Number of Patients Alive, Assessable, and Dead at Each Study Point

<p>Data are from the French prospective multicentre CryptoA/D study. Pts, patients.</p

Vaccination status according to A/H1N1 vaccination status.

<p>Vaccination status according to A/H1N1 vaccination status.</p

Measures of personal prevention.

<p>Measures of personal prevention.</p

Published series (1982–2007) of more than 5 HIV-infected adult cases on radiological presentation of cerebral cryptococcosis.

a<p>CT: computed tomography.</p>b<p>MR: magnetic resonance image.</p

ROS induction by Thimerosal in TCR-activated T cells and anti-apoptotic effect of NAC.

<p>A- PBMC from a HD were stimulated overnight with anti-CD3 mAbs in the presence of thimerosal at 3 μg/ml, and NAC (2.5 μM) was added in half of the cultures. ROS production was detected with HE, ΔY_m was assessed using the DiOC₆(3) probe, and apoptosis was measured with 7-AAD. The dot plots show combined analysis of ΔY_m and apoptosis or ΔY_m and ROS production on gated CD4⁺ T cells, in a representative experiment out of three experiments performed with three different HD. B- PBMC from a HD were either unstimulated (left panel) or stimulated overnight with anti-CD3 mAbs in the presence or absence of thimerosal at 3 μg/ml (right panel). The anti-oxidant NAC (2.5 μM) was added in some cultures. The percentages of CD4⁺ and CD4⁻ T cells with a concomitant drop in ΔY_m and increased oxidizing ability (DIOC₆^high 7-AAD^neg) under the indicated culture conditions are shown. The bars indicate the mean and standard deviation from 3 experiments with three different donors. Asterisks indicate significant <i>P-</i>values (** <0.001, ***0.0001) for comparison of cultures stimulated with thimerosal and NAC to cultures stimulated with thimerosal only.</p

Neuroradiological analysis of 55 computed tomographies and 24 magnetic resonance images collected at baseline from 62 HIV-infected patients with culture-proven cryptococcal meningoencephalitis.

a<p>With evidence of subdural collection.</p>b<p>Some patients had more than one lesion.</p

Impact of thimerosal on cytokine and chemokine release upon TCR ligation.

<p>Pattern of cytokines and chemokines simultaneously quantified by multiplex bead assay arrays in supernatants from freshly isolated PBMC following 16-hour stimulation with anti-CD3/CD28 mAbs in the absence or presence of three different concentrations of thimerosal. The bars indicate the mean and standard deviation from experiments with three different donors. Asterisks indicate significant <i>P-</i>values (* <0.05, ANOVA test) for comparison with cultures without thimerosal.</p

Examples of abnormal radiological findings.

<p>A. Magnetic Resonance axial T2-weighted image, displaying bilateral dilated Virchow-Robin spaces (arrow) in the basal ganglia. B. Magnetic Resonance axial T2-weighted image displaying a hyperintense right occipital mass (arrow head) and bilateral dilated Virchow-Robin spaces (arrow). C. Magnetic Resonance axial T1-weighted image with contrast infusion displaying frontal subdural collection (arrow). D. Magnetic Resonance axial T1-weighted image with contrast infusion displaying a basal meningeal enhancement (arrow).</p

Dose-dependent Toxicity of Panenza on Memory T cells specific for influenza vaccine.

<p>A- PBMC from patients vaccinated with the seasonal flu vaccine (Mutagrip) and the pandemic 2009 H1N1 vaccine (Pandemrix) were collected 21 days after vaccine administration. Cells (5×10⁵ per well) were stimulated for 5 days with Mutagrip or the nonadjuvanted pandemic 2009 H1N1 Panenza vaccine at the indicated concentrations (vaccine concentration is expressed as the final concentration of HA). Cells were also stimulated with CMV or EBV peptides at 0.25 μg/ml, tetanus toxoid (TT) or tuberculin PPD at 5 μg/ml. Cultures were pulsed with 1 μCi per well of [3H] thymidine over the final 16 h of culture and cell proliferation expressed in cpm. All tests were done in triplicate and results show the mean values for three patients. The bars indicate the mean and standard deviation. Asterisks indicate significant <i>P-</i>values (p = 0.03) for comparison of cultures stimulated with Panenza at 0.02 to 0.5 μg/ml to cultures stimulated with 0.01 μg/ml of Panenza (ANOVA test). B- PBMC from Pandemrix vaccinated patients were incubated with soluble Panenza at indicated concentrations, or added to plates coated overnight at 4°C with indicated concentrations of Panenza. H3TdR (1μCi/well) was added at day 4 and T-cell proliferation was assessed at day 5. The bars indicate the mean and standard deviation. Asterisks indicate significant <i>P-</i>values (* <0.05, ** <0.01, ***<0.001) for comparison of cultures stimulated with coated Panenza to cultures stimulated with soluble Panenza. C- PBMC from a patient vaccinated with Mutagrip and then with Pandemrix were incubated overnight with culture medium, Mutagrip or Panenza at 0.25 μg/ml and the percentage of apoptotic cells was determined by flow cytometry combining 7-AAD staining with CD3 and CD4 membrane detection. Dot plots represent the co-expression of CD4 and 7-AAD on gated CD3⁺ T cells. D- PBMC from a patient vaccinated with Mutagrip and Panenza were incubated overnight with either vaccine at various concentrations and the percentage of apoptotic (7-AAD⁺) cells within indicated subsets was determined combining 7-AAD staining with CD19, CD3, CD8, CD4, CD56, CD16 or CD14. B cells were CD3⁻CD19⁺, T cells were CD3⁺CD8⁺ or CD3⁺ CD4⁺, NK cells were CD3⁻CD56⁺CD16⁺, monocytes were CD3⁻CD19⁻CD14⁺.</p

Thimerosal induces cytochrome c release and caspase-3 activation.

<p>A- Confocal analysis of PBMC incubated 16 hours either in the presence of thimerosal (0.9 or 3 μg/ml), or with staurosporine (1 μg/ml) as a positive control. Cells were processed for cytochrome c staining (green) and co-stained with DAPI (blue) to detect nuclei modifications. B-C Western blot analysis of active caspase-3 expression in PBMC incubated for 16 h in medium (unstimulated) (B) or stimulated with anti-CD3 mAbs (C). The impact of indicated concentrations of thimerosal or staurosporine on active caspase-3 expression is shown, together with the influence of the broad caspase inhibitor QVD and the anti-oxidant NAC.</p