Peripheral blood mononuclear cell phenotype characteristics in different study groups before antiviral treatment.

<p>The distribution of peripheral blood lymphocyte subsets was similar between study groups. Absolute monocyte count was significantly increased in rapid virological responders compared to early and non-responder patients. Results are expressed as mean±SE.</p>*<p>p<0,05.</p

The effect of PEG-IFN plus ribavirin treatment on Th1/Th2 cytokine production by PMA/Ionomycin stimulated PBMC.

<p>a. IFN-γ production was significantly increased in complete early virological responders during antiviral treatment compared to pretreatment levels and also to null-responders. b. After 4 and 12 weeks of antiviral treatment, a transient increase in IL-2 production was observed in all study groups. c. PEG-IFN plus RBV resulted in decreased IL-6 production in both rapid and complete early virological responders, had a transient effect in null-responders. d. After 24 weeks of treatment, significantly decreased TNF-α production was found in complete early virological responders. e. f. While PEG-IFN/RBV treatment significantly decreased IL-4 and IL-10 levels in complete early virological responders, null-responders showed significantly increased IL-10 production at week 12 or 24. (*p<0,05; **p<0,01).</p

Th2 cytokine production by PMA/Ionomycin stimulated peripheral blood mononuclear cells.

<p>Prior to therapy, IL-4 and IL-10 production was significantly lower in patients who had subsequent rapid viral decline after 4 weeks of treatment compared to non-SVR group. Baseline Th2 cytokine production did not differ between complete early responders and null-responders (Fig. 4a,b). SVR patients associated with significantly lower baseline IL-10 production compared to non-SVR patients (Fig. 4d).</p

The effect of PEG-IFN plus ribavirin treatment on TNF-α and IL-6 production by TLR-4 stimulated monocytes.

<p>After 12 weeks of PEG-IFN plus RBV treatment, the proinflammatory cytokine production of TLR-4 stimulated monocytes was significantly increased in complete early virological responders compared to null-responder patients. Furthermore, proinflammatory cytokine levels showed no changes and remained low in null-responders throughout antiviral therapy. In contrast to cEVR in RVR patients, proinflammatory cytokine production by monocytes was significantly decreased after 4 weeks of treatment (*p<0,05; **p<0,01 compared to baseline values).</p

Pretreatment proinflammatory cytokine production by Toll-like receptor 4 stimulated monocytes.

<p>Prior to antiviral treatment, TLR-4 agonist induced TNF-α and IL-6 production by peripheral blood monocytes was significantly higher in later rapid virological responder CHC patients (RVR n = 14) compared to complete early virological responders (cEVR n = 19), null-responders (NR n = 17) or healthy controls (HC n = 20) (Fig. 1a,b). Baseline TLR-4 agonist induced proinflammatory cytokine production was similar in cEVR and NR groups. Sustained virological responders (SVR) had significantly higher baseline TNF-α production compared to patients without SVR (non-SVR).</p

Patients’ baseline characteristics.

<p>Baseline HCV RNA levels were significantly lower in rapid virological responders (RVR) compared to complete early virological responders (cEVR) and null-responders (NR). Pretreatment histology and ALT did not differ significantly between study groups. Results are expressed as mean±SE (*p<0,05; **p<0,01).</p><p>(SVR = sustained virological response, BMI = body mass index, HAI = Knodell histological activity index, ALT = alanine amino transferase).</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

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

Correlation between the CTM and the ART in samples with quantifiable HCV RNA levels.

<p>r: Spearman correlation; p: p-value.</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