Reviewer ’s report  
 
Title: Approaching treatment for immunological rejection of living -donor liver transplantation in 
rats 
 
Version: 3 Date: 23 Jul 2019  
 
Reviewer: Reviewer 2  
 
Reviewer's report:  
 
Editor Comments:  
 
A point -by-point response letter must accompany your re vised manuscript. This letter must 
provide a detailed response to each reviewer/editorial point raised, describing exactly what 
amendments have been made to the manuscript text and where these can be viewed (e.g. 
Methods section, line 12, page 5). If you d isagree with any comments raised, please provide a 
detailed rebuttal to help explain and justify your decision.  
 
At this stage, we ask that you submit a clean version of your manuscript and do not include track 
changes or highlighting.  
 
 
 
Reviewer reports:  
 
Lucas Souto Nacif, PhD (Reviewer 1): Acceptable corrections and manuscript imporvements  
Thank the reviewer for your suggestions.  
 
Reviewer 2 (Reviewer 2): GENERAL COMMENTS: The authors replied to all the reviewers' 
comments and requests.  
 
 
 
REQUESTED REVISI ONS:  
 
However, some requests have not been fulfilled  
 
1 display of isotype control in Fig.4;  
 
Thank the reviewer for your suggestion, we listed the isotype control in the last responsiveness 
letter as below, since most of paper did not use the isotype cont rol in final figure, and it will not 
be well rearranged, I can list isotype control as supplement data if you agree with it.  
 
 
 
2 partial clarification on the limitations of the study in the Conclusion session).  
Thank the reviewer for your suggestion, we add the limitation in Conclusion session, see line 
389 
 
3 Limitation in analysis of hepatic metabolic enzymes, such as Cyp2C or Cyp3A1 raised by 
reviewer #2, has not been stated while should be properly addressed in Conclusion session and 
clearly stated if object of ongoing studies.   
Thank the reviewer for your suggestion. Although we select CYP3A2 as enzyme involved in 
tacrolimus metabolism was based on a couple of manuscripts (refs 29 and 30), the most abundant 
CYP subfamily (in rat liver) is the CYP2C, which has a role closer to human CYP3A enzymes, 
we list that as this study’s limitation, and will further check it in ongoing study, see line 393  
 
4 The authors agree with reviewer's suggestion to re -arrange Resul ts section. However Figures 
need to be rearrange as well (i.e., lung/kidney histological analysis should be moved in a separate 
Figure or described earlier in the text).  
Thank the reviewer for your suggestion, lung/kidney histological analysis is describe d earlier in 
the text, see line 217  
 
5 Similar comment for figure 3 (apoptosis is described at page 12 while alphabetaTCR at page 
10, but both represented in the same Figure).  
Thank you for your suggestion, alphabetaTCR and apoptosis is described at same page, see line 
231 
 
6 Figure Legends contain large amount of result description. Such sentences should be moved in 
Results, limiting the long text in Legends. Once again, several Figure contains one panel only, 
resulting in large number of figures (8). Sev eral Figures can be rearranged and grouped. Several 
mistypos are still present in the text (i.e. kedney instead of kidney in Figure 2)  
Thank the review for your suggestions, we shortened the figure legends. We also combine the 
figure 5 and figure 7, and c orrect the mistypos.  
 
 
 
ADDITIONAL REQUESTS/SUGGESTIONS:  
 
A final check of the Discussion section may be appropriate. Attempt in shorten it up is highly 
recommended  
 
We try to shorten the discussion section  
 
 
 
 
 
 
 
 
Are the methods appropriate and well described?  
If not, please specify wha t is required in your comments to the authors.  
 
Yes 
 
Does the work include the necessary controls?  
If not, please specify which controls are required in your comments to the authors.  
 
Yes 
 
Are the conclusions drawn adequately supported by the data shown?  
If not , please explain in your comments to the authors.  
 
Yes 
 
Are you able to assess any statistics in the manus cript or would you recommend an 
additional statistical review?  
If an additional statistical review is recommended, please specify what aspects require fu rther 
assessment in your comments to the editors.  
 
I am able to assess the statistics  
 
Quality of written English  
Please indicate the quality of language in the manuscript:  
 
Acceptable Declaration of competing interests  
Please complete a declaration of competing interests, considering the following questions:  
 
1. Have you in the past five years received reimbursements, f ees, funding, or salary from an 
organisation that may in any way gain or lose financially from the publi cation of this 
manuscript, either now or in the future?  
 
