Reviewer ’s report  
 
Title: HBsAg quantification predicts off -treatment response to interferon in chronic hepatitis B  
patients：A retrospective study of 250 cases  
 
Version: 0 Date: 29 Jan 2020  
 
Reviewer: Research Square Reviewer 1  
 
Reviewer's report:  
 
PEER REVIEWER ASSESSMENTS:  
 
OBJECTIV E - Full research articles: is there a clear objective that addresses a testable research 
question(s) (brief or other article types: is there a clear objective)?  
Yes - there is a clear objective  
 
DESIGN - Is the current approach (including controls and ana lysis protocols) appropriate for the 
objective?  
Yes - the approach is appropriate  
 
EXECUTION - Are the experiments and analyses performed with technical rigor to allow 
confidence in the results?  
Yes - experiments and analyses were performed appropriately  
 
STATISTICS - Is the use of statistics in the manuscript appropriate?  
Yes - appropriate statistical analyses have been used in the study  
 
INTERPRETATION - Is the current interpretation/discussion of the results reasonable and not 
overstated?  
Yes - the autho r's interpretation is reasonable  
 
OVERALL MANUSCRIPT POTENTIAL - Is the current version of this work technically 
sound? If not, can revisions be made to make the work technically sound?  
Probably - with minor revisions  
 
 
PEER REVIEWER COMMENTS:  
 
GENERAL COM MENTS:  
 
In this manuscript, the authors report the results of a single -center, retrospective cohort study 
which aimed to examine the predictive value of end -of-treatment quantitative HBsAg titer in 
identifying patients who are likely to achieve off -treatme nt HBsAg loss in response to interferon 
therapy.  Using a cohort of 250 patients who underwent IFN therapy for chronic HBV infection 
between 2010 -2017, EOT HBsAg level &lt;10 IU/mL was associated with 30.4% cumulative 
HBsAg loss rate 5 years post -treatment , compared with 9.8% (HBsAg 10 -200 IU/mL) or 0% 
(HBsAg &gt;200 IU/mL).  This is a clearly reported study overall with defined study population, 
adequate study power, and appropriate statistical methods.   The findings are not surprising and 
largely support  similar studies in other cohorts, but nevertheless provide stronger support to a 
growing body of literature linking quantitative HBsAg levels both at baseline, on -treatment, and 
end-of-treatment to longer term HBsAg loss outcomes.  
 
 
ADDITIONAL REQUESTS/SUGG ESTIONS:  
 
Specific comments:  
 
1) Methods, study population: although clarified in Figure 1, would suggest clarifying that the 
2451 patients were all HBsAg+ patients who received IFN -based therapy  
 
2) Methods, study population: please define "with antiviral t herapy for at least 3 months" -- does 
this refer to IFN -based therapies only? were some of these patients concurrently on oral 
nucleos(t)ide analogues?  
 
3) Methods: did the authors consider analysis of HBeAg negative patients in this study?  
although a dist inct population from HBeAg positive, this would have contributed novel 
information  
 
4) Methods, data collection: the variables collected are remarkably limited for a study of this 
nature - a number of key variables appear to be missing (e.g. cirrhosis/no ci rrhosis)  
 
5) Methods: the authors do not define "IFN -based therapy" - does this include standard IFN -
alpha only, IFN -alpha and peg -IFN alpha?  
 
6) Statistical analysis: please clarify "different EOT HBsAg levels" -- did the authors consider 
other thresholds t han those reported in the results section?  
 
7) Results, patient characteristics (including Table 1): do you have information on HBV 
genotype, HBeAg titer, liver fibrosis stage, treatment experience with prior IFN or nucs)  
 
8) Results: although the focus of t his analysis is on EOT HBsAg levels, and the authors do report 
baseline and wk 24 HBsAg levels, it would also be useful to report on -treatment  wk 12 HBsAg 
levels if available (ok if not available)  
 
9) Results: although implied, the authors should specifica lly include a definition for "HBsAg 
loss" based on specific assay cut -offs - either here in results and/or in the methods section  
 
10) 
 
 
 
Are the methods appropriate and well described?  
If not, please specify what 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 manuscript or would you recommend an  
additional statist ical review?  
If an additional statistical review is recommended, please specify what aspects require further  
assessment in your comments to the editors.  
 
I am able to assess the statistics  
 
Quality of written English  
Please indicate the quality of language i n 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 reimbursement s, fees, funding, or salary from an  
organisation that may in any way gain or lose financially from the publication of this  
manuscript, either now or in the future?  
 
