Dendritic Cells are preferentially targeted among hematolymphocytes by Modified Vaccinia Virus Ankara and play a key role in the induction of virus-specific T cell responses -5

S for 4 h at either 37 C° or 4 C°. Phagocytosis of MVA-infected DCs by uninfected DCs were detected by flow cytometry. This data is representative of two independent experiments.<p><b>Copyright information:</b></p><p>Taken from "Dendritic Cells are preferentially targeted among hematolymphocytes by Modified Vaccinia Virus Ankara and play a key role in the induction of virus-specific T cell responses "</p><p>http://www.biomedcentral.com/1471-2172/9/15</p><p>BMC Immunology 2008;9():15-15.</p><p>Published online 15 Apr 2008</p><p>PMCID:PMC2359732.</p><p></p

Dendritic Cells are preferentially targeted among hematolymphocytes by Modified Vaccinia Virus Ankara and play a key role in the induction of virus-specific T cell responses -7

Peated on day 2 post infection. WT littermates were included as controls. At day 6 following the infection, spleens were harvested to prepare single cell suspension. (A) The efficiency of DC depletion was determined by surface cell lineage marker staining and flow cytometry. Numbers were the percentages of the gated population in total viable splenocytes. (B, C) Spleen single cell suspension (effector) was re-stimulated with MVA-infected C57BL/6 splenocytes (target) for 12 h in the presence of Brefeldin A. Following incubation, the frequencies of IFN-γ-producing T cells in the effector splenocytes were determined by intracellular cytokine staining, as described in Methods. Numbers were the percentages of IFN-γcells in CD3T cell population. (C) The average ± SD percentages of IFN-γcells for each group (n = 4). Data are representative of two independent experiments. **: P < 0.01.<p><b>Copyright information:</b></p><p>Taken from "Dendritic Cells are preferentially targeted among hematolymphocytes by Modified Vaccinia Virus Ankara and play a key role in the induction of virus-specific T cell responses "</p><p>http://www.biomedcentral.com/1471-2172/9/15</p><p>BMC Immunology 2008;9():15-15.</p><p>Published online 15 Apr 2008</p><p>PMCID:PMC2359732.</p><p></p

Dendritic Cells are preferentially targeted among hematolymphocytes by Modified Vaccinia Virus Ankara and play a key role in the induction of virus-specific T cell responses -8

Enocytes were used as negative control. (B) BALB/c mice were either uninfected or infected with rMVA-GFP at 3 × 10PFU/mouse by i.v. injection. Spleens were harvested at 9 h post infection and GFP expression in various subsets of splenocytes was monitored by flow cytometry. DCs: CD11c; macrophages: CD1bCD11c; B cells: CD19; CD8T cells: CD8CD11c; CD4T cells: CD4CD11c. Numbers shown are the percentages (average ± standard deviation (SD)) of GFPcells in the corresponding cell subsets. The data are representative of three independent experiments.<p><b>Copyright information:</b></p><p>Taken from "Dendritic Cells are preferentially targeted among hematolymphocytes by Modified Vaccinia Virus Ankara and play a key role in the induction of virus-specific T cell responses "</p><p>http://www.biomedcentral.com/1471-2172/9/15</p><p>BMC Immunology 2008;9():15-15.</p><p>Published online 15 Apr 2008</p><p>PMCID:PMC2359732.</p><p></p

Dendritic Cells are preferentially targeted among hematolymphocytes by Modified Vaccinia Virus Ankara and play a key role in the induction of virus-specific T cell responses -4

Were harvested and viable cells were counted with trypan blue exclusion method. These data represent the average ± SD viability of 6 replicates from 2 experiments. (B) Immature BMDCs and LPS-stimulated mature BMDCs were mock treated or infected with rMVA-GFP at a MOI of 10. Twenty four hours later, cell viability was examined by trypan blue exclusion. These data represent the average (± SD) viability of 15 replicates from 5 experiments. NS: statistically non-significant; *: P < 0.05; **: P < 0.01.<p><b>Copyright information:</b></p><p>Taken from "Dendritic Cells are preferentially targeted among hematolymphocytes by Modified Vaccinia Virus Ankara and play a key role in the induction of virus-specific T cell responses "</p><p>http://www.biomedcentral.com/1471-2172/9/15</p><p>BMC Immunology 2008;9():15-15.</p><p>Published online 15 Apr 2008</p><p>PMCID:PMC2359732.</p><p></p