2. Do you hold any stocks or shares in an organisation t hat may in any way gain or lose 
financially from the publication of this manuscript, either now or in the future?  
 
3. Do you hold or are you curr ently applying for any patents  relating to the content of the 
manuscript?  
 
4. Have you received reimbursements, fees, funding, or salary from an organization that 
holds or has applied for patents relating to the content of the manuscript?  
 
5. Do you have any  other financial competing interests?  
 
6. Do you have any non -financial competing interests in relation to this paper?  
 
If you can answer no to all of the above, write 'I declare that I have  no competing interests' 
below. If your reply is yes to any, please give details below.  
 
This reviewer has been recruited by a partner organization, Research Square. Reviewers with 
declared or apparent competing interests are not utilized for these reviews. This reviewer has 
agreed to publication of their comments online under a Creative Commons Attribution License 
attributed to Research Square and was paid a small honorarium for compl eting the review within 
a specified timeframe. Honoraria for reviews such as this are paid regardless of the reviewer 
recommendation.  
 
I agree to the open peer review policy of the journal. I understan d that my name will be included 
on my report to the authors and, if the manuscript is accepted f or publication, my named report 
including any att achments I upload will be posted on the website along with the authors' 
responses. I agree for my report to be made available under an Open Access Creative Commons  
CC-BY license ( http://creativecommons.org/licenses/by/4.0/ ). I understand that any comments 
which I do not wish to be included in my named report can be in cluded as confidential comments 
to the editors, which will not be published.  
 
I agree to the open peer review policy of the journal  
Reviewer ’s report  
 
Title: Approaching treatment for immunological rejection of living -donor liver transplantation in 
rats 
 
Version: 3 Date: 23 Jul 2019  
 
Reviewer: Reviewer 2  
 
Reviewer's report:  
 
Editor Comments:  
 
A point -by-point response letter must accompany your re vised manuscript. This letter must 
provide a detailed response to each reviewer/editorial point raised, describing exactly what 
amendments have been made to the manuscript text and where these can be viewed (e.g. 
Methods section, line 12, page 5). If you d isagree with any comments raised, please provide a 
detailed rebuttal to help explain and justify your decision.  
 
At this stage, we ask that you submit a clean version of your manuscript and do not include track 
changes or highlighting.  
 
 
 
Reviewer reports:  
 
Lucas Souto Nacif, PhD (Reviewer 1): Acceptable corrections and manuscript imporvements  
Thank the reviewer for your suggestions.  
 
Reviewer 2 (Reviewer 2): GENERAL COMMENTS: The authors replied to all the reviewers' 
comments and requests.  
 
 
 
REQUESTED REVISI ONS:  
 
However, some requests have not been fulfilled  
 
1 display of isotype control in Fig.4;  
 
Thank the reviewer for your suggestion, we listed the isotype control in the last responsiveness 
letter as below, since most of paper did not use the isotype cont rol in final figure, and it will not 
be well rearranged, I can list isotype control as supplement data if you agree with it.  
 
 
 
2 partial clarification on the limitations of the study in the Conclusion session).  
Thank the reviewer for your suggestion, we add the limitation in Conclusion session, see line 
389 
 
3 Limitation in analysis of hepatic metabolic enzymes, such as Cyp2C or Cyp3A1 raised by 
reviewer #2, has not been stated while should be properly addressed in Conclusion session and 
clearly stated if object of ongoing studies.   
Thank the reviewer for your suggestion. Although we select CYP3A2 as enzyme involved in 
tacrolimus metabolism was based on a couple of manuscripts (refs 29 and 30), the most abundant 
CYP subfamily (in rat liver) is the CYP2C, which has a role closer to human CYP3A enzymes, 
we list that as this study’s limitation, and will further check it in ongoing study, see line 393  
 
4 The authors agree with reviewer's suggestion to re -arrange Resul ts section. However Figures 
need to be rearrange as well (i.e., lung/kidney histological analysis should be moved in a separate 
Figure or described earlier in the text).  
Thank the reviewer for your suggestion, lung/kidney histological analysis is describe d earlier in 
the text, see line 217  
 
5 Similar comment for figure 3 (apoptosis is described at page 12 while alphabetaTCR at page 
10, but both represented in the same Figure).  
Thank you for your suggestion, alphabetaTCR and apoptosis is described at same page, see line 
231 
 
6 Figure Legends contain large amount of result description. Such sentences should be moved in 
Results, limiting the long text in Legends. Once again, several Figure contains one panel only, 
resulting in large number of figures (8). Sev eral Figures can be rearranged and grouped. Several 
mistypos are still present in the text (i.e. kedney instead of kidney in Figure 2)  
Thank the review for your suggestions, we shortened the figure legends. We also combine the 
figure 5 and figure 7, and c orrect the mistypos.  
 
