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

<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⁺ 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

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

<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.

<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⁺CD4⁺ (B, E) and IFN-γ⁺CD8⁺ (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⁺CD8⁺ 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 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

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

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

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

Results of RNA-Seq analysis.

<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.

<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⁺CD4⁺ (B, C) and IFN-γ⁺CD8⁺ (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⁺CD8⁺ 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

Experimental regimens used for both short-term and long-term guinea pig studies.

<p>Experimental regimens used for both short-term and long-term guinea pig studies.</p

Total # of IFN-γ⁺ T cells in BAL in mice boosted at 16 weeks post-BCG (x 10³).

<p>Infant and adult mice were immunized s.c. with BCG, and the AdHu5Ag85A booster vaccine was administered intranasally (i.n.) at 16 weeks post-BCG. The mice were sacrificed 4 or 8 weeks after boosting. Cells isolated from the BAL were stimulated with <i>M</i>.<i>tb</i> CF + crude BCG, or unstimulated as a control. Cells were stained with extracellular antibodies for T cell markers, followed by intracellular staining for IFN-γ, and analyzed by flow cytometry. Absolute numbers of IFN-γ⁺CD4⁺ and IFN-γ⁺CD8⁺ T cells were calculated (unstimulated subtracted from stimulated). Results are from one experiment per timepoint, n = 4-5/group/timepoint. Data are expressed as Mean ± SEM.</p><p>*, p < 0.05.</p><p>Total # of IFN-γ⁺ T cells in BAL in mice boosted at 16 weeks post-BCG (x 10³).</p