Levels of cytokines secreted by human PBMC after recall stimulation with ffLVS or ffSchu S4 for five days.

<p>Cytokine concentrations were measured in cell culture supernatants using multiplex analysis. Median values ± SEM from PBMC samples of 14–16 individuals per donor group are shown (black bars indicate convalescent patients; grey bars indicate LVS vaccinees; white bars indicate naïve donors). Statistically significant differences between immune and naïve donors are marked by asterisks (<i>P</i><0.05). For IL-7, 36 out of 40 values were below the detection limit and therefore not included in the data analysis.</p

Histograms indicating the frequency of mono-, bi- and tri-functional cell subsets in CD45RO⁺ and CD45RA⁺ T-cell populations of convalescent patients.

<p>Mean values are illustrated throughout. The black axis indicates percentages for mono-functional IFN-γ-positive and all bi- and tri-functional T-cell subsets. The red axis indicates percentages for MIP-1β- and CD107a-positive mono-functional T cell subsets.</p

Stimulation indices for the proliferative responses of human PBMC to recall stimulation with ffLVS for five days.

<p>Median values ± SEM per donor group of 12–16 individuals are shown. “pat” indicates convalescent tularemia patients, “vc” LVS vaccinees, and “nv” naïve individuals.</p

Heat map of Spearman correlation coefficients for iMFI values in various cell populations after intra-individual comparison for all donors.

<p>Integrated MFI values were obtained for all three functional markers (IFN-γ, MIP-1β, CD107a) in all mono-, bi- and trifunctional T-cell subsets and for all donors using <i>Clust</i> semi-automated gating. The iMFI values (24 values per PBMC sample, identified by numbers 1–24) from recall stimulation with 0.1 cfu ffLVS/PBMC were compared and lined up in a two-dimensional matrix. Correlated data are marked by shades of grey depending on the strength of the correlation coefficients; a coefficient above 0.7 is considered to indicate very strong correlation.</p

Histograms indicating the frequency of mono-, bi- and tri-functional cell subsets in CD45RO⁺ and CD45RA⁺ T-cell populations of vaccinees.

<p>Mean values are illustrated throughout. The black axis indicates percentages for mono-functional IFN-γ-positive and all bi- and tri-functional T-cell subsets. The red axis indicates percentages for MIP-1β- and CD107a-positive mono-functional T cell subsets.</p

Histograms indicating the frequency of mono-, bi- and tri-functional cell subsets in CD45RO⁺ and CD45RA⁺ T-cell populations of non-vaccinated individuals.

<p>Mean values are illustrated throughout. The black axis indicates percentages for mono-functional IFN-γ-positive and all bi- and tri-functional T-cell subsets. The red axis indicates percentages for MIP-1β- and CD107a-positive mono-functional T cell subsets.</p

Levels of cytokines secreted by human PBMC after recall stimulation with ffLVS or ffSchu S4 for five days.

<p>Cytokine concentrations were measured in cell culture supernatants using multiplex analysis. Median values ± SEM from PBMC samples of 14–16 individuals per donor group are shown (black bars indicate convalescent patients; grey bars indicate LVS vaccinees; white bars indicate naïve donors). Statistically significant differences between immune and naïve donors are marked by asterisks (<i>P</i><0.05).</p

Immunoblot analysis of the filtrates for the detection of a cytoplasmic marker, cAMP receptor protein

<p><b>Copyright information:</b></p><p>Taken from "Vesicle-independent extracellular release of a proinflammatory outer membrane lipoprotein in free-soluble form"</p><p>http://www.biomedcentral.com/1471-2180/8/18</p><p>BMC Microbiology 2008;8():18-18.</p><p>Published online 28 Jan 2008</p><p>PMCID:PMC2257964.</p><p></p> The whole cell protein preparations of strain (lane 1), strain (lane 2) and D7SS (lane 3) were used as controls in the immunoblot with antibodies against cAMP receptor protein. Serum filtrates (8 h): D7SS (lane 4), D7SS-p (lane 5) and No bacteria control (lane 6). Broth filtrates (24 h): D7SS (lane 7), D7SS-p (lane 8) and No bacteria control (lane 9). Detection of cAMP receptor protein from precipitated filtrates of lysed (lane 10) and unlysed D7SS cells (lane 11) incubated in the inserts containing broth for 8 h

Immunoblot detection of PAL release from cells exposed to heat-inactivated FCS or cultured in broth

<p><b>Copyright information:</b></p><p>Taken from "Vesicle-independent extracellular release of a proinflammatory outer membrane lipoprotein in free-soluble form"</p><p>http://www.biomedcentral.com/1471-2180/8/18</p><p>BMC Microbiology 2008;8():18-18.</p><p>Published online 28 Jan 2008</p><p>PMCID:PMC2257964.</p><p></p> The samples were obtained from cell-culture wells outside the inserts, where the bacteria [D7SS (lane 1) and D7SS-p (lane 2)] or No bacteria (lane 3) were incubated in serum for 2 h and 8 h (Panel A) or cultured in broth for 24 h (Panel B). AaPAL was detected by immunoblot using anti-AaPAL peptide antiserum. Samples for bacterial culture and enumeration were taken from the inserts containing serum or broth (Panel C) at the same time points as the samples for the immunoblots in Panels A and B. The results show means (SD) from three independent experiments