Reviewer's report
Title:High mobility group B1 impairs hepatocyte regeneration in acetaminophen
hepatotoxicity
Version: 2Date:6 February 2012
Reviewer: Leo A van Grunsven
Reviewer's report:
High mobility group B1 impairs hepatocyte regeneration in acetaminophen
hepatotoxicity
Runkuan Yang, Antonella Cotoia, Jyrki Tenhunen
- Major Compulsory Revisions
The role of HMGB1 and its potential as a biomarker of prognosis in
acetaminophen hepatotoxicity is a much investigated topic. Yang et al. show in
the APAP mouse model that antibodies against anti-HMGB1 reduce the number
of hepatic inflammatory cells and restore liver damage. Some efforts have been
done to give some mechanistic insight, such as NF-kB EMSAs and cyclin D1
protein level detections. However, while the blocking effect of anti-HMGB1 is very
interesting this has been shown already in previous studies published in 2010 by
the group of Park (Antoine, D. J., et al. (2010) Mol Med 16, 479-490).
A small citation from this paper:
“Neutralizing Circulating HMGB1 Prevents Inflammatory Cell Recruitment after
APAP Treatment We investigated whether modulation of the level of the reduced
form of HMGB1 had any effect on the presence and degree of hepatic
inflammatory cell recruitment in fasted mice 24 h after treatment. There was
significant reduction in the levels of serum hepatic necrosis markers (ALT activity
and full-length K18) in the APAP + anti-HMGB1 antibody group compared with
the APAP + control IgY group (Figure 7B, C). Treatment of animals with a
neutralizing antibody to HMGB1 resulted in increased survival after APAP
challenge compared with that observed in IgY-treated animals (Figure 7A). The
histological findings were in accordance with these results. Although centrilobular
cell loss was seen in the livers of both groups, this was more pronounced in the
APAP + control IgY group (average score 2.7) compared with the APAP +
anti-HMGB1 antibody group (average score 2.2) (Figure 6E, F).”
It would indeed be good to investigate the mechanism but this has not been
thoroughly addressed in this paper. Moreover, the Antoine et al. paper has not
even been cited.
- Minor Essential Revisions
Figure 1.
It is not clear what each band stands for. N=5 for each group is mentioned, does
this mean for each band. This would equal 20 mice in total for this experiment,
this does not correlate with the material and methods.
Figure 2
This piece of text is written in a confusing manner. They state that “compared to
sham IgG treatment, blockade of HMGB1 significantly reduced serum
concentrations of ALT and AST”. However, this is only true for 48 hours after
APAP injection and there is no difference at 24 hours. The authors seem to state
this but again confusing “Anti-HMGB1 therapy did not statistically decrease
serum ALT/AST than that in the sham IgG group (n=6 for each group)”. If a time
is given in this sentence it would make more sense.
Figure 3.
A typical picture is shown.. Low magnification images should be shown.
Immages of stainings of inflammatory cells would be nice as well. Leaving out a
graph for the MPO levels is good, but since there was no real significant
decrease, one does not need to describe this in almost 6 sentences.
Figure 4.
As the authors already mention in their results section “NF-#B is a pleiotropic
transcription factor”, showing that in total liver extracts there is increased NF-#B
binding in an EMSA does not really mean much. In addition, no supershifts are
done nor competitions with mutants or cold probes. One cannot really draw
conclusions from these observations
Figure 5.
Using cyclin D1 westerns as only marker for proliferation is not saying much. An
immunostaining would be good to confirm their observations. Also BrdU or just
Ki67 stainings can also be carried out.
Level of interest: An article of limited interest
Quality of written English: Acceptable
Statistical review: No, the manuscript does not need to be seen by a
statistician.
Declaration of competing interests:
I declare that I have no competing interests
Reviewer's report
Title:High mobility group B1 impairs hepatocyte regeneration in acetaminophen
hepatotoxicity
Version: 2Date:6 February 2012
Reviewer: Leo A van Grunsven
Reviewer's report:
High mobility group B1 impairs hepatocyte regeneration in acetaminophen
hepatotoxicity
Runkuan Yang, Antonella Cotoia, Jyrki Tenhunen
- Major Compulsory Revisions
The role of HMGB1 and its potential as a biomarker of prognosis in
acetaminophen hepatotoxicity is a much investigated topic. Yang et al. show in
the APAP mouse model that antibodies against anti-HMGB1 reduce the number
of hepatic inflammatory cells and restore liver damage. Some efforts have been
done to give some mechanistic insight, such as NF-kB EMSAs and cyclin D1
protein level detections. However, while the blocking effect of anti-HMGB1 is very
interesting this has been shown already in previous studies published in 2010 by
the group of Park (Antoine, D. J., et al. (2010) Mol Med 16, 479-490).
A small citation from this paper:
“Neutralizing Circulating HMGB1 Prevents Inflammatory Cell Recruitment after
APAP Treatment We investigated whether modulation of the level of the reduced
form of HMGB1 had any effect on the presence and degree of hepatic
inflammatory cell recruitment in fasted mice 24 h after treatment. There was
significant reduction in the levels of serum hepatic necrosis markers (ALT activity
and full-length K18) in the APAP + anti-HMGB1 antibody group compared with
the APAP + control IgY group (Figure 7B, C). Treatment of animals with a
neutralizing antibody to HMGB1 resulted in increased survival after APAP
challenge compared with that observed in IgY-treated animals (Figure 7A). The
histological findings were in accordance with these results. Although centrilobular
cell loss was seen in the livers of both groups, this was more pronounced in the
APAP + control IgY group (average score 2.7) compared with the APAP +
anti-HMGB1 antibody group (average score 2.2) (Figure 6E, F).”
