Impact of IL-12B Genetic Variants on Antiviral Treatment Response among Hepatitis B Patients in Pakistan

Round 1

Reviewer 1 Report

This article described the role of IL-12B Genetic Variants in the immune response in viral therapy for Pakistan hepatitis B patients. The authors divided and compared three groups ( SVR, NSV, fresh) and found that the variant genotype CC was significantly associated with the NSV to the antiviral therapy. It is a well-written and exciting article. However, there are some concerns about this article. 1. The title should include the population name (e.g., Pakistan HBV patients) since several similar studies exist in other countries. 2. The authors should explain to determine the normal value of ALT as below 40 IU/L, not 30 IU/L. 3. In the article, please delete the reference name(e.g., Masood et al., 2019) in the text and add these in the reference section. The number of references is lacking(e.g., 1. 2. ) 3. The authors could present these HBV-infected patients' demographic data tables.

 

 

 

 

Moderate English editing is required. 

Author Response

Comments and Suggestions for Authors

This article described the role of IL-12B Genetic Variants in the immune response in viral therapy for Pakistan hepatitis B patients. The authors divided and compared three groups ( SVR, NSV, fresh) and found that the variant genotype CC was significantly associated with the NSV to the antiviral therapy. It is a well-written and exciting article. However, there are some concerns about this article.

We are thankful to the reviewer for their positive feedback and valuable suggestions on our manuscript. These suggestions have made authors feel encouraged and motivated to improve the manuscript. We have discussed all the comments point by point as follows:

 

1). The title should include the population name (e.g., Pakistan HBV patients) since several similar studies exist in other countries.

We are thankful for this valuable and thoughtful suggestion. We have mentioned in our manuscript that this study is focused on the Pakistani population as it exclusively involved HBV patients and control participants from Pakistan. However, we have modified the title of this manuscript by the addition of a phrase “in Pakistan” to “Impact of IL-12B Genetic Variants on Antiviral Treatment Response among Hepatitis B Patients in Pakistan”.

 

2). The authors should explain to determine the normal value of ALT as below 40 IU/L, not 30 IU/L.

We appreciate this insightful comment from the reviewer regarding the normal levels of ALT.

In response to your valuable comment, there are several studies that have reported that the normal levels of ALT can range from 7-44 IU/ml. The levels of ALT can be affected by various factors such as diet,  lifestyle, alcohol consumption etc. and all these factors tend to impact an individual's ALT. Moreover, studies have reported that in Pakistan the upper lower normal (ULN) for ALT levels is up to 40 IU/ml in the healthy population. therefore, in this study we selected normal levels of ALT below 40 IU/ml instead of 30 IU/ml.

3). In the article, please delete the reference name(e.g., Masood et al., 2019) in the text and add these in the reference section. The number of references is lacking (e.g., 1. 2. )

We are thankful for this suggestion from the reviewer. We have modified the style of in-text citations in this manuscript and the revised version of our manuscript has references in numbered style.

4). The authors could present these HBV-infected patients' demographic data tables.

We are thankful for this suggestion. We have mentioned the participant's demographic data in the tabular form in the results section (Table 1).

Reviewer 2 Report

The research article by Badshah et al., entitled “Impact of IL-12B Genetic Variants on Antiviral Treatment Response among Hepatitis B Patients” examined impact of interleukins 12 polymorphism on response to antiviral treatment among hepatitis B virus infected patients.

I have following major concerns about the study.

1) Authors need to mention the definition of sustained virological response, NSR and fresh cases in details, and all the parameters taken for consideration for exclusion and inclusion criteria also need to be mentioned in detail in the materials and method section. For example, like what was the cutoff for HBV DNA, liver function test, serological markers like HBsAg, HBeAg detail. did the authors followed any guidelines like American association for study of liver disease or European Association for study of liver disease or Asian pacific association for classifying the patients?

2) Did the control cases were age sex matched? Need to mention in detail.

3) What kind of antiviral therapy was given to patients? What was the dose and duration of therapies? All these details need to include in materials and methods section.

4) Page 2, line 86, please write the full form of ASAB and NUST.

5) Page 2, line 91, The HBV-positive samples were categorized into three groups sustained virological responders (NSR) which were positive for HBV DNA after. The underlined phrase is not correct, it says sustained virological responder but in the bracket is written NSR.

6) In the statistical analysis section, please mention the methods which was used to calculate p value.

7) Figure 1, please plot the bar graph with SE/SD and p value significance between the groups.

8) What could be the reason for low level of ALT in the SVR group as compared to healthy controls (Figure-1). This needs to explain.

9) Page 5, line 184, 440+8015873 184 IU/ml.  The DNA value need to recheck because SE value is more.

10) Figure 2A, please plot the bar graph with SE/SD and p value significance between the groups.

11) Figure 2B, please plot the bar graph with p value significance between the groups.

12) For the detection of HBV DNA, authors have used nested PCR approach, authors should include gel photograph in the result section.

13) Conclusions need to be more concise and focused.

14) Some of the sentences need proper English grammar correction and rephrasing.

 

 

 

 

 

Some of the sentences need proper English grammar correction and rephrasing.

Author Response

The research article by Badshah et al., entitled “Impact of IL-12B Genetic Variants on Antiviral Treatment Response among Hepatitis B Patients” examined impact of interleukins 12 polymorphism on response to antiviral treatment among hepatitis B virus infected patients.

I have following major concerns about the study.

We appreciate the insightful feedback from the reviewer on our manuscript. We have revised and enhanced the manuscript as per the suggestions made by the reviewer by explaining the rationale behind sample selection, elaborating on the genotyping results, and expanding the discussion on the significance of the findings from our research. We have strongly felt that your input is invaluable in refining the quality of our research.

