Cytokines predictive of immunological set-point levels. (A–E)

<p>Cytokine concentrations in plasma at M0 have been plotted against CD4⁺ T cell counts and T cell activation (CD3⁺CD8⁺CD38⁺HLA-DR⁺) at M6. Six patients, including 4 who were treated at M6, were excluded from the analysis at M6. T cell activation levels were available for 19 patients at M6 (4 SP, 7 SP, 8 RP). The correlations were thus analyzed in 40 patients regarding CD4⁺ T cell counts and viral load and for 19 patients regarding T cell activation. (<b>A</b>) IP-10 levels at M0 plotted against T CD4⁺ counts at M6. (<b>B</b>) IP-10 levels at M0 plotted against T cell activation at M6. (<b>C</b>) IL-18 levels at M0 plotted against T cell activation at M6. (<b>D)</b> TGF-β1 concentrations at M0 plotted against T cell activation at M6. The red line indicates that both the Spearman correlation and the linear regression analysis were significant. <b>(E)</b> Regression analysis for evaluation of the capacity of CD4⁺ T cell counts, VL and cytokines at M0 to predict rapid disease progression. Values obtained for both the derivation set (Luminex and ELISA) and the validation set are shown. Median values of CD4⁺ T cell counts, VL and cytokine levels were used: VL > = 5 log; CD4<570 cells (derivation set); CD4<546 cells (validation set); IP-10> = 869 pg/ml (derivation set, Luminex); IP-10> = 247 pg/ml (derivation set, ELISA); IP-10> = 232 pg/ml (validation set, ELISA).</p

Plasma protein levels in HIV-1 infected patients.

<p>(<b>A</b>) The levels of 28 proteins in the plasma of 46 acutely infected patients (M0) are expressed as fold change compared to the levels in healthy donors (N = 17). The order of the proteins is presented according to their function (pro-inflammatory, adaptive, IFN-inducible, chemoattractants, hematopoietic and anti-inflammatory). The anti-inflammatory cytokines are presented on the right side of the figure. The dotted horizontal line at Y = 1 corresponds to the value in healthy donors. The boxes represent the median and the 25^th and 75^th percentile, with the line in the middle of the boxes corresponding to the median value. Colored boxes stand for the cytokines, whose levels were significantly different from healthy donors: blue boxes when p<0.05 (as it was the case for IL-1β) and red boxes when p<0.008 (M&W test). (<b>B</b>) Protein concentrations at M0, M1 and M6 of the soluble factors that were elevated at M0. The data are expressed in pg/ml, HD: healthy donors, M: months. Cytokine concentrations below the limit of detection were arbitrarily set at the level of the limit of detection. Dot-plots marked with one asterisk (*) (p<0.05) or two asteriks (**) (p<0.008) represent the cytokines, whose levels were significantly different from healthy donors (M&W test).</p

Comparative distribution of patient’s demographic, clinical and immunological characteristics in the derivation and validation sets.

<p>The main characteristics of the patients are listed here for the time point of enrollment during PHI (M0). For each parameter (except gender), the median values and the range are indicated. Plasma IP-10 concentrations were measured by ELISA at M0. For the derivation set, data are shown for those 45 patients out of 46, whose IP-10 was quantified by ELISA. The validation set comprised 88 patients. There was no significant difference between the two sets for any parameter (p>0.11). W: women, M: men, N: number of patients in each set.</p

Plasma protein levels at M0 according to disease progression profiles.

<p>The cytokine profiles at M0 are shown for each group of patients: 16 rapid progressors (<b>A</b>), 19 progressors (<b>B</b>), and 11 slow progressors (<b>C</b>). Color code and statistical analyses are as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0046143#pone-0046143-g002" target="_blank">Figure 2</a> (corrected threshold, p<0.005). The dotted horizontal line corresponds to the value in healthy donors. (<b>D</b>) Comparison of protein concentrations between the 3 groups of patients (SP, P and RP). Four representative cytokines are shown. The cytokines increasing significantly over groups were IP-10 and IL-10 (Cuzick’s test, p<0.007). When comparing the groups two by two, out of 28 proteins tested, the levels were different only for IP-10 (M&W test,***: p<0.005).</p

Immunological, virological and clinical characteristics of rapid progressors versus progressors and slow progressors assembled within patients of the derivation and validation sets.

<p>The derivation set comprised 46 patients (except for IP-10, where values based on the ELISA were available for only 45 patients). The validation set corresponded to 88 patients for all markers. Differences between rapid progressors and the other patients are indicated with the presence of a p value (M&W test). P values below 0.05 were considered to be significant. There was no significant difference regarding the time of enrollment at M0 (estimated days post-infection). (A) CD4⁺ T cells at M0 and M12; (B) Viremia at M0 and M12; (C) number of estimated days post-infection at M0; (D) Plasma IP-10 concentration at M0. M = month.</p