Dendritic Cells are preferentially targeted among hematolymphocytes by Modified Vaccinia Virus Ankara and play a key role in the induction of virus-specific T cell responses -3

N for IFN-α detection by ELISA. These data are representative of three independent experiments.<p><b>Copyright information:</b></p><p>Taken from "Dendritic Cells are preferentially targeted among hematolymphocytes by Modified Vaccinia Virus Ankara and play a key role in the induction of virus-specific T cell responses "</p><p>http://www.biomedcentral.com/1471-2172/9/15</p><p>BMC Immunology 2008;9():15-15.</p><p>Published online 15 Apr 2008</p><p>PMCID:PMC2359732.</p><p></p

Single and Repeated Low-Dose Challenge Designs

<p>Figure shows designs for single (A) and repeated (B) low-dose challenge designs. Small arrows denote challenges, and white and red symbols denote uninfected and infected animals, respectively.</p

Impact of Heterogeneity in Susceptibility on the Statistical Power

<div><p>(A) Susceptibility distributions for different levels of heterogeneity, measured by the coefficient of variation, <i>CV,</i> of the susceptibility distribution. The vaccine is assumed to be 80% effective, <i>VE_S =</i> 0.8.</p> <p>(B) The statistical power depends on the coefficient of variation, <i>CV,</i> for the repeated low-dose challenge design (black lines) and the single low-dose challenge design (green lines). For these plots we assumed trials with six and 12 animals per group and vaccine efficacies of <i>VE_S =</i> 0.67 (dotted lines), <i>VE_S =</i> 0.8 (dashed lines), and <i>VE_S =</i> 0.9 (solid lines).</p></div

Power Analysis for the Repeated Low-Dose Challenge Design and the Single Low-Dose Challenge Design

<p>In our virtual experiments, we set the challenge dose equal to the <i>ID</i>₅₀, and assumed that the vaccine efficacy was 67% (dotted lines), 80% (dashed lines), or 90% (solid lines). The graph shows the statistical power of the repeated low-dose challenge design (black lines) and the single low-dose challenge design (green lines) for a given number of animals per group as determined from 100,000 virtual experiments. If the vaccine is 90% effective, the statistical power of the repeated low-dose challenge design is higher than 95% with only five animals per group, as compared to only 15% for the single low-dose challenge design.</p

Impact of the Maximum Number of Challenges, <i>C_max,</i> on the Statistical Power

<p>For this plot we assumed trials with vaccine efficacies of <i>VE_S =</i> 0.67 (dotted line), <i>VE_S =</i> 0.8 (dashed line), and <i>VE_S =</i> 0.9 (solid line). In (A) we calculated the statistical power for six animals per group, <i>n =</i> 6, and in (B) for 12 animals per group, <i>n =</i> 12.</p

Evidence for Greater Positive Selection in SIV <i>env</i> at Later Times Postinfection

<div><p>(A–E) Calculations for dN and dS were performed along the 124 amino acids of the V1V2 region using SNAP (<a href="http://hiv-web.lanl.gov/" target="_blank">http://hiv-web.lanl.gov/</a>). The average dN and dS at each codon is shown for SMs at day 14 (A) and day 578 (B), as well as for RMs at day 14 (C) and day 578 (D), and for the SI (E). Yellow boxes indicate predicted N-gly sites, and asterisks indicate N-gly sites not present at early time points in RMs.</p><p>(F) Cumulative dN and dS are shown across all sites for each animal at day 14 and day 578. Raw values of cumulative dN and cumulative dS are indicated below the graph.</p></div