Title: Natural killer cell activity is a risk factor for the recurrence risk after curative treatment
of hepatocellular carcinoma
Version: 1 Date: 13 Apr 2021
Reviewer's report:
This study aimed to investigate the role of natural killer (NK) cell activity in determining
the outcomes of patients with HCC. The authors demonstrated that the IFN-# producing NK
cell proportion seemed to be lower in patients with advanced stage HCC when compared to
those with early stage HCC. Moreover, for patients who underwent curative therapy, the IFN-#
producing NK cell proportion < 45% at 1 month after treatment was associated with an increased
risk of recurrence after therapy.
Immunopathogenesis of HCC has not been clearly elucidated, and this article is very timely in
that it explains, at least partly, the important role of NK cells (the most important innate immune
cells for the cancer control) in HCC control.
1. The authors seem to have used pan-T/NK cell stimulants for the NK cell stimulation and ICCS
for IFN-gamma, because the authors divided the population into patients with IFN-g+NK >45%
or <45%.  Representative FACS dotplots for IFN-gamma staining (with appropriate gating) of
NK cells with isotype control should be presented as a main figure.
2. In chronic HBV infection, NK cells have reported to show cytotoxicity against HBV-
specific CD8 T cells to weaken adaptive immune responses. Moreover, it was also reported
that peripheral NK-cell activity (cytokine secretion and cytotoxicity) in immune active HBV
infection has a negative correlation with  (Li et al. PLoS One. 2014;9:e86927). As a majority
of included patients had chronic hepatitis B as an underlying liver disease in this study, patients
should be stratified according to the serum HBV DNA levels.
3. It would be better to define primary outcome more precisely: Was it a recurrence-free survival
or time-to-progression? It might be an important issue since patients with advanced or end-stage
HCC were also included.
4.  In this study, there were no significant changes in IFN-# producing NK cell proportion from
baseline to 1 month after curative treatment. However, only the post-treatment, but not baseline
IFN-# producing NK cell proportion < 45% was associated with tumor recurrence. It is suggested
to provide the Figures to depict the correlation between baseline and post-treatment NK cell
activity.
5. Minor spelling and spacing errors are noted throughout the manuscript. Moreover, awkward
expression are noted. One more round of English editing should be performed.
Title: Natural killer cell activity is a risk factor for the recurrence risk after curative treatment
of hepatocellular carcinoma
Version: 1 Date: 13 Apr 2021
Reviewer's report:
This study aimed to investigate the role of natural killer (NK) cell activity in determining
the outcomes of patients with HCC. The authors demonstrated that the IFN-# producing NK
cell proportion seemed to be lower in patients with advanced stage HCC when compared to
those with early stage HCC. Moreover, for patients who underwent curative therapy, the IFN-#
producing NK cell proportion < 45% at 1 month after treatment was associated with an increased
risk of recurrence after therapy.
Immunopathogenesis of HCC has not been clearly elucidated, and this article is very timely in
that it explains, at least partly, the important role of NK cells (the most important innate immune
cells for the cancer control) in HCC control.
1. The authors seem to have used pan-T/NK cell stimulants for the NK cell stimulation and ICCS
for IFN-gamma, because the authors divided the population into patients with IFN-g+NK >45%
or <45%.  Representative FACS dotplots for IFN-gamma staining (with appropriate gating) of
NK cells with isotype control should be presented as a main figure.
2. In chronic HBV infection, NK cells have reported to show cytotoxicity against HBV-
specific CD8 T cells to weaken adaptive immune responses. Moreover, it was also reported
that peripheral NK-cell activity (cytokine secretion and cytotoxicity) in immune active HBV
infection has a negative correlation with  (Li et al. PLoS One. 2014;9:e86927). As a majority
of included patients had chronic hepatitis B as an underlying liver disease in this study, patients
should be stratified according to the serum HBV DNA levels.
3. It would be better to define primary outcome more precisely: Was it a recurrence-free survival
or time-to-progression? It might be an important issue since patients with advanced or end-stage
HCC were also included.
4.  In this study, there were no significant changes in IFN-# producing NK cell proportion from
baseline to 1 month after curative treatment. However, only the post-treatment, but not baseline
IFN-# producing NK cell proportion < 45% was associated with tumor recurrence. It is suggested
to provide the Figures to depict the correlation between baseline and post-treatment NK cell
activity.
5. Minor spelling and spacing errors are noted throughout the manuscript. Moreover, awkward
expression are noted. One more round of English editing should be performed.
Please review this paper.
user
Title: Natural killer cell activity is a risk factor for the recurrence risk after curative treatment
of hepatocellular carcinoma
Version: 1 Date: 13 Apr 2021
Reviewer's report:
This study aimed to investigate the role of natural killer (NK) cell activity in determining
the outcomes of patients with HCC. The authors demonstrated that the IFN-# producing NK
cell proportion seemed to be lower in patients with advanced stage HCC when compared to
those with early stage HCC. Moreover, for patients who underwent curative therapy, the IFN-#
producing NK cell proportion < 45% at 1 month after treatment was associated with an increased
risk of recurrence after therapy.
Immunopathogenesis of HCC has not been clearly elucidated, and this article is very timely in
that it explains, at least partly, the important role of NK cells (the most important innate immune
cells for the cancer control) in HCC control.
1. The authors seem to have used pan-T/NK cell stimulants for the NK cell stimulation and ICCS
for IFN-gamma, because the authors divided the population into patients with IFN-g+NK >45%
or <45%.  Representative FACS dotplots for IFN-gamma staining (with appropriate gating) of
NK cells with isotype control should be presented as a main figure.
2. In chronic HBV infection, NK cells have reported to show cytotoxicity against HBV-
specific CD8 T cells to weaken adaptive immune responses. Moreover, it was also reported
that peripheral NK-cell activity (cytokine secretion and cytotoxicity) in immune active HBV
infection has a negative correlation with  (Li et al. PLoS One. 2014;9:e86927). As a majority
of included patients had chronic hepatitis B as an underlying liver disease in this study, patients
should be stratified according to the serum HBV DNA levels.
3. It would be better to define primary outcome more precisely: Was it a recurrence-free survival
or time-to-progression? It might be an important issue since patients with advanced or end-stage
HCC were also included.
4.  In this study, there were no significant changes in IFN-# producing NK cell proportion from
baseline to 1 month after curative treatment. However, only the post-treatment, but not baseline
IFN-# producing NK cell proportion < 45% was associated with tumor recurrence. It is suggested
to provide the Figures to depict the correlation between baseline and post-treatment NK cell
activity.
5. Minor spelling and spacing errors are noted throughout the manuscript. Moreover, awkward
expression are noted. One more round of English editing should be performed.
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