Characteristics of the patients IRIS and non-IRIS at pre-ART.

<p>Characteristics of the patients IRIS and non-IRIS at pre-ART.</p

Associations between pre-ART characteristics and occurrence of severe IRIS.

<p>Associations between pre-ART characteristics and occurrence of severe IRIS.</p

Comparison between severe and non-severe cases of patients in the TB-IRIS study.

<p>Comparison between severe and non-severe cases of patients in the TB-IRIS study.</p

Pre-ART plasma levels of IL-6 or CRP distiguish individuals at higher risk for TB-IRIS.

<p>(A) Pre-ART plasma concentrations of IL-6 (left panel) and CRP (right panel) were compared between individuals who developed TB-IRIS during study follow up and those who did not using the Mann-Whitney test. (B) Correlation between IL-6 and CRP values was tested in both groups of patients using the Spearman test. In (B), dotted lines represent the median pre-ART values of IL-6 or CRP in the entire study population. Gray areas highlight those patients displaying values for both biomarkers above their respective median values within the study population. (C) The associations between systemic levels of IL-6 and CRP with risk for subsequent TB-IRIS were assessed by univariate and multivariate models. Relative risks (RR) are per standard deviation increase after log₁₀ transformation. RR were adjusted for baseline age, gender, weight, hemoglobin, hematocrit, sputum culture grade, presence of extra pulmonary TB, presence of miliary TB, days to ART initiation, plasma HIV RNA levels and CD4⁺ T-cell count. CI, confidence interval.</p

Baseline characteristics of the study participants.

<p><b>NOTE.</b> Data represent no. (%) of participants unless otherwise specified. ALT, alanine transaminase; AST, aspartate aminotransferase; ATT, anti-tuberculous treatment; EPTB, extra-pulmonary tuberculosis; IQR, interquartile range; RBC, red blood cell; TB, tuberculosis.</p>*<p>The ATT regimens differentiated according to the frequency of drug administration (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0063541#s2" target="_blank">Methods</a> for full discrimination of the regimens).</p

Baseline characteristics of the study population stratified by IRIS status.

<p><b>NOTE.</b> Data represent no. (%) of participants unless otherwise specified. ALT, alanine transaminase; AST, aspartate aminotransferase; ATT, anti-tuberculous treatment; EPTB, extra-pulmonary tuberculosis; IQR, interquartile range; RBC, red blood cell; TB, tuberculosis.</p

Associations between pre-ART clinical and laboratory characteristics with subsequent TB-IRIS events.

<p>Baseline (pre-ART) characteristics significantly different between patients that developed TB-IRIS events in the follow up and those who did not were assessed to test associations with risk for IRIS in univariate and multinomial logistic models. Relative risks (RR) are for values below or above the threshold levels displayed, which were estimated close to median values for the overall study population. Adjustment was performed for all variables presented and also included age and gender. 95% CI, 95% confidence interval.</p

Circulating monocyte counts do not correlate with soluble biomarkers of inflammation and immune activation in TB-HIV co-infected individuals.

<p>(<b>A</b>) Numbers of circulating monocytes (left panel) and neutrophils (right panel) are compared at week 0 (pre-ART), at weeks 6 or at the time of IRIS, and at week 24 after ART initiation between TB-HIV co-infected patients who developed paradoxical TB-IRIS (n = 26) and those who did not (n = 22). Lines represent median values and interquartile ranges. Data were analyzed using the Mann-Whitney test or Wilcoxon matched-pairs test for paired comparisons within each study group. ** P<0.01, *** P<0.001. (<b>B</b>) The network analysis (interactome) showed statistically significant correlations (P<0.05) between neutrophil or monocyte counts and plasma biomarkers. Associations were assessed with Spearman rank tests.</p

Expression profile of plasma biomarkers of inflammation and immune activation in TB-HIV co-infected patients on anti-TB therapy.

<p>(<b>A</b>) Left panel shows <i>Mycobacterium tuberculosis</i> sputum culture grade in samples from TB-HIV co-infected individuals at study enrollment (pre-ART) stratified by time on anti-TB treatment (ATT). Data were compared using the Mann-Whitney test. Right panel shows the frequency of IRIS vs. non-IRIS patients taking ATT for different durations (≤4 weeks and>4 weeks). Data were compared using the Chi-square test. (<b>B</b>) A heat map was designed to depict the overall pattern of expression of plasma cytokines, chemokines and inflammatory biomarkers in TB-HIV co-infected patients at different time points after ATT initiation. A hierarchical cluster analysis (Ward's method) of circulating biomarkers by clinical group and time point was performed. Expression scale for each biomarker represents change from the geometric mean of the entire study population (n = 48). Differences between medians were compared using the Mann-Whitney test. * P<0.05, ** P<0.01, *** P<0.001.</p

Plasma concentrations of monocyte activation markers are increased during TB-IRIS and correlate with markers of systemic inflammation.

<p>(<b>A–C</b>) Plasma levels of soluble (s) CD14 (<b>A</b>), sCD163 (<b>B</b>) and sTF (<b>C</b>) are compared at week 0 (pre-ART), at week 6 or at the time of IRIS, and at week 24 after ART initiation between TB-HIV co-infected patients who developed paradoxical TB-IRIS (n = 26) and those who did not (n = 22). Lines represent median values and interquartile ranges. Data were analyzed using Mann-Whitney or Wilcoxon matched-pairs test for paired analyses within each study group. * P<0.05, ** P<0.01, *** P<0.001. (<b>D</b>) Left panel: A heat map was designed to depict the overall pattern of expression of plasma cytokines, chemokines and inflammatory markers in patients developing TB-IRIS vs. non-IRIS controls at week 0 (pre-ART) and at week 6 or during the time of IRIS. A two-way hierarchical cluster analysis (Ward's method) of circulating biomarkers by clinical group and time point was performed. Expression scale for each biomarker represents log₁₀ fold-change from the geometric mean of the entire study population at week 0 and week 6 or time of IRIS (n = 48 in each study time point). Right panel: The network analysis (interactome) shows statistically significant correlations (P<0.05) between all the variables measured. Data were analyzed using Spearman rank tests. See <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1004433#ppat.1004433.s001" target="_blank">Data File S1</a> for additional details on the strength (r value) and level of significance (P-value) of each individual correlation.</p