IFNα therapy leads to major impairment of thymic function.

<p>(A) The frequency of Ki-67 expressing cells in the CD4⁺ RTE subset (CD31^hi naïve T-cells) was measured in acutely HCV-infected (grey symbols, top panel), chronically HCV-infected (white symbols, top panel) and HIV/HCV co-infected (bottom panel) patients (central panels) and HIV/HCV co-infected (right panels) patients. Each line represents data from an individual patient. Statistical significances of the differences to baseline values (time 0), calculated on the absolute sj/βTREC ratio in each individual sample (Wilcoxon matched-pairs signed-ranks test) are shown on top. The horizontal bars represent median values.</p

IFNα therapy leads to reduction in IL-7 plasma concentration.

<p>(A) IL-7 plasma levels were quantified in peripheral blood cells from acutely HCV-infected (light grey symbols), chronically HCV-infected (black symbols) and HIV/HCV co-infected (white symbols) patients at study entry, as compared to healthy donors (HCV-, dark grey symbols). **: p<0.001 for any HCV-infected patients group. (B) Evolution of plasma IL-7 levels over the first 4 months of IFNα therapy in acutely HCV-infected (left panels), chronically HCV-infected (central panels) and HIV/HCV co-infected (right panels) patients. Each line represents an individual patient. Statistical significances of the differences to baseline values (time 0), calculated on the absolute IL-7 plasma levels in each individual sample (Wilcoxon matched-pairs signed-ranks test) are shown on top. (C) Soluble CD127 was quantified in plasma from acutely HCV-infected (white symbols, top panel), chronically HCV-infected (black symbols, top panel) and HIV/HCV co-infected (bottom panel) patients at baseline (0) and M2. (D) CD127 expression was measured on circulating CD4⁺ (top panel) and CD8⁺ (bottom panel) and expressed as mean fluorescence intensity (left panels) and percentages of positive cells (right panels) over the 4 first months of IFNα therapy.</p

IFNα therapy leads to naïve T-cell lymphocytopenia.

<p>(A) CD4⁺ (top panel) and CD8⁺ (bottom panel) naïve T-cell counts were quantified in peripheral blood cells from acutely HCV-infected (light grey symbols), chronically HCV-infected (black symbols) and HIV/HCV co-infected (white symbols) patients at study entry, as compared to healthy donors (HCV-, dark grey symbols). (B) Evolution of CD4⁺ (top panels) and CD8⁺ (bottom panels) naïve T-cell counts during the first 4 months of IFNα therapy in acutely HCV-infected (left panels), chronically HCV-infected (central panels) and HIV/HCV co-infected (right panels) patients. Each line represents data from an individual patient. Statistical significances of the differences to baseline values (time 0), calculated on the absolute naïve T-cell counts in each individual sample, (Wilcoxon matched-pairs signed-ranks test) are shown on top. The horizontal bars represent median values.</p

Variations in IL-7 plasma levels correlate with evolution of RTE

<p><b>production.</b> Correlations. between variations in IL-7 plasma levels (ΔIL-7) and either variations in (A) total (CD4⁺ + CD8⁺) naïve T-cell counts (Δnaïve T-cell counts), (B) RTE defined as CD31^hi naïve CD4⁺ T-cells (ΔRTE CD4 counts), (C) the sj/βTREC ratio (Δsj/βTREC ratio), (D) the frequency of Ki-67⁺ cells in the RTE CD4⁺ T-cell subset (Δ%Ki-67⁺ in CD4⁺ RTEs) or (E) the number of circulating Ki-67⁺CD4⁺ RTEs (ΔKi-67⁺ RTE counts) between study entry and month 1 of therapy were calculated for acutely (black symbols) and chronically (white symbols) HCV-infected patients (left panels) and HIV/HCV co-infected patients (right panels). Correlation coefficients (Spearman's r) and the associated probabilities (p) are shown.</p

IgA autoantibody profile.

<p>Heatmaps of serum IgA autoantibody reactivity in adults thymectomized early in infancy (T) and controls (C), clustered by autoantigen and subject group. Reactivity intensities were normalized and log2-transformed; and 121 IgA autoantibodies meeting minimal net fluorescence intensity requirements are presented. Key color bar corresponds to quantified reactivity.</p

IgM autoantibody profile.

<p>Heatmaps of serum IgM autoantibody reactivity in adults thymectomized early in infancy (T) and controls (C), clustered by autoantigen and subject group. Reactivity intensities were normalized and log2-transformed; and 122 IgM autoantibodies meeting minimal net fluorescence intensity requirements are presented. Key color bar corresponds to quantified reactivity.</p

Macaque bone marrow pDC express lower CD123 and HLA-DR, display higher percentages of CD34+ and Ki67+ precursors than blood pDC, and are poor IFN-I producers.

<p>(<b>A</b>) CD34⁺ and Ki67⁺ pDC-precursor frequencies are higher in the bone marrow (BM) than in the blood (non-infected macaques, n = 6), and both CD123 and HLA-DR expressions are lower in BM pDC than blood pDC. From <b>left to right</b>: Percentage of CD34⁺ pDC precursors, percentage of Ki67⁺ pDC precursors, and CD123 and HLA-DR geometric MFI (gMFI) in BM and blood pDC. Dotplot showing Ki67 and CD34 expression by pDC in BM and blood, from one representative animal. (<b>B</b>) Bone marrow pDC produce less IFN-I in response to TLR-7/8 stimulation (R848) than blood pDC (non-infected macaques, n = 6) (<b>Left</b>). Most IFNα is produced by Ki67⁻ pDC and not by K67⁺ precursors (<b>Middle</b>). Representative dot plot showing that only Ki67⁻ pDC produce IFNα (<b>Right</b>). Wilcoxon's rank sum test was used for all comparisons of paired data.</p

IgE autoantibody profiles.

<p>Heatmaps of serum IgE autoantibody reactivities in adults thymectomized early in infancy (T) and controls (C) clustered by autoantigen and subject group. Reactivity intensities were normalized and log2-transformed, and 100 IgE autoantibodies meeting minimal net fluorescence intensity requirements are presented. Key color bars correspond to quantified reactivity.</p

Clinical, epidemiologic and immunologic characterization.

<p>Clinical, epidemiologic and immunologic characterization.</p

Plasmacytoid DC produce IFNα in both lymphoid and mucosal compartments.

<p>Dotplot showing IFNα intra-cellular staining in gated pDC (CD45⁺HLA-DR⁺lin⁻CD123⁺) in different tissues. Cells were labeled <i>ex vivo</i> on fresh cells after 30 min incubation in 10 µg/mL brefeldin A in the absence of any stimulation. Data for two macaques sacrificed on day 10 p.i. and one uninfected control are shown. Mononuclear cells from BM, spleen, peripheral LN, mesenteric LN, ileum, and colon were extracted for FACS analysis. Frequencies of IFNα-pDC are indicated in bold and # indicates the number of pDC recorded for each file.</p