Association of IFNγ to <i>M.tb</i> PPD with 25(OH)D concentrations 3 months post BCG vaccination.

<p>25 (OH) D was measured in plasma samples and IFNγ to <i>M.tb</i> PPD was measured by ELISA from supernatants from 6 day cultures of diluted whole blood from infants 3 months post BCG vaccination. <b>a</b>] Dot plot where individuals are represented by dots, and lines represents linear fit of responses <b>b</b>] Histograms of IFNγ response to <i>M.tb</i> PPD response in infants with hypovitaminosis (<30 ng/ml) and in infants with sufficient 25(OH)D.</p

Infants vitamin D status at time-point 2 by infant characteristics, with groups compared according to mean difference and crude odds ratios for no vitamin D deficiency (>20 ng/ml).

<p>Severe Vitamin D Deficiency (sVDD), mild Vitamin D Deficiency (mVDD), Vitamin D Insufficiency (VDI) and Vitamin D Sufficiency (VDS).</p><p>**LR =  likelihood ratio test.</p><p>***1 data point missing as ethnic group “not stated”.</p

Baseline proportions and association of factors with default by univariate analysis.

<p>BMI = body-mass index. MDR = multidrug-resistant. Cat I = category I treatment. Cat II = category II treatment. XDR = extensively drug-resistant.</p

Histograms of 25(OH)D at time-point 1 and 2.

<p>25(OH)D was measured in plasma samples from infants 3 and 12 months post BCG vaccination, and in age-matched unvaccinated controls. 25(OH)D in BCG vaccinated and unvaccinated infants <b>a</b>] in vaccinated infants and unvaccinated controls combined at time-point 1 and <b>b</b>] in vaccinated infants and unvaccinated controls combined at time-point 2; 25(OH)D in infants grouped according to BCG vaccination status at <b>c</b>] time-point 1 and <b>d</b>] time-point 2.</p

Factors associated with default identified by multivariate analysis.

<p>OR = odds ratio. XDR = extensively drug-resistant. Cat II = category II treatment.</p

Infant vitamin D concentrations at time-point 1, 3 months post-BCG vaccination compared to age-matched controls: mean differences and odds ratios for the effect of BCG vaccination, stratified on and controlled for season of blood collection, ethnic group and sex.

<p>*unadjusted (“crude”),</p><p>**adjusted.</p

Infant vitamin D status at time-point 1, by infant characteristics, with groups compared according to mean differences and crude odds ratios for Vitamin D Sufficiency.

<p>*severe Vitamin D Deficiency (sVDD), mild Vitamin D Deficiency (mVDD), Vitamin D Insufficiency (VDI) and Vitamin D Sufficiency (VDS).</p><p>**LR =  likelihood ratio test.</p><p>***1 data point missing as ethnic group “not stated”.</p

The proportion of cytokine responders to PPD at baseline and follow-up time points.

<p>The proportion of responders producing IFN-γ (A), IL-13 (B), IL-10 (C), CXCL10 (D), IL-17 (E) and TNF-α (F) in response to PPD was analysed in 40 spouses at baseline, 49 at 6 months, 39 at 12 months and 22 at 24 months follow-up. The majority of the spouses did not produce IL-13 and IL-10 in response to PPD at baseline or 24 months later while high levels of IFN-γ, CXCL10, IL-17 and TNF-α were detected. There were no significant changes in the proportions of cytokine responders throughout the follow-up period.</p

The proportion of cytokine responders to ESAT-6 at baseline and follow-up time points.

<p>The proportion of responders producing IFN-γ (A), IL-13 (B), IL-10 (C), CXCL10 (D), IL-17 (E) and TNF-α (F) in response to ESAT-6 was analysed in 40 spouses at baseline, 49 at 6 months, 39 at 12 months and 22 at 24 months follow-up. There were no significant differences in the proportions of IFN-γ responders between baseline and 24 months follow-up. Most of spouses did not produce IL-13 in responses to ESAT-6 at baseline, and this did not change over the 24 months follow-up. The proportions of IL-10 and TNF-α responders to ESAT-6 gradually declined at 6, 12 and 24 months.</p

The ratio of cytokine responses in spouses and TB patients.

<p>The ratio of cytokine responses to ESAT-6 (A) and PPD (B) were analysed in the spouses who showed negative IFN-γ responses to ESAT-6 (<62.5 pg/mL, n = 102; indicated as S1), the spouses with positive IFN-γ responses (>62.5 pg/mL, n = 48; indicated as S2) and TB patients (n = 15; indicated as T). The ratio of IFN-γ/IL-10, CXCL10/IL-10, TNF-α/IL-17, IFN-γ/IL-17 and IL-17/IL-10 differentiated the S2 and TB groups from S1 in responses to both ESAT-6 and PPD. None of the ratios differentiated between the S2 and T groups upon ESAT-6 stimulation while IFN-γ/IL-10 and IFN-γ/IL-17 in response to PPD distinguished S2 from TB patients. Median ratio of cytokine responses is marked with horizontal bars. (* P<0.05, ** P<0.01, *** P<0.001)</p