In vitro functionality of peripheral blood NK cells.

<p>NK cells from healthy individuals (n = 11) were stimulated for 6 hours with different concentration of RBV, PEG-IFNa and combination of both. (A) Representative FACS plots for NK cells stimulated with K562 target cells alone, with K562 target cells and RBV, with K562 target cells and PEG-IFNa, or with K562 target cells and combination of both are depicted. (B) Mean values of CD107a, IFNg and TNF expression on total NK cells upon stimulation with K562 target cells (B) and Huh7.5 hepatoma cells (C) from all healthy individuals.</p

In vitro functionality of peripheral blood NK cells.

<p>NK cells from healthy individuals (n = 11) were stimulated for 6 hours with different concentration of RBV, PEG-IFNa and combination of both. (A) Representative FACS plots for NK cells stimulated with K562 target cells alone, with K562 target cells and RBV, with K562 target cells and PEG-IFNa, or with K562 target cells and combination of both are depicted. (B) Mean values of CD107a, IFNg and TNF expression on total NK cells upon stimulation with K562 target cells (B) and Huh7.5 hepatoma cells (C) from all healthy individuals.</p

HCV RNA levels according to ART and CTM in samples obtained 4 weeks after start of TVR/BOC therapy that yielded levels ≥50 IU/ml in at least one of the two assays.

<p>The discordant results in two samples (“TVR 1” and “TVR 2”) would have led to different treatment decisions (treatment discontinuation based on HCV RNA levels>1000 IU/ml at week 4). In a single sample the ART measured a higher HCV RNA level than the CTM (“TVR 8”).</p><p>HCV RNA levels according to ART and CTM in samples obtained 4 weeks after start of TVR/BOC therapy that yielded levels ≥50 IU/ml in at least one of the two assays.</p

Phenotypical changes during treatment with RBV, PEG-IFNa and combination therapy.

<p>PBMC from HCV patients were stained directly ex vivo. Representative FACS plots for d0, d42 and d42 and single patient courses for (A) NKp30 (B) NKG2A and (C) CD57 expression on total NK cells are shown. Percentage of positive cells as well as fold change compared to baseline expression are represented.</p

Predictive value of HCV RNA results in the CTM and ART at week 4 (a) and week 8/12 (b) after start of therapy with a protease inhibitor.

<p>TND  =  target not detected; DET  =  HCV RNA detected; TND/TND  =  HCV not detected in both assays; TND/DET  =  HCV RNA detected by only one assay; DET/DET: HCV RNA detected by both assays. *Patients with unavailable virological treatment outcome were excluded from analysis.</p

NK cell function during treatment.

<p>PBMC from HCV patients were cultured in medium alone or in co-culture with K562 or Huh7.5 target cells without any further cytokine stimulation. (A) Representative FACS plots for CD107a, IFNg and TNF expression on total NK cells without stimulation (MED) and after stimulation with target cells are shown. (B) Mean values of CD107a, IFNg and TNF expression on total NK cells without stimulation (MED- grey line) or upon stimulation with target cells - K562 (red line) or Huh7.5 (blue line) are shown.</p

Frequency of NK cell subsets during therapy.

<p>(A) Gating strategy to distinguish NK cells and subgroups CD56^dim and CD56^bright cells. (B) Representative FACS plots for d0, d42 and d126 stainings and mean values for all patients are shown. d, day; RBV, Ribavirin; PLC, Placebo; PEG-IFN, pegylated Interferon.</p

HCV RNA levels according to ART and CTM in samples obtained 12 weeks after TVR treatment that yielded levels ≥50 IU/ml in at least one of the two assays.

<p>The discordant results in one sample (“TVR 3”) would have led to a different treatment decision (treatment discontinuation based on HCV RNA levels>1000 IU/ml at week 4).</p><p>HCV RNA levels according to ART and CTM in samples obtained 12 weeks after TVR treatment that yielded levels ≥50 IU/ml in at least one of the two assays.</p