Cytokine/chemokine responses in TB cases and household contacts stratified by TST status after stimulation with ESAT^-6/CFP^-10 (EC) at enrolment.

<p>Following overnight or 6 day culture of peripheral blood mononuclear cells with EC, supernatants were collected and multiplex cytokine assays performed. Geometric mean levels are shown (pg/ml) A: Heat map showing geometric mean levels of different analytes after overnight (D1) versus 6 day (D6) culture in cases and contacts based on TST phenotype. The highest geometric means are shaded in red and the lowest in blue. Contacts were categorized according to TST scores at baseline and 6 months: TST⁺ = TST positive: TST≥10 mm at baseline; TSTC = TST converters: TST<10 mm at baseline and TST≥10 mm plus an increase in induration of at least 6 mm by 6 month; PTST⁻: persistently TST negative: TST<10 mm at both time–points. B: Heat map showing geometric mean levels of different analytes after overnight (D1) and 6 day (D6) culture in contacts grouped according to TST status and baseline EC IFN⁻γ ELISPOT (ECS) results. The highest geometric means are shaded in red and the lowest in blue.</p

Demographic, microbiologic and Mtb exposure characteristics of study participants.

<p><b>*</b> Fisher's exact test: p = 0.017 for proximity between TST⁺ and PTST⁻. ns = not significant; TST⁺ = TST positive; TSTC = TST converters; PTST⁻ = persistently TST negative; na = not assessed; IQR = interquartile range.</p><p>Demographic, microbiologic and Mtb exposure characteristics of study participants.</p

IFN–γ ELISPOT in cases and household contacts in response to stimulation with ESAT–6/CFP–10 (EC) or Purified Protein Derivative (PPD), at enrolment.

<p>Peripheral blood mononuclear cells (PBMC) from TB cases and contacts were stimulated overnight (A and B) or for 6 days (C and D) with EC (A and C) or PPD (B and D) and an IFN–γ ELISPOT was performed. Horizontal lines represent the geometric mean of the spot forming units (SFU). Contacts were categorized according to TST scores at baseline and 6 months: TST⁺ = TST positive: TST≥10 mm at baseline; TSTC = TST converters: TST<10 mm at baseline and TST≥10 mm plus an increase in induration of at least 6 mm by 6 month; PTST⁻: persistently TST negative: TST<10 mm at both time^-points. P^-values represent comparisons adjusted for household, sex and gender.</p

Mtb bacterial load pre- and post-treatment initiation.

<p>Sputum samples from patients pre and post treatment (n = 27) were analysed using the MBL assay. Line indicates median. Data were analysed using Wilcoxon matched pairs test.</p

Heatmap of cytokine profiles in patients with low and high bacterial loads.

<p>Cytokine levels (pg/ml) for unstimulated (NIL) and sputum supernatants were analysed using a 27-plex cytokine array. Subjects were grouped into those with low bacterial load (below the median of 48059 copies/ml) and high bacterial load (above median). Data were analysed using a Mann-Whitney U-test. NS = not significant. Colour indicates low (blue) to high (red) cytokine levels.</p

Correlation of MBL and host cytokines in blood and sputum.

<p>Bacterial load was analysed for each sputum sample and correlated with host biomarkers (pg/ml) in the same sputum sample and also in Mtb-stimulated and unstimulated blood using a 27-plex cytokine assay. R-values >0.4 are highlighted in yellow and significant p-values in purple. Data were analysed using a Spearman Rank Correlation. Correlation of Time-on treatment with host biomarkers in sputum was also performed.</p

Correlation of bacterial load with cycle threshold values.

<p>RT-PCR was performed in duplicate on RNA extracted from sputum samples of TB patients. <b>A:</b> Log₁₀ bacterial load versus Ct values of Mtb present in sputum pellets. <b>B:</b> Correlation of Ct values from GeneXpert and ¹⁶S RNA analysis. Data were analysed using Spearman rank correlation.</p

Patient demographics.

<p>Patient demographics.</p