It would indeed be good to investigate the mechanism but this has not been
thoroughly addressed in this paper. Moreover, the Antoine et al. paper has not
even been cited.
- Minor Essential Revisions
Figure 1.
It is not clear what each band stands for. N=5 for each group is mentioned, does
this mean for each band. This would equal 20 mice in total for this experiment,
this does not correlate with the material and methods.
Figure 2
This piece of text is written in a confusing manner. They state that “compared to
sham IgG treatment, blockade of HMGB1 significantly reduced serum
concentrations of ALT and AST”. However, this is only true for 48 hours after
APAP injection and there is no difference at 24 hours. The authors seem to state
this but again confusing “Anti-HMGB1 therapy did not statistically decrease
serum ALT/AST than that in the sham IgG group (n=6 for each group)”. If a time
is given in this sentence it would make more sense.
Figure 3.
A typical picture is shown.. Low magnification images should be shown.
Immages of stainings of inflammatory cells would be nice as well. Leaving out a
graph for the MPO levels is good, but since there was no real significant
decrease, one does not need to describe this in almost 6 sentences.
Figure 4.
As the authors already mention in their results section “NF-#B is a pleiotropic
transcription factor”, showing that in total liver extracts there is increased NF-#B
binding in an EMSA does not really mean much. In addition, no supershifts are
done nor competitions with mutants or cold probes. One cannot really draw
conclusions from these observations
Figure 5.
Using cyclin D1 westerns as only marker for proliferation is not saying much. An
immunostaining would be good to confirm their observations. Also BrdU or just
Ki67 stainings can also be carried out.
Level of interest: An article of limited interest
Quality of written English: Acceptable
Statistical review: No, the manuscript does not need to be seen by a
statistician.
Declaration of competing interests:
I declare that I have no competing interests
Please review this paper.
user
Reviewer's report
Title:High mobility group B1 impairs hepatocyte regeneration in acetaminophen
hepatotoxicity
Version: 2Date:6 February 2012
Reviewer: Leo A van Grunsven
Reviewer's report:
High mobility group B1 impairs hepatocyte regeneration in acetaminophen
hepatotoxicity
Runkuan Yang, Antonella Cotoia, Jyrki Tenhunen
- Major Compulsory Revisions
The role of HMGB1 and its potential as a biomarker of prognosis in
acetaminophen hepatotoxicity is a much investigated topic. Yang et al. show in
the APAP mouse model that antibodies against anti-HMGB1 reduce the number
of hepatic inflammatory cells and restore liver damage. Some efforts have been
done to give some mechanistic insight, such as NF-kB EMSAs and cyclin D1
protein level detections. However, while the blocking effect of anti-HMGB1 is very
interesting this has been shown already in previous studies published in 2010 by
the group of Park (Antoine, D. J., et al. (2010) Mol Med 16, 479-490).
A small citation from this paper:
“Neutralizing Circulating HMGB1 Prevents Inflammatory Cell Recruitment after
APAP Treatment We investigated whether modulation of the level of the reduced
form of HMGB1 had any effect on the presence and degree of hepatic
inflammatory cell recruitment in fasted mice 24 h after treatment. There was
significant reduction in the levels of serum hepatic necrosis markers (ALT activity
and full-length K18) in the APAP + anti-HMGB1 antibody group compared with
the APAP + control IgY group (Figure 7B, C). Treatment of animals with a
neutralizing antibody to HMGB1 resulted in increased survival after APAP
challenge compared with that observed in IgY-treated animals (Figure 7A). The
histological findings were in accordance with these results. Although centrilobular
cell loss was seen in the livers of both groups, this was more pronounced in the
APAP + control IgY group (average score 2.7) compared with the APAP +
anti-HMGB1 antibody group (average score 2.2) (Figure 6E, F).”
It would indeed be good to investigate the mechanism but this has not been
thoroughly addressed in this paper. Moreover, the Antoine et al. paper has not
even been cited.
- Minor Essential Revisions
Figure 1.
It is not clear what each band stands for. N=5 for each group is mentioned, does
this mean for each band. This would equal 20 mice in total for this experiment,
this does not correlate with the material and methods.
Figure 2
This piece of text is written in a confusing manner. They state that “compared to
sham IgG treatment, blockade of HMGB1 significantly reduced serum
concentrations of ALT and AST”. However, this is only true for 48 hours after
APAP injection and there is no difference at 24 hours. The authors seem to state
this but again confusing “Anti-HMGB1 therapy did not statistically decrease
serum ALT/AST than that in the sham IgG group (n=6 for each group)”. If a time
is given in this sentence it would make more sense.
Figure 3.
A typical picture is shown.. Low magnification images should be shown.
Immages of stainings of inflammatory cells would be nice as well. Leaving out a
graph for the MPO levels is good, but since there was no real significant
decrease, one does not need to describe this in almost 6 sentences.
Figure 4.
As the authors already mention in their results section “NF-#B is a pleiotropic
transcription factor”, showing that in total liver extracts there is increased NF-#B
binding in an EMSA does not really mean much. In addition, no supershifts are
done nor competitions with mutants or cold probes. One cannot really draw
conclusions from these observations
Figure 5.
Using cyclin D1 westerns as only marker for proliferation is not saying much. An
immunostaining would be good to confirm their observations. Also BrdU or just
Ki67 stainings can also be carried out.
Level of interest: An article of limited interest
Quality of written English: Acceptable
Statistical review: No, the manuscript does not need to be seen by a
statistician.
Declaration of competing interests:
I declare that I have no competing interests
assistant