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. 2014 Oct 2;10(10):e1004433. doi: 10.1371/journal.ppat.1004433

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(A) Left panel: Representative FACS plots showing the distribution of the monocyte subsets defined by CD14 and CD16 markers at week 0 and at week 6 or time of IRIS in IRIS vs. non-IRIS patients. Monocytes were also defined according to expression of CCR2 and CX3CR1 and overlaid onto CD14 vs. CD16 graphs. Right panel: Representative FACS plots showing histograms of various phenotypic markers in monocyte subsets based on CD14 and CD16 expression by clinical group and time point. (B) Percentage of circulating CD14⁺⁺CD16⁻ (left), CD14⁺CD16⁺ (center) and CD14^dimCD16⁺ (right) monocytes among circulating mononuclear myeloid cells (HLADR⁺CD2⁻CD3⁻CD19⁻CD20⁻CD56⁻) were compared at week 0 (pre-ART) and at week 6 or at the time of IRIS after ART initiation between TB-HIV co-infected patients that 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 analyses within each study group. (C) Representative FACS plots showing co-localization of CD163 and CD14 in circulating monocytes from an IRIS patient at week 2 of ART. (D) The median fluorescence intensity (MFI) of CD163 expression on CD14⁺⁺CD16⁻ monocytes is compared at week 0 (pre-ART) and week 6 or at the time of IRIS after ART initiation between TB-HIV co-infected patients that developed paradoxical TB-IRIS (n = 26) and those who did not (n = 22) using the Mann-Whitney test. (E) Relative risk (RR) of developing paradoxical TB-IRIS per standard deviation increase in the frequency of specific monocyte subsets at week 0 (pre-ART) after log₁₀ transformation. RR were adjusted for baseline age, gender, days to ART initiation, plasma HIV RNA levels and CD4⁺ T-cell count. CI, confidence interval. * P<0.05, ** P<0.01, *** P<0.001.