Fig 1.
#: Samples were drawn at end of treatment (EOT) for most of the cases other than two cases that suffered from viral breakthrough (VBT). Abbreviations: IFN, interferon; DAAs, direct-acting anti-viral agents; D/A; daclatasvir and asuneprevir; S/L: sofosbuvir and ledipasvir; HCC, hepatocellular carcinoma; Pre, pre-treatment; EOT, end of treatment, PBMC, peripheral blood mononuclear cells; SMV, simeprevir; PEG, pegylated, RBV, ribavirin;.
Table 1.
Clinical characteristics of IFN−/DAA group, and comparison of cases stratified by early-emerging HCC.
Table 2.
Clinical characteristics of IFN−/DAA-FU group, and comparison of cases stratified by early-emerging HCC.
Table 3.
Models of multivariate analysis in IFN-/DAA group predicting early- emerging HCC.
Fig 2.
Pre-treatment NKG2D expression and frequencies of natural killer cell compartments stratified by early emergence of HCC or not (IFN-/DAA group).
Filled circles represent cases with early emerging HCC and open circles represent cases without. Statistics were shown as mean with SD. *, P< 0.05; **, P< 0.01.
Fig 3.
Background pre-treatment characteristics and NKG2D expression.
Pre-treatment NKG2D expression was, (a) compared by sex, SVR status, FIB-4 score, and HCC past history with case numbers shown below each group; (b) compared by early emergence of HCC or not in cases restricted to advanced fibrosis (FIB-4 score over 3.25; left panel) and cirrhosis (APRI over 2.0; right panel); (c) compared between cases with or without HCC past history as cases restricted to advanced fibrosis (FIB-4 score over 3.25). Filled circles represent cases with early-emerging HCC and open circles represent cases without. Statistics were shown as mean with SD. *, P< 0.05; NS, not significant.
Table 4.
Various pre-treatment factors and their correlation to pre-treatment NKG2D expression.
Fig 4.
Pre- and End of Treatment NKG2D Expression of NK cells in IFN−/DAA-FU or IFN+ group.
(a) Pre and EOT NKG2D expression of NK cells in each cases of IFN−/DAA-FU group. # represents cases with early-emerging HCC. (b, c) NKG2D Expression of NK cells in IFN−/DAA-FU group and in IFN+ group were compared between cases with or without early emergence of clinically evident HCC. Filled circles (pre) or triangles (EOT), cases with HCC; open circles (pre) or triangles (EOT), cases without HCC. Statistics were shown as median with interquartile ranges. *, P< 0.05; **, P< 0.01; NS, not significant.
Fig 5.
CD25high helper T cells and the correlation to NKG2D expression of NK cells.
(a) CD25high helper T cells in IFN−/DAA-FU group and IFN+ group were compared between cases with or without early emerging HCC. (b) Correlations between pre-treatment NKG2D expression of NK cells and the compartment of CD25high helper T cells in IFN−/DAA group. (c) Correlations between pre- and end of treatment changes in NKG2D expressions (ΔNKG2D) of NK cells and the changes in the compartment of CD25high helper T cells (ΔCD25high/CD4+), in IFN−/DAA-FU group. Filled circles (pre) or triangles (EOT), cases with HCC; open circles (pre) or triangles (EOT), cases without HCC. Statistics were shown as median with interquartile ranges. *, P< 0.05; **, P< 0.01.
Fig 6.
On-treatment decreased of NKG2D correlated to early-emerging HCC.
(a) Changes of NKG2D expression from pre to EOT (ΔNKG2D) were compared in the IFN−/DAA-FU group. The ROC analysis of ΔNKG2D and AFP at EOT as a predictive factor to early HCC emergence were shown. (B) Correlations of ΔNKG2D to other background characteristics and biochemical factors in the IFN−/DAA-FU group were shown. Y-axis: ΔNKG2D expression on NK cells. Filled circles (pre) or triangles (EOT), cases with HCC; open circles (pre) or triangles (EOT), cases without HCC. Statistics were shown as median with interquartile ranges. *, P< 0.05; **, P< 0.01.