Reviewer ’s report  
Title:  Peripheral blood toll -like receptor 4 correlates with rapid virological response to 
Pegylatedinterferon and ribavirin therapy in hepatitis C genotype 1 patients  
 
Version:  0 Date: 17 Jan 2016  
 
Reviewer:  Vincenzo  Palmieri  
 
Reviewer's report:  
Main finding of this work is that mRNA expression of TLR 2, 3, 4, 7, and 9 was lower in the 
RVR group, compared with the non -RVR group. This indicates that TLR (TLR4 in particolar) 
may play  a crucial role in HCV recognition and activation.  
Bu the authors do not explain why the relation between mRNA expression of these receptors and 
the SVR (that represents the real therapeuic aim of the PG -IFN/RBV therapy) is absent.  
They do not spend any wo rd in the discussion in order to explain this great limit of the study.  
Moreover, they correctly mention the work of He et al (Liver International 2006), that has some 
methodological overlapping with this work, but in which the levels of expression of TLRs  
receptors are evacuate in relation to the SVR and not to the RVR.  He et al. that baseline PBMC 
TLR 2 and 3 mRNA levels were significantly higher in patients with SVR than in non -
responders, a result that is opposite to that reported by the authors of the  proposed paper in this 
review. Even if in the paper of Liver International the number of patients is low, the authors need 
to analyze with more details the reasons of this so clear discrepant conclusion. This topic is 
absolutely relevant and I wish to men tion a "small" but more recent paper published in chinese 
language (so I could have access only to the abstract) (see complete enclosed reference): the 
authors show that surface expression of PD -1, TLR4, and PD -1:TLR4 is up -regulated in the total 
PBMCs of CHC patients and that Peg -IFNalpha -2a plus RBV treatment -induced suppression of 
HCV replication results in a significant reduction in PD -1 and TLR4 expression on the surface of 
PBMCs. This data seems to confirm the results of the work of He et al, but not with that of the 
paper we are reviewing.  
The auhors, therefore, need to explain:  
- the reasons why the relation of mRNA expressione of TLRs with the RVR is lost when the 
SVR is taken into account  
- the reasons why their resuls is in contrast with the twuo previous work that I have mentioned  
 
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.  
No 
Are you able to assess any statist ics in the manuscript or would you recommend an 
additional statistical 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 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 
organi sation that may in any way gain or lose financial ly from the publication 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 manus cript, 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 pat ents 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.  
I declare that I ha ve no competing interests  
I agree to the open peer review policy of the journal. I understand that my name 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 wi ll 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 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:  Peripheral blood toll -like receptor 4 correlates with rapid virological response to 
Pegylatedinterferon and ribavirin therapy in hepatitis C genotype 1 patients  
 
Version:  0 Date: 17 Jan 2016  
 
Reviewer:  Vincenzo  Palmieri  
 
Reviewer's report:  
Main finding of this work is that mRNA expression of TLR 2, 3, 4, 7, and 9 was lower in the 
RVR group, compared with the non -RVR group. This indicates that TLR (TLR4 in particolar) 
may play  a crucial role in HCV recognition and activation.  
Bu the authors do not explain why the relation between mRNA expression of these receptors and 
the SVR (that represents the real therapeuic aim of the PG -IFN/RBV therapy) is absent.  
They do not spend any wo rd in the discussion in order to explain this great limit of the study.  
Moreover, they correctly mention the work of He et al (Liver International 2006), that has some 
methodological overlapping with this work, but in which the levels of expression of TLRs  
receptors are evacuate in relation to the SVR and not to the RVR.  He et al. that baseline PBMC 
TLR 2 and 3 mRNA levels were significantly higher in patients with SVR than in non -
responders, a result that is opposite to that reported by the authors of the  proposed paper in this 
review. Even if in the paper of Liver International the number of patients is low, the authors need 
to analyze with more details the reasons of this so clear discrepant conclusion. This topic is 
absolutely relevant and I wish to men tion a "small" but more recent paper published in chinese 
language (so I could have access only to the abstract) (see complete enclosed reference): the 
authors show that surface expression of PD -1, TLR4, and PD -1:TLR4 is up -regulated in the total 
PBMCs of CHC patients and that Peg -IFNalpha -2a plus RBV treatment -induced suppression of 
HCV replication results in a significant reduction in PD -1 and TLR4 expression on the surface of 
PBMCs. This data seems to confirm the results of the work of He et al, but not with that of the 
paper we are reviewing.  
The auhors, therefore, need to explain:  
- the reasons why the relation of mRNA expressione of TLRs with the RVR is lost when the 
SVR is taken into account  
- the reasons why their resuls is in contrast with the twuo previous work that I have mentioned  
 
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.  
No 
Are you able to assess any statist ics in the manuscript or would you recommend an 
additional statistical 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 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 
organi sation that may in any way gain or lose financial ly from the publication 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 manus cript, 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 pat ents 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.  
I declare that I ha ve no competing interests  
I agree to the open peer review policy of the journal. I understand that my name 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 wi ll 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 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:  Peripheral blood toll -like receptor 4 correlates with rapid virological response to 
Pegylatedinterferon and ribavirin therapy in hepatitis C genotype 1 patients  
 
Version:  0 Date: 17 Jan 2016  
 
Reviewer:  Vincenzo  Palmieri  
 
Reviewer's report:  
Main finding of this work is that mRNA expression of TLR 2, 3, 4, 7, and 9 was lower in the 
RVR group, compared with the non -RVR group. This indicates that TLR (TLR4 in particolar) 
may play  a crucial role in HCV recognition and activation.  
Bu the authors do not explain why the relation between mRNA expression of these receptors and 
the SVR (that represents the real therapeuic aim of the PG -IFN/RBV therapy) is absent.  
They do not spend any wo rd in the discussion in order to explain this great limit of the study.  
Moreover, they correctly mention the work of He et al (Liver International 2006), that has some 
methodological overlapping with this work, but in which the levels of expression of TLRs  
receptors are evacuate in relation to the SVR and not to the RVR.  He et al. that baseline PBMC 
TLR 2 and 3 mRNA levels were significantly higher in patients with SVR than in non -
responders, a result that is opposite to that reported by the authors of the  proposed paper in this 
review. Even if in the paper of Liver International the number of patients is low, the authors need 
to analyze with more details the reasons of this so clear discrepant conclusion. This topic is 
absolutely relevant and I wish to men tion a "small" but more recent paper published in chinese 
language (so I could have access only to the abstract) (see complete enclosed reference): the 
authors show that surface expression of PD -1, TLR4, and PD -1:TLR4 is up -regulated in the total 
PBMCs of CHC patients and that Peg -IFNalpha -2a plus RBV treatment -induced suppression of 
HCV replication results in a significant reduction in PD -1 and TLR4 expression on the surface of 
PBMCs. This data seems to confirm the results of the work of He et al, but not with that of the 
paper we are reviewing.  
The auhors, therefore, need to explain:  
- the reasons why the relation of mRNA expressione of TLRs with the RVR is lost when the 
SVR is taken into account  
- the reasons why their resuls is in contrast with the twuo previous work that I have mentioned  
 
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.  
No 
Are you able to assess any statist ics in the manuscript or would you recommend an 
additional statistical 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 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 
organi sation that may in any way gain or lose financial ly from the publication 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 manus cript, 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 pat ents 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.  
I declare that I ha ve no competing interests  
I agree to the open peer review policy of the journal. I understand that my name 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 wi ll 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 included as confidential comments 
to the editors, which will not be published.  
I agree to the open peer review policy of the journal  
assistant