, E, F). Cells were stained and analyzed by flow cytometry. Absolute numbers of IFN-γ<sup>+</sup>CD4<sup>+</sup> (B, E) and IFN-γ<sup>+</sup>CD8<sup>+</sup> (C, F) T cells were calculated (unstimulated subtracted from stimulated). Ag85A CD8 peptide tetramer staining was performed on lung cells, and analyzed by flow cytometry (D, G). Absolute numbers of tet<sup>+</sup>CD8<sup>+</sup> T cells were calculated. Results are from one experiment per timepoint, n = 4-5/group/timepoint. Data are expressed as Mean ± SEM. *, p < 0.05; **, p < 0.005; ***, p < 0.0005. All other comparisons (not indicated) were not significant.</p

    CD11c APC populations were isolated from BAL, lung parenchyma and the spleen and pulsed with M

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    Tb CF protein or infected with live mycobacteria. APCs were then co-cultured with CD4+ or CD8+ T cells that were purified from the mice that were infected by live mycobacteria for 17 days (diagram). Cells were co-cultured for 24 h and IFN-γ-secreting T cells were determined by ELISPOT assay. A, IFN-γ-secreting CD4+ T cells. B, IFN-γ-secreting CD8+ T cells. Data are expressed as the mean value ± SD of triplicate samples and representative of two independent experiments. ‡p < 0.05 compared to the corresponding mycobacterial BCG-infected APCs; #p < 0.05 compared to the corresponding spleen data.<p><b>Copyright information:</b></p><p>Taken from "CD11c+ antigen presenting cells from the alveolar space, lung parenchyma and spleen differ in their phenotype and capabilities to activate naïve and antigen-primed T cells"</p><p>http://www.biomedcentral.com/1471-2172/9/48</p><p>BMC Immunology 2008;9():48-48.</p><p>Published online 13 Aug 2008</p><p>PMCID:PMC2527294.</p><p></p

    CD11c APC populations isolated from BAL, lung and spleen were infected with AdOVA and co-cultured either for 48 hours with CFSE-labelled OT-I CD8 T cells or for 72 hours with CFSE-labelled OT-II CD4 T cells

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    The rate of transgenic T cell proliferation was analyzed by FACS. A, OT-I CD8 T cell proliferation. B, OT-II CD4 T cell proliferation. Representative histograms of T cell proliferation are shown. The average T cell proliferation rates are shown in bar graphs after subtracting from appropriate controls. Results are presented as the mean ± SD of triplicate samples. *p < 0.05.<p><b>Copyright information:</b></p><p>Taken from "CD11c+ antigen presenting cells from the alveolar space, lung parenchyma and spleen differ in their phenotype and capabilities to activate naïve and antigen-primed T cells"</p><p>http://www.biomedcentral.com/1471-2172/9/48</p><p>BMC Immunology 2008;9():48-48.</p><p>Published online 13 Aug 2008</p><p>PMCID:PMC2527294.</p><p></p

    Freshly isolated CD11c APCs from the BAL, lung parenchyma, and the spleen were cultured for 48 h with LPS or mycobacterial antigens (Mtb-CF)

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    Culture supernatants were measured for TNF-α (A) and IL-12 (B) by ELISA. Results are representative of three independent experiments and are presented as mean ± SD. *p < 0.05 compared to the corresponding lung data. #p < 0.05 compared to the corresponding spleen data.<p><b>Copyright information:</b></p><p>Taken from "CD11c+ antigen presenting cells from the alveolar space, lung parenchyma and spleen differ in their phenotype and capabilities to activate naïve and antigen-primed T cells"</p><p>http://www.biomedcentral.com/1471-2172/9/48</p><p>BMC Immunology 2008;9():48-48.</p><p>Published online 13 Aug 2008</p><p>PMCID:PMC2527294.</p><p></p

    Results of RNA-Seq analysis.

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    <p>A. Signficantly modulated genes in the three treatment groups. B. Venn diagrams of genes significantly up-regulated and (C) down-regulated genes after vaccination but prior to <i>M. bovis</i> challenge. A. Fold change compared to unstimulated PBMC (medium controls) of PPD-B stimulated PBMC compared to medium controls from unvaccinated, naïve calves (group 1), vaccinated/non-protected (group 2), and vaccinated/protected calves (group 3).</p

    Effect of elapsed time between BCG priming and AdHu5Ag85A boost immunization on Ag-specific responses boosted by AdHu5Ag85A boost immunization.

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    <p>Infant and adult mice were BCG immunized, and boosted with AdHu5Ag85A at 8 or 16 weeks post-BCG (A). The mice were sacrificed 4 weeks after boosting. Cells isolated from the lung were stimulated either with <i>M</i>.<i>tb</i> CF + crude BCG (B), Ag85A-specific CD4 T cell peptide (C), or CD8 T cell peptide (D), or left unstimulated as a control. Cells were stained and analyzed by flow cytometry. Absolute numbers of IFN-γ<sup>+</sup>CD4<sup>+</sup> (B, C) and IFN-γ<sup>+</sup>CD8<sup>+</sup> (D) T cells were calculated (unstimulated subtracted from stimulated). Ag85A CD8 peptide tetramer staining was performed on lung cells, and analyzed by flow cytometry (E). Absolute numbers of tet<sup>+</sup>CD8<sup>+</sup> T cells were calculated. Results are from one experiment per timepoint, n = 4-5/group/timepoint. Data are expressed as Mean ± SEM. *, p < 0.05; **, p < 0.005; ***, p < 0.0005. All other comparisons (not indicated) were not significant.</p
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