CX3CL1/Fractalkine: A Potential Biomarker for Liver Fibrosis in Chronic HBV Infection

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

Arsentieva et al. suggested that CX3CL1/Fractalkine may be a prognostic biomarker for the development of liver fibrosis.

The manuscript had interesting findings and is well-written as well, however, there are some comcerns.

1. Schematic diagram of the experimental design is necessary for easier understanding in the current study.

2. L92-102 : The authors should describe in detail with profer references how the patient or normal person was recruited, how/who it was identified as a patient, what category and method the degree of fibrosis was classified, and how it was excluded from the recruitment group.

3. How to did the authors define and categorize HBV and HCB patients? Which categories? In case of patient with chronic hepatitis B, it is defined HBsAg+ > 6 months, serum HBV DNA > 2000IU/ml, liver biopsy showing chronic heatitis, and so on.  Therefore, authors should be presented the exact methods and its results both.

4. In the Table 1, this should include more patient information in detail, as shown in clinical studies involving patients with hepatic fibrosis and viral infection. In addition, statistical analysis should be addressed in all group.

5. In the Table 1, ALT/AST ratio should be addressed.

6. In the Figure 1, analysis using anova statistics should be done to compare group.

7. Multivariate regression analysis should be performed to identify any possible association among clinical feature.

8. In IRB information, current study was approved at 1 Dec 2016, correct? In my understanding, it seems to be the date for the first study for data collection in a study other than the current study. If so, is it possible to apply the first approved IRB to other studies? Research ethics is very strict in clinical research, so please check again.

9. Where is the part of discussion? Please check again.

Author Response

Dear Reviewer,

Thank you for your interest in our manuscript. We have made changes according to your commentary:

1. Schematic diagram of the experimental design is necessary for easier understanding in the current study.

Thank you for the advice. We added Figure 1 representing experimental design.

2. The authors should describe in detail with profer references how the patient or normal person was recruited, how/who it was identified as a patient, what category and method the degree of fibrosis was classified, and how it was excluded from the recruitment group.
3. How to did the authors define and categorize HBV and HCB patients? Which categories? In case of patient with chronic hepatitis B, it is defined HBsAg+ > 6 months, serum HBV DNA > 2000IU/ml, liver biopsy showing chronic hepatitis, and so on.  Therefore, authors should be presented the exact methods and its results both.

Thank you. We added all available information on the patients included within the study, including criteria for diagnosis.

4. In the Table 1, this should include more patient information in detail, as shown in clinical studies involving patients with hepatic fibrosis and viral infection. In addition, statistical analysis should be addressed in all group.
5. In the Table 1, ALT/AST ratio should be addressed.

Thank you for your feedback. We have included more details about the patients in the text and added the AST/ALT ratio. We would appreciate it if the Reviewer could clarify what type of statistical analysis they are referring to in Table 1. We would also like to remind the Reviewer that this table presents the demographics of the cohort and is not part of the Results section.

6. In the Figure 1, analysis using anova statistics should be done to compare group.

Thank you for your commentary. We changed Figure 1 (now Figure 2), and modified the analysis to improve the clarity of comparisons. We used the Kruskal-Wallis test for multivariable analysis at the same fibrosis stage and used the Mann-Whitney test specifically for HBV.

7. Multivariate regression analysis should be performed to identify any possible association among clinical feature.

Thank you for your commentary. We performed multivariate regression analysis as you suggested. Unfortunately, we did not receive any statistically significant results for the groups with different fibrosis. We believe this may be due to the little impact that ALT and AST have on the fibrosis stage, as their increase is a consequence of the liver damage, not vice versa. Other factors, such as age and gender, also do not seem to contribute to the severity of fibrosis. Therefore, the results of the regression analysis were not satisfactory for publication.

8. In IRB information, current study was approved at 1 Dec 2016, correct? In my understanding, it seems to be the date for the first study for data collection in a study other than the current study. If so, is it possible to apply the first approved IRB to other studies? Research ethics is very strict in clinical research, so please check again.

Thank you for your observation. This research is part of a broader study that was covered by various ethics committee approvals. However, we have included extra information regarding the more relevant ethics committee declaration.

9. Where is the part of discussion? Please check again.

Thank you for your observation. The CIMB editors recommended that we move this article from another journal that doesn’t require a Discussion section in its Communication format. We revised the Results section and included additional sources to create a comprehensive Discussion.

Thank you again for the time and effort put into reviewing our manuscript.

Reviewer 2 Report

Comments and Suggestions for Authors

Dear authors

 

The paper is interesting but there are some limitations that must be highlighted: 

The paper includes only 47 patients which limits a lot of conclusions - this fact might be in study limitations

In patients with HBV and low grade of fibrosis, how were the levels of CXCL1? Can you conclude that CXCL1 would be an important maker for fibrosis development? Or that only appears in advanced rates of fibrosis? In this last case it would be not so important.

 

Do you have any information about HCC? Liver decompensation in patients according to CXCL1 levels?

 

There should be a specific place for discussion and not only results and conclusion.

 

Author Response

Dear Reviewer, thank you kindly for the time and effort put into reading our manuscript.

The paper includes only 47 patients which limits a lot of conclusions - this fact might be in study limitations

The small sample size is a result of the challenges in obtaining blood from patients with acute-stage chronic hepatitis, and we have noted this as a limitation of the study.

In patients with HBV and low grade of fibrosis, how were the levels of CXCL1? Can you conclude that CXCL1 would be an important maker for fibrosis development? Or that only appears in advanced rates of fibrosis? In this last case it would be not so important.

As illustrated in Figure 2 (previously Figure 1), CX3CL1 levels decrease even in the early stages of fibrosis. We have emphasized this further in the Discussion section.

Do you have any information about HCC? Liver decompensation in patients according to CXCL1 levels?

Thank you for your inquiry. To illustrate liver dysfunction, we included data on the blood enzyme ratio (AST/ALT) in the patient cohort description. This ratio is commonly used in clinical practice to indicate liver function decompensation and cell damage during fibrosis or cirrhosis. Additionally, fibrosis development observed through elastography serves as another, albeit secondary, marker of liver dysfunction.

There should be a specific place for discussion and not only results and conclusion.

 Thank you for your observation. The CIMB editors recommended that we move this article from another journal that doesn’t require a Discussion section in its Communication format. We revised the Results section and included additional sources to create a comprehensive Discussion.

Thank you again for your review. We hope you find changes satisfactory.

Round 2

Reviewer 1 Report

Comments and Suggestions for Authors

Authors addressed all my concerns.

Therefore, it seems that the current revised manuscript could be sufficiently published in the CIME.

Reviewer 2 Report

Comments and Suggestions for Authors

Dear authors

 

I suggest this paper to be transferred to another journal with lower impact factor as this manuscript has some methodological errors that cannot be surpassed.