 
 
ADDITIONAL REQUESTS/SUGGESTIONS:  
 
A final check of the Discussion section may be appropriate. Attempt in shorten it up is highly 
recommended  
 
We try to shorten the discussion section  
 
 
 
 
 
 
 
 
Are the methods appropriate and well described?  
If not, please specify wha t is required in your comments to the authors.  
 
Yes 
 
Does the work include the necessary controls?  
If not, please specify which controls are required in your comments to the authors.  
 
Yes 
 
Are the conclusions drawn adequately supported by the data shown?  
If not , please explain in your comments to the authors.  
 
Yes 
 
Are you able to assess any statistics in the manus cript or would you recommend an 
additional statistical review?  
If an additional statistical review is recommended, please specify what aspects require fu rther 
assessment in your comments to the editors.  
 
I am able to assess the statistics  
 
Quality of written English  
Please indicate the quality of language in the manuscript:  
 
Acceptable Declaration of competing interests  
Please complete a declaration of competing interests, considering the following questions:  
 
1. Have you in the past five years received reimbursements, f ees, funding, or salary from an 
organisation that may in any way gain or lose financially from the publi cation of this 
manuscript, either now or in the future?  
 
2. Do you hold any stocks or shares in an organisation t hat may in any way gain or lose 
financially from the publication of this manuscript, either now or in the future?  
 
3. Do you hold or are you curr ently applying for any patents  relating to the content of the 
manuscript?  
 
4. Have you received reimbursements, fees, funding, or salary from an organization that 
holds or has applied for patents relating to the content of the manuscript?  
 
5. Do you have any  other financial competing interests?  
 
6. Do you have any non -financial competing interests in relation to this paper?  
 
If you can answer no to all of the above, write 'I declare that I have  no competing interests' 
below. If your reply is yes to any, please give details below.  
 
This reviewer has been recruited by a partner organization, Research Square. Reviewers with 
declared or apparent competing interests are not utilized for these reviews. This reviewer has 
agreed to publication of their comments online under a Creative Commons Attribution License 
attributed to Research Square and was paid a small honorarium for compl eting the review within 
a specified timeframe. Honoraria for reviews such as this are paid regardless of the reviewer 
recommendation.  
 
I agree to the open peer review policy of the journal. I understan d that my name will be included 
on my report to the authors and, if the manuscript is accepted f or publication, my named report 
including any att achments I upload will be posted on the website along with the authors' 
responses. I agree for my report to be made available under an Open Access Creative Commons  
CC-BY license ( http://creativecommons.org/licenses/by/4.0/ ). I understand that any comments 
which I do not wish to be included in my named report can be in cluded as confidential comments 
to the editors, which will not be published.  
 
I agree to the open peer review policy of the journal  
Please review this paper.
user
Reviewer ’s report  
 
Title: Approaching treatment for immunological rejection of living -donor liver transplantation in 
rats 
 
Version: 3 Date: 23 Jul 2019  
 
Reviewer: Reviewer 2  
 
Reviewer's report:  
 
Editor Comments:  
 
A point -by-point response letter must accompany your re vised manuscript. This letter must 
provide a detailed response to each reviewer/editorial point raised, describing exactly what 
amendments have been made to the manuscript text and where these can be viewed (e.g. 
Methods section, line 12, page 5). If you d isagree with any comments raised, please provide a 
detailed rebuttal to help explain and justify your decision.  
 
At this stage, we ask that you submit a clean version of your manuscript and do not include track 
changes or highlighting.  
 
 
 
Reviewer reports:  
 
Lucas Souto Nacif, PhD (Reviewer 1): Acceptable corrections and manuscript imporvements  
Thank the reviewer for your suggestions.  
 
Reviewer 2 (Reviewer 2): GENERAL COMMENTS: The authors replied to all the reviewers' 
comments and requests.  
 
 
 
REQUESTED REVISI ONS:  
 
However, some requests have not been fulfilled  
 
1 display of isotype control in Fig.4;  
 
Thank the reviewer for your suggestion, we listed the isotype control in the last responsiveness 
letter as below, since most of paper did not use the isotype cont rol in final figure, and it will not 
be well rearranged, I can list isotype control as supplement data if you agree with it.  
 
