Title: Natural killer cell activity is a risk factor for the recurrence risk after curative treatment
of hepatocellular carcinoma
Version: 1 Date: 17 Apr 2021
Reviewer's report:
In this study, Lee et al. investigated the association between NK cell activity and the recurrence
risk of HCC. They examined IFN-g production from peripheral blood NK cells at the time of
diagnosis and 1 month after treatment. Importantly, they found that patients with low NK cell
activity (<45% IFN-g producing NK cells) at 1 month after treatment showed a significantly
higher HCC recurrence rate than patients with high NK cell activity. They concluded that
decreased NK cell activity is associated with the increased recurrence risk of HCC after
treatment. I have several technical comments.
1. The authors need to present representative FACS plots for IFN-g production from NK cells.
It will be better to show examples of high and low NK cell activities, respectively.
2. In this study, PBMCs were stimulated with Leukocyte Activation Cocktail (BD Biosciences)
to induce IFN-g production from NK cells. Leukocyte Activation Cocktail (BD Biosciences)
includes PMA and ionomycin and can stimulate not only NK cells but also T cells. It means that
the authors can analyze IFN-g production from T cells without further experiments. They can
re-analyze their FACS data files by gating CD3+ T cells. Please analyze a possible association
of T cell activity with stage and recurrence risk of HCC. This analysis will validate whether the
conclusion of this study is NK cell-specific or not.
3. In supplementary Figure 2, representative FACS plots for cases with preserved (A) or
decreased (B) NK cell proportion are presented. However, it seems that there is no difference in
NK cell proportion between the two cases. It seems that cell viability is poor in the second case
(B). What was cell viability of PBMC in general? Regarding cell viability, my above comment
(#2) is important because poor cell viability results in poor IFN-g production not only from NK
cells but also from T cells.
Title: Natural killer cell activity is a risk factor for the recurrence risk after curative treatment
of hepatocellular carcinoma
Version: 1 Date: 17 Apr 2021
Reviewer's report:
In this study, Lee et al. investigated the association between NK cell activity and the recurrence
risk of HCC. They examined IFN-g production from peripheral blood NK cells at the time of
diagnosis and 1 month after treatment. Importantly, they found that patients with low NK cell
activity (<45% IFN-g producing NK cells) at 1 month after treatment showed a significantly
higher HCC recurrence rate than patients with high NK cell activity. They concluded that
decreased NK cell activity is associated with the increased recurrence risk of HCC after
treatment. I have several technical comments.
1. The authors need to present representative FACS plots for IFN-g production from NK cells.
It will be better to show examples of high and low NK cell activities, respectively.
2. In this study, PBMCs were stimulated with Leukocyte Activation Cocktail (BD Biosciences)
to induce IFN-g production from NK cells. Leukocyte Activation Cocktail (BD Biosciences)
includes PMA and ionomycin and can stimulate not only NK cells but also T cells. It means that
the authors can analyze IFN-g production from T cells without further experiments. They can
re-analyze their FACS data files by gating CD3+ T cells. Please analyze a possible association
of T cell activity with stage and recurrence risk of HCC. This analysis will validate whether the
conclusion of this study is NK cell-specific or not.
3. In supplementary Figure 2, representative FACS plots for cases with preserved (A) or
decreased (B) NK cell proportion are presented. However, it seems that there is no difference in
NK cell proportion between the two cases. It seems that cell viability is poor in the second case
(B). What was cell viability of PBMC in general? Regarding cell viability, my above comment
(#2) is important because poor cell viability results in poor IFN-g production not only from NK
cells but also from T cells.
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: 17 Apr 2021
Reviewer's report:
In this study, Lee et al. investigated the association between NK cell activity and the recurrence
risk of HCC. They examined IFN-g production from peripheral blood NK cells at the time of
diagnosis and 1 month after treatment. Importantly, they found that patients with low NK cell
activity (<45% IFN-g producing NK cells) at 1 month after treatment showed a significantly
higher HCC recurrence rate than patients with high NK cell activity. They concluded that
decreased NK cell activity is associated with the increased recurrence risk of HCC after
treatment. I have several technical comments.
1. The authors need to present representative FACS plots for IFN-g production from NK cells.
It will be better to show examples of high and low NK cell activities, respectively.
2. In this study, PBMCs were stimulated with Leukocyte Activation Cocktail (BD Biosciences)
to induce IFN-g production from NK cells. Leukocyte Activation Cocktail (BD Biosciences)
includes PMA and ionomycin and can stimulate not only NK cells but also T cells. It means that
the authors can analyze IFN-g production from T cells without further experiments. They can
re-analyze their FACS data files by gating CD3+ T cells. Please analyze a possible association
of T cell activity with stage and recurrence risk of HCC. This analysis will validate whether the
conclusion of this study is NK cell-specific or not.
3. In supplementary Figure 2, representative FACS plots for cases with preserved (A) or
decreased (B) NK cell proportion are presented. However, it seems that there is no difference in
NK cell proportion between the two cases. It seems that cell viability is poor in the second case
(B). What was cell viability of PBMC in general? Regarding cell viability, my above comment
(#2) is important because poor cell viability results in poor IFN-g production not only from NK
cells but also from T cells.
assistant