2. Do you hold any stocks or shares in an organisation that 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 currently 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 al l 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. R eviewers 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 completing 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 understand that my nam e will be included  
on my report to the authors and, if the manuscript is accepted for publication, my named report  
including any attachments I upload will be posted on the website along with the authors'  
responses. I agree for my report to be made availabl e 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 included 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: HBsAg quantification predicts off -treatment response to interferon in chronic hepatitis B  
patients：A retrospective study of 250 cases  
 
Version: 0 Date: 29 Jan 2020  
 
Reviewer: Research Square Reviewer 1  
 
Reviewer's report:  
 
PEER REVIEWER ASSESSMENTS:  
 
OBJECTIV E - Full research articles: is there a clear objective that addresses a testable research 
question(s) (brief or other article types: is there a clear objective)?  
Yes - there is a clear objective  
 
DESIGN - Is the current approach (including controls and ana lysis protocols) appropriate for the 
objective?  
Yes - the approach is appropriate  
 
EXECUTION - Are the experiments and analyses performed with technical rigor to allow 
confidence in the results?  
Yes - experiments and analyses were performed appropriately  
 
STATISTICS - Is the use of statistics in the manuscript appropriate?  
Yes - appropriate statistical analyses have been used in the study  
 
INTERPRETATION - Is the current interpretation/discussion of the results reasonable and not 
overstated?  
Yes - the autho r's interpretation is reasonable  
 
OVERALL MANUSCRIPT POTENTIAL - Is the current version of this work technically 
sound? If not, can revisions be made to make the work technically sound?  
Probably - with minor revisions  
 
 
PEER REVIEWER COMMENTS:  
 
GENERAL COM MENTS:  
 
In this manuscript, the authors report the results of a single -center, retrospective cohort study 
which aimed to examine the predictive value of end -of-treatment quantitative HBsAg titer in 
identifying patients who are likely to achieve off -treatme nt HBsAg loss in response to interferon 
therapy.  Using a cohort of 250 patients who underwent IFN therapy for chronic HBV infection 
between 2010 -2017, EOT HBsAg level &lt;10 IU/mL was associated with 30.4% cumulative 
HBsAg loss rate 5 years post -treatment , compared with 9.8% (HBsAg 10 -200 IU/mL) or 0% 
(HBsAg &gt;200 IU/mL).  This is a clearly reported study overall with defined study population, 
adequate study power, and appropriate statistical methods.   The findings are not surprising and 
largely support  similar studies in other cohorts, but nevertheless provide stronger support to a 
growing body of literature linking quantitative HBsAg levels both at baseline, on -treatment, and 
end-of-treatment to longer term HBsAg loss outcomes.  
 
 
ADDITIONAL REQUESTS/SUGG ESTIONS:  
 
Specific comments:  
 
1) Methods, study population: although clarified in Figure 1, would suggest clarifying that the 
2451 patients were all HBsAg+ patients who received IFN -based therapy  
 
2) Methods, study population: please define "with antiviral t herapy for at least 3 months" -- does 
this refer to IFN -based therapies only? were some of these patients concurrently on oral 
nucleos(t)ide analogues?  
 
3) Methods: did the authors consider analysis of HBeAg negative patients in this study?  
although a dist inct population from HBeAg positive, this would have contributed novel 
information  
 
4) Methods, data collection: the variables collected are remarkably limited for a study of this 
nature - a number of key variables appear to be missing (e.g. cirrhosis/no ci rrhosis)  
 
5) Methods: the authors do not define "IFN -based therapy" - does this include standard IFN -
alpha only, IFN -alpha and peg -IFN alpha?  
 
6) Statistical analysis: please clarify "different EOT HBsAg levels" -- did the authors consider 
other thresholds t han those reported in the results section?  
 
7) Results, patient characteristics (including Table 1): do you have information on HBV 
genotype, HBeAg titer, liver fibrosis stage, treatment experience with prior IFN or nucs)  
 
8) Results: although the focus of t his analysis is on EOT HBsAg levels, and the authors do report 
baseline and wk 24 HBsAg levels, it would also be useful to report on -treatment  wk 12 HBsAg 
levels if available (ok if not available)  
 
9) Results: although implied, the authors should specifica lly include a definition for "HBsAg 
loss" based on specific assay cut -offs - either here in results and/or in the methods section  
 
10) 
 
 
 
Are the methods appropriate and well described?  
If not, please specify what 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 manuscript or would you recommend an  
additional statist ical review?  
If an additional statistical review is recommended, please specify what aspects require further  
assessment in your comments to the editors.  
 
I am able to assess the statistics  
 
Quality of written English  
Please indicate the quality of language i n 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 reimbursement s, fees, funding, or salary from an  
organisation that may in any way gain or lose financially from the publication of this  
manuscript, either now or in the future?  
 