We have explained all the suggestions and comments from the reviewer in our revised manuscript and discussed them point-by-point as follows:

1)  Authors need to mention the definition of sustained virological response, NSR and fresh cases in details, and all the parameters taken for consideration for exclusion and inclusion criteria also need to be mentioned in detail in the materials and method section. For example, like what was the cutoff for HBV DNA, liver function test, serological markers like HBsAg, HBeAg detail.

Thank you for your meticulous review of our manuscript. Your suggestions for improving the clarity and comprehensiveness of our Materials and Methods section are greatly appreciated. We have received the manuscript and have made sure to incorporate a comprehensive description of the suggested elements including the definition of study groups such as NSR, SVR, Fresh cases, and control in our methodology section. Furthermore, have also included the exclusion and inclusion criteria that were applied in this study for sample selection. Lastly, we have also included the cut-off values for ALT, HBV DNA, and serological markers in detail, further explaining the sample selection criteria under the methodology section. We have felt that this suggestion has made our manuscript more coherent and clear. we sincerely appreciate your insightful suggestions to improve the overall quality of our manuscript.

 

2) Did the authors followed any guidelines like American association for study of liver disease or European Association for study of liver disease or Asian pacific association for classifying the patients? Did the control cases were age sex matched? Need to mention in detail.

We appreciate the insightful comment from the reviewer. The patient samples in this study were processed and categorized as per the guidelines of the American Association for the Study of Liver Diseases (AASLD). We have also mentioned this critical information in the methodology section of our revised manuscript. We also performed the age and sex matching in order to avoid the confounding effect in this study to maintain the authenticity of the generated data. We are grateful for the thoughtful review of our manuscript.

 

3) What kind of antiviral therapy was given to patients? What was the dose and duration of therapies? All these details need to include in materials and methods section.

We are immensely grateful to the reviewer for bringing this crucial point to our notice. We have revised the manuscript and in the modified version we have mentioned the antiviral therapy, its dosage and duration for the treatment of HBV patients in the methodology section. We have felt that this addition of information has made our manuscript more understandable.

 

4) Page 2, line 86, please write the full form of ASAB and NUST.

Thank you for this comment. We have mentioned the full form of ASAB in the methodology section as per the advise of the reviewer.

5) Page 2, line 91, The HBV-positive samples were categorized into three groups sustained virological responders (NSR) which were positive for HBV DNA after. The underlined phrase is not correct, it says sustained virological responder but in the bracket is written NSR.

We apologize for this mistake and we are also thankful for the interest of and thoughtful comments by the reviewer to make our manuscript error-proof and understandable. We have rectified this error in the methodology section.

6) In the statistical analysis section, please mention the methods which was used to calculate p value.

We are thankful for this suggestion by the reviewer. We already had mentioned the statistical analysis that was used i.e. fisher exact test, to determine the significant p-value for this study in the last section of methodology. However, as per the advice, we have modified this section and have further elaborated on the methodology that was used to determine the P-value, Odds ratio, relative risk and 95%CI.

7) Figure 1, please plot the bar graph with SE/SD and p value significance between the groups.

We are grateful to the reviewer for their advice. We have modified the figure 1 and have included the error lines and significant p-value among the study groups in the revised version of our manuscript.

8) What could be the reason for low level of ALT in the SVR group as compared to healthy controls (Figure-1). This needs to explain.

We appreciate your insightful feedback and for raising a valid point regarding the lower levels of ALT observed in the SVR group as compared to the healthy controls, as depicted in Figure-1. There are several reasons that are responsible for this difference in the ALT values which include the lifestyle such as a high-fat and high-sugar diet, and mild to high alcohol consumption that can affect the levels of ALT levels in individuals with no reported health conditions. While in the HBV-positive patients with SVR, the therapy they received normalized their ALT levels which were even lower compared to those of controls.  We have elaborated on the possible reasons that explain this discrepancy of the ALT levels in the discussion section as well.  

9) Page 5, line 184, 440+8015873 184 IU/ml.  The DNA value need to recheck because SE value is more.

We are thankful to the reviewer for bringing this to our attention. We have rectified this mistake and in the revised version of our manuscript, we have mentioned the correct value of viral load in the SVR group.

10) Figure 2A, please plot the bar graph with SE/SD and p value significance between the groups.  

11) Figure 2B, please plot the bar graph with p value significance between the groups.

We are thankful for this valuable suggestion. We have modified the figure 2a by plotting the error lines for each group. Moreover, we have also represented the p-value significance for viral load among the study groups. We have also modified Figure 2b and have presented the p-value difference. It is important to mention here that the p-value difference among the three genotype groups was not significant. However, we have still presented the p-value line on the bar graph.

 

12) For the detection of HBV DNA, authors have used a nested PCR approach, authors should include a gel photograph in the result section.

We are thankful to the reviewer for their valuable feedback. We apologize for this mistake. HBV nested PCR was performed in this study.  We have revised the manuscript and included the picture of the HBV nested PCR gel in the results section as per the advise of the reviewer.

 

13) Conclusions need to be more concise and focused.

As per the advice of the reviewer, we have modified the conclusion section of our manuscript to make it more concise and focused on the main finding of this study. we appreciate the reviewer for their insightful comment.

 

14) Some of the sentences need proper English grammar correction and rephrasing.

We appreciate the sincere feedback from the reviewer. We understand the significance of maintaining clear and effective communication therefore We have revised the manuscript to address these errors related to grammar and sentence structure and ensured that the language meets the standards expected for publication.

Round 2

Reviewer 2 Report

I have no further questions.