Identification of cytokine profiles associated with ATB.

<p><b>A.</b> Unsupervised hierarchical clustering was performed according to the similarities in cytokine expression profiles which were visualised using the indicated color scale. Cytokine concentrations were indicated using a color scale that ranges from green (low) through black to red (high). The dendrogram above the heat map illustrated degrees of relatedness between the expression profiles evident within the various patients. Subjects with ATB (n = 35; black boxes on the strip above the heat-map), with high IFN-γ and low IL-12, TNF-α, GM-CSF, Eotaxin and IFN-γ secretions tended to cluster together (cluster I on the dendogram), while subjects with LTBI (n = 125; grey boxes) with IFN-γ and high IL-12, TNF-α, GM-CSF, Eotaxin and IFN-γ secretions also tended to cluster together (cluster II). The dendrogram on the right-hand side of the heat map indicates relationships between the expression profiles of the analysed cytokines across all the patients assessed in each study. <b>B.</b> Partial least squares analysis model of all 7 cytokines classified individuals with 61% overall accuracy for classification and 55% accuracy for cross-validation (red circles, ATB; green squares, LTBI). Latent Variable 1 (LV1), linear combination of the variables included in the model, LV2, orthogonal to the first combination. 95% of each population was circled. <b>C.</b> Latent variable 1 best separated individuals who are ATB from those LTBI. Cytokine loadings indicate multivariate cytokines associated with ATB.</p

Cytokines response in T-SPOT supernatants.

<p><b>A-</b> The expression of IFN-γ in ESAT-6 stimulated T cells was quantified by the T-SPOT assay. Results are expressed by numbers of secreted cells (SC) per 250 000 PBMC for 36 patients with ATB (red circles) and 115 patients with LTBI (green squares). The median is shown in each group. The p value is calculated by the Mann-Whitney U test. ** corresponds to p≤0.01. <b>B-</b> The ROC curve displays sensitivity vs specificity for IFN-γ in differentiating the ATB group from the LTBI group. The AUC and a threshold at 100 SC/250000 PBMC are indicated in the graph. <b>C-E-G-I-K-M-</b> IL-4, IL-12, TNF-α, GM-CSF, Eotaxin and IFN-α indexes (cytokine/IFN-γ *100), ATB are represented by red circles and LTBI by green squares. The median indexes are shown in each group of each panel. p values are calculated by the Mann-Whitney U test. ** corresponds to p≤0.01, *** to p≤0.001 and **** to p≤0.0001. <b>D-F-H-J-L-N-</b> Biparametric graphs between the numbers of IFN-γ immunospots in response to ESAT-6 stimulation and the level of cytokines (pg/ml) in the corresponding supernatants. ATB are represented by red circles and LTBI by green squares. Defined risk groups are circled.</p

Clinical characteristic of 150 subjects included in this study.

<p>Clinical characteristic of 150 subjects included in this study.</p

Design of crude odd ratios by univariate analysis depending on cytokine response from ESAT-6 stimulated PBMC.

<p>Design of crude odd ratios by univariate analysis depending on cytokine response from ESAT-6 stimulated PBMC.</p

Concentrations in pg/ml of cytokines secreted by PBMC after ESAT-6 stimulation in 36 patients with ATB and 115 with LTBI.

<p>LOD, limit of detection; n, number of values above detection level; nd, non determined.</p

Anti-Cut4 and anti-CFP21 memory B cells.

<p>(A) Anti-Cut4 and anti-CFP21 ASCs after polyclonal stimulation from representative subjects of the Mtb uninfected and LTB groups, 1.105 enriched B cells were seeded per well. Quantification of ASC after five days of culture, anti-Cut4 IgG (B) and anti-CFP21 IgG (C) in Mtb uninfected and LTB subjects. The median values have been drawn as horizontal lines. Values of Wilcoxon Mann Withney tests are indicated.</p

Serum antibody (IgG) levels against CFP21, Cut4 and PPD.

<p>The dot plots indicate level of antibodies directed against Cut4 (A), CFP21 (B), and PPD (C) in Mtb uninfected subjects, LTB subjects, and TB subjects. Antibodies levels are indicated per index of the optical density (OD), corresponding to the ratio of sample OD450nm/blank OD450nm.</p

Anti-Cut4 and anti-CFP21 antibody circulating secreting cells (ASCs).

<p>(A) Anti-Cut4 and anti-CFP21 ASCs from representative subjects of the three TB groups using ELISpot assay. 0.1 million enriched B cells were seeded per well. Quantification of cells secreting spontaneously anti-Cut4 IgG (B) and anti-CFP21 IgG (C) in Mtb uninfected subjects and subjects with LTB and active TB. The median values have been drawn as horizontal lines. Values of Wilcoxon Mann Withney tests are indicated.</p

Additional file 1: Figure S4. of Breastfeeding patterns and its determinants among mothers living with Human Immuno-deficiency Virus -1 in four African countries participating in the ANRS 12174 trial

Non-exclusive breastfeeding survival curves by country until week 50. This figure shows survival curves by country presenting the women nonexclusively breastfeeding their children during the 50-week follow-up period. (PDF 115 kb