2. Do you hold any stocks or shares in an organisation that 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 currently 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 al l 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. R eviewers 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 completing 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 understand that my nam e will be included  
on my report to the authors and, if the manuscript is accepted for publication, my named report  
including any attachments I upload will be posted on the website along with the authors'  
responses. I agree for my report to be made availabl e 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 included 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: HBsAg quantification predicts off -treatment response to interferon in chronic hepatitis B  
patients：A retrospective study of 250 cases  
 
Version: 0 Date: 29 Jan 2020  
 
Reviewer: Research Square Reviewer 1  
 
Reviewer's report:  
 
PEER REVIEWER ASSESSMENTS:  
 
OBJECTIV E - Full research articles: is there a clear objective that addresses a testable research 
question(s) (brief or other article types: is there a clear objective)?  
Yes - there is a clear objective  
 
DESIGN - Is the current approach (including controls and ana lysis protocols) appropriate for the 
objective?  
Yes - the approach is appropriate  
 
EXECUTION - Are the experiments and analyses performed with technical rigor to allow 
confidence in the results?  
Yes - experiments and analyses were performed appropriately  
 
STATISTICS - Is the use of statistics in the manuscript appropriate?  
Yes - appropriate statistical analyses have been used in the study  
 
INTERPRETATION - Is the current interpretation/discussion of the results reasonable and not 
overstated?  
Yes - the autho r's interpretation is reasonable  
 
OVERALL MANUSCRIPT POTENTIAL - Is the current version of this work technically 
sound? If not, can revisions be made to make the work technically sound?  
Probably - with minor revisions  
 
 
PEER REVIEWER COMMENTS:  
 
GENERAL COM MENTS:  
 
In this manuscript, the authors report the results of a single -center, retrospective cohort study 
which aimed to examine the predictive value of end -of-treatment quantitative HBsAg titer in 
identifying patients who are likely to achieve off -treatme nt HBsAg loss in response to interferon 
therapy.  Using a cohort of 250 patients who underwent IFN therapy for chronic HBV infection 
between 2010 -2017, EOT HBsAg level &lt;10 IU/mL was associated with 30.4% cumulative 
HBsAg loss rate 5 years post -treatment , compared with 9.8% (HBsAg 10 -200 IU/mL) or 0% 
(HBsAg &gt;200 IU/mL).  This is a clearly reported study overall with defined study population, 
adequate study power, and appropriate statistical methods.   The findings are not surprising and 
largely support  similar studies in other cohorts, but nevertheless provide stronger support to a 
growing body of literature linking quantitative HBsAg levels both at baseline, on -treatment, and 
end-of-treatment to longer term HBsAg loss outcomes.  
 
 
ADDITIONAL REQUESTS/SUGG ESTIONS:  
 
Specific comments:  
 
1) Methods, study population: although clarified in Figure 1, would suggest clarifying that the 
2451 patients were all HBsAg+ patients who received IFN -based therapy  
 
2) Methods, study population: please define "with antiviral t herapy for at least 3 months" -- does 
this refer to IFN -based therapies only? were some of these patients concurrently on oral 
nucleos(t)ide analogues?  
 
3) Methods: did the authors consider analysis of HBeAg negative patients in this study?  
although a dist inct population from HBeAg positive, this would have contributed novel 
information  
 
4) Methods, data collection: the variables collected are remarkably limited for a study of this 
nature - a number of key variables appear to be missing (e.g. cirrhosis/no ci rrhosis)  
 
5) Methods: the authors do not define "IFN -based therapy" - does this include standard IFN -
alpha only, IFN -alpha and peg -IFN alpha?  
 
6) Statistical analysis: please clarify "different EOT HBsAg levels" -- did the authors consider 
other thresholds t han those reported in the results section?  
 
7) Results, patient characteristics (including Table 1): do you have information on HBV 
genotype, HBeAg titer, liver fibrosis stage, treatment experience with prior IFN or nucs)  
 
8) Results: although the focus of t his analysis is on EOT HBsAg levels, and the authors do report 
baseline and wk 24 HBsAg levels, it would also be useful to report on -treatment  wk 12 HBsAg 
levels if available (ok if not available)  
 
9) Results: although implied, the authors should specifica lly include a definition for "HBsAg 
loss" based on specific assay cut -offs - either here in results and/or in the methods section  
 
10) 
 
 
 
Are the methods appropriate and well described?  
If not, please specify what 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 manuscript or would you recommend an  
additional statist ical review?  
If an additional statistical review is recommended, please specify what aspects require further  
assessment in your comments to the editors.  
 
I am able to assess the statistics  
 
Quality of written English  
Please indicate the quality of language i n 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 reimbursement s, fees, funding, or salary from an  
organisation that may in any way gain or lose financially from the publication of this  
manuscript, either now or in the future?  
 
2. Do you hold any stocks or shares in an organisation that 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 currently 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 al l 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. R eviewers 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 completing 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 understand that my nam e will be included  
on my report to the authors and, if the manuscript is accepted for publication, my named report  
including any attachments I upload will be posted on the website along with the authors'  
responses. I agree for my report to be made availabl e 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 included as confidential comments  
to the editors, which will not be published.  
 
I agree to the open peer review policy of the journal  
assistant