Sensitivity and specificity of the Xpert Ebola assay versus the Trombley assay performed on clinical whole blood and buccal swab samples.

<p>Sensitivity and specificity of the Xpert Ebola assay versus the Trombley assay performed on clinical whole blood and buccal swab samples.</p

Comparison of cycle threshold values for whole blood and buccal swab specimens that tested positive by Trombley and/or Xpert assay.

<p>Comparison of cycle threshold values for whole blood and buccal swab specimens that tested positive by Trombley and/or Xpert assay.</p

Evaluation of cycle threshold value agreement.

<p>Bland—Altman plots to evaluate agreement between cycle threshold values generated for the NP target by the Trombley versus Xpert assay for WB (A) and BS (C) samples and those generated by Xpert for the NP versus GP target for WB (B) and BS (D) samples. The blue line in each plot represents the “bias,” or average difference between the two methods. The red lines represent the 95% limits of agreement. The green line represents the line of no bias.</p

A Comparison of the Adaptive Immune Response between Recovered Anthrax Patients and Individuals Receiving Three Different Anthrax Vaccines

<div><p>Several different human vaccines are available to protect against anthrax. We compared the human adaptive immune responses generated by three different anthrax vaccines or by previous exposure to cutaneous anthrax. Adaptive immunity was measured by ELISPOT to count cells that produce interferon (IFN)-γ in response to restimulation <i>ex vivo</i> with the anthrax toxin components PA, LF and EF and by measuring circulating IgG specific to these antigens. Neutralising activity of antisera against anthrax toxin was also assayed. We found that the different exposures to anthrax antigens promoted varying immune responses. Cutaneous anthrax promoted strong IFN-γ responses to all three antigens and antibody responses to PA and LF. The American AVA and Russian LAAV vaccines induced antibody responses to PA only. The British AVP vaccine produced IFN-γ responses to EF and antibody responses to all three antigens. Anti-PA (in AVA and LAAV vaccinees) or anti-LF (in AVP vaccinees) antibody titres correlated with toxin neutralisation activities. Our study is the first to compare all three vaccines in humans and show the diversity of responses against anthrax antigens.</p></div

The characteristics of individuals enrolled into this study including: age, number of exposures to <i>B</i>. <i>anthracis</i> antigens and time since exposure in relation to the type of exposure to <i>B</i>. <i>anthracis</i> antigens.

<p>Panel A shows the distribution of age across groups in study including individuals with no history of Anthrax, recent Anthrax infection, or one of three different vaccines. Each symbol denotes a single participant, the line is the mean and the error bars 95% Confidence Intervals. Statistical analysis is shown where Levene’s test validated GLM has been performed with Bonferroni’s post-tests. Panel B shows the distribution of time since exposure to vaccine or onset of anthrax symptoms across groups in the study. Each symbol denotes a single participant, the line is the median and the error bars the quartile range. The data were analysed by Kruskal-Wallis test (excluding naïve controls), which indicated differences between groups with Dunn’s individual comparisons. Panel C shows the distribution of number of anthrax-based immunological stimuli (doses) across groups in the study including percent people who had had Anthrax infection, or one of three different vaccines. Each symbol denotes a single participant, the line is the median and the error bars the quartile range. The data (excluding naïve and naturally infected controls) were analysed by Kruskal-Wallis test, which indicated differences between groups with Dunn’s individual comparisons. Significance markers indicate either P-values associated with either Bonferroni’s (For GLM) or Dunn’s (for Kruskal-Wallis) multiple comparisons where * indicates P < 0.05, ** indicates P < 0.01 and *** indicates P < 0.001).</p

Correlations between immune response to <i>B</i>. <i>anthracis</i> antigens and individual’s age and the number of vaccine doses.

<p>Panel A shows the correlation between age and antibody response to EF, in participants of a study which included individuals who had suffered from cutaneous anthrax previously, been vaccinated with either AVA, LAAV or AVP, or have no history of exposure to anthrax antigens. Antibody responses were measured by ELISA estimating antigen specific antibody in μg / ml. Panels B-D show the correlation between the numbers of doses of vaccine received and antibody response. Antibody responses were measured by ELISA estimating antigen specific antibody against PA (panel B), LF (panel C) and toxin neutralisation activity (Panel D). Partial correlation analyses indicated a significant negative correlation between the parameters (marked on the graphs).</p

The memory immune responses towards parts of the Anthrax toxin systems in individuals who had suffered from cutaneous anthrax previously, been vaccinated against anthrax with either AVA, LAAV or AVP, or have no history of exposure to anthrax antigens.

<p>Antibody responses were measured by ELISA, estimating antigen specific antibody in μg / ml or a dilution end point. IFN-γ T cell responses were estimated by ELISPOT and numerated as fold induction in IFN-γ producing cells by exposure to anthrax antigen, when compared to unstimulated cells. Toxin neutralisation was measured in the ED₅₀ of cells exposed to anthrax toxin in the presence of the serum. ULQ signifies the upper limit of quantification, due to confluence of spots on the ELISPOT plate, which was regarded as 25 fold. An additional line has been included which is indicative of the 90 percentile of the naïve group. The purpose of this is to give an approximate indication of the number of individuals with greater than background antigen recognition. Statistical analysis is shown. First, Levene’s tests of unequal variances were performed to establish suitability for parametric analysis. Unsuitability necessitated Kruskal-Wallis tests with Dunn’s post-tests. Suitability allowed for GLM analysis which included any potential covariates suggested previously, with Bonferroni’s multiple comparisons. The significance markers show results post-test comparisons: *** = P < 0.001, ** = P <0.01 * = P <0.05.</p

Frequency of gender between groups in study including individuals with no history of Anthrax, recent Anthrax infection, or one of three different vaccines.

<p>Frequency of gender between groups in study including individuals with no history of Anthrax, recent Anthrax infection, or one of three different vaccines.</p

The relativity of different antigen specific responses in groups exposed to <i>B</i>. <i>anthracis</i> antigens in different ways.

<p>The panels show the five different groups investigated in the course of this study. Participants included individuals who had suffered from cutaneous anthrax previously, been vaccinated with either AVA, LAAV or AVP, or had no history of exposure to anthrax antigens. Each line represents an individual with 7 different immune measurements take. Antibody responses were measured by ELISA estimating antigen specific antibody in μg / ml or a dilution end point. IFN-γ T cell responses were estimated by ELISPOT and numerated as fold induction in IFN-γ producing cells by exposure to anthrax antigen, when compared to unstimulated cells. Toxin neutralisation was measured in the ED₅₀ of cells exposed to anthrax toxin in the presence of the serum. The analyses are Pearson Correlations and the R values are given with the significance markers of *** = P < 0.001, ** = P <0.01 * = P <0.05.</p

Survival plot with futility boundary for TKM-130803 recipients.

<p>The red line denotes the futility boundary. The points and dashed line denote the number of survivors at day 14 plotted against the number of day 14 reports.</p