 
 
2 partial clarification on the limitations of the study in the Conclusion session).  
Thank the reviewer for your suggestion, we add the limitation in Conclusion session, see line 
389 
 
3 Limitation in analysis of hepatic metabolic enzymes, such as Cyp2C or Cyp3A1 raised by 
reviewer #2, has not been stated while should be properly addressed in Conclusion session and 
clearly stated if object of ongoing studies.   
Thank the reviewer for your suggestion. Although we select CYP3A2 as enzyme involved in 
tacrolimus metabolism was based on a couple of manuscripts (refs 29 and 30), the most abundant 
CYP subfamily (in rat liver) is the CYP2C, which has a role closer to human CYP3A enzymes, 
we list that as this study’s limitation, and will further check it in ongoing study, see line 393  
 
4 The authors agree with reviewer's suggestion to re -arrange Resul ts section. However Figures 
need to be rearrange as well (i.e., lung/kidney histological analysis should be moved in a separate 
Figure or described earlier in the text).  
Thank the reviewer for your suggestion, lung/kidney histological analysis is describe d earlier in 
the text, see line 217  
 
5 Similar comment for figure 3 (apoptosis is described at page 12 while alphabetaTCR at page 
10, but both represented in the same Figure).  
Thank you for your suggestion, alphabetaTCR and apoptosis is described at same page, see line 
231 
 
6 Figure Legends contain large amount of result description. Such sentences should be moved in 
Results, limiting the long text in Legends. Once again, several Figure contains one panel only, 
resulting in large number of figures (8). Sev eral Figures can be rearranged and grouped. Several 
mistypos are still present in the text (i.e. kedney instead of kidney in Figure 2)  
Thank the review for your suggestions, we shortened the figure legends. We also combine the 
figure 5 and figure 7, and c orrect the mistypos.  
 
 
 
ADDITIONAL REQUESTS/SUGGESTIONS:  
 
A final check of the Discussion section may be appropriate. Attempt in shorten it up is highly 
recommended  
 
We try to shorten the discussion section  
 
 
 
 
 
 
 
 
Are the methods appropriate and well described?  
If not, please specify wha t is required in your comments to the authors.  
 
Yes 
 
Does the work include the necessary controls?  
If not, please specify which controls are required in your comments to the authors.  
 
Yes 
 
Are the conclusions drawn adequately supported by the data shown?  
If not , please explain in your comments to the authors.  
 
Yes 
 
Are you able to assess any statistics in the manus cript or would you recommend an 
additional statistical review?  
If an additional statistical review is recommended, please specify what aspects require fu rther 
assessment in your comments to the editors.  
 
I am able to assess the statistics  
 
Quality of written English  
Please indicate the quality of language in the manuscript:  
 
Acceptable Declaration of competing interests  
Please complete a declaration of competing interests, considering the following questions:  
 
1. Have you in the past five years received reimbursements, f ees, funding, or salary from an 
organisation that may in any way gain or lose financially from the publi cation of this 
manuscript, either now or in the future?  
 
2. Do you hold any stocks or shares in an organisation t hat may in any way gain or lose 
financially from the publication of this manuscript, either now or in the future?  
 
3. Do you hold or are you curr ently applying for any patents  relating to the content of the 
manuscript?  
 
4. Have you received reimbursements, fees, funding, or salary from an organization that 
holds or has applied for patents relating to the content of the manuscript?  
 
5. Do you have any  other financial competing interests?  
 
6. Do you have any non -financial competing interests in relation to this paper?  
 
If you can answer no to all of the above, write 'I declare that I have  no competing interests' 
below. If your reply is yes to any, please give details below.  
 
This reviewer has been recruited by a partner organization, Research Square. Reviewers with 
declared or apparent competing interests are not utilized for these reviews. This reviewer has 
agreed to publication of their comments online under a Creative Commons Attribution License 
attributed to Research Square and was paid a small honorarium for compl eting the review within 
a specified timeframe. Honoraria for reviews such as this are paid regardless of the reviewer 
recommendation.  
 
I agree to the open peer review policy of the journal. I understan d that my name will be included 
on my report to the authors and, if the manuscript is accepted f or publication, my named report 
including any att achments I upload will be posted on the website along with the authors' 
responses. I agree for my report to be made available under an Open Access Creative Commons  
CC-BY license ( http://creativecommons.org/licenses/by/4.0/ ). I understand that any comments 
which I do not wish to be included in my named report can be in cluded as confidential comments 
to the editors, which will not be published.  
 
I agree to the open peer review policy of the journal  
assistant