Anti-HCV antibody titer highly predicts HCV viremia in patients with hepatitis B virus dual-infection

Hung-Yin Liu; Yi-Hung Lin; Pei-Ju Lin; Pei-Chien Tsai; Shu-Fen Liu; Ying-Chou Huang; Jia-Jiun Tsai; Ching-I Huang; Ming-Lun Yeh; Po-Cheng Liang; Zu-Yau Lin; Chia-Yen Dai; Jee-Fu Huang; Wan-Long Chuang; Chung-Feng Huang; Ming-Lung Yu

doi:10.1371/journal.pone.0254028

Peer Review History

Original SubmissionApril 29, 2021
17 May 2021 Decision Letter - Tatsuo Kanda, Editor PONE-D-21-14163 Anti-HCV antibody titer highly predicts HCV viremia in patients with hepatitis B virus dual-infection PLOS ONE Dear Dr. Hung Yin Liu, Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process. Please submit your revised manuscript by Jun 27 2021 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org. When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file. 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If the funders had no role in your study, please state: “The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.” If any authors received a salary from any of your funders, please state which authors and which funders. If you did not receive any funding for this study, please state: “The authors received no specific funding for this work.” Please include your amended statements within your cover letter; we will change the online submission form on your behalf. [Note: HTML markup is below. Please do not edit.] Reviewers' comments: Reviewer's Responses to Questions Comments to the Author 1. Is the manuscript technically sound, and do the data support the conclusions? The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented. Reviewer #1: Partly Reviewer #2: Partly ******** 2. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: Yes Reviewer #2: I Don't Know ****** 3. Have the authors made all data underlying the findings in their manuscript fully available? The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified. Reviewer #1: Yes Reviewer #2: Yes ****** 4. Is the manuscript presented in an intelligible fashion and written in standard English? PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here. Reviewer #1: Yes Reviewer #2: Yes ****** 5. Review Comments to the Author Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters) Reviewer #1: 1. Please mention if patients had history of anti-HBV therapy including interferon therapy. 2. As for patients with dual-infection, did all have HBV DNA positive? Please mention if there were any patient who had HBV DNA negative. Is there difference in HCV viral load between mono-infected and dual-infected patients? 3. Please mention whether all patients with HCV viremia were confirmed to have persistent infection. 4. Please provide HCV RNA viral load according to the anti-HCV antibody titer. 5. Please mention to what extent the different method of anti-HCV measurement affects the anti-HCV titer and cut-off value according to the previous reports. 6. In the discussion, please elaborate factors affecting low anti-HCV titer with HCV viremia or high anti-HCV titer with HCV negativity. 7. The method of HCV genotyping is not described. Reviewer #2: In the manuscript titled “ Anti-HCV antibody titer highly predicts HCV viremia in patients with hepatitis B virus dual-infection”, the authors showed the valuable titer to predict Hepatitis C viremia in patiets with HCV mono-infection. The authors defined HCV infection as HCV-Ab positive. I think that the definition of patients with HBV infection should include not only HBsAg positive but also HBsAb or HBcAb positive. There are patients whose HBsAg are negative but HBV-DNA are positive. HBsAg negative cannot exclude HBV infection. ****** 6. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files. If you choose “no”, your identity will remain anonymous but your review may still be made public. Do you want your identity to be public for this peer review?** For information about this choice, including consent withdrawal, please see our Privacy Policy. Reviewer #1: No Reviewer #2: No [NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.] While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/. PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email PLOS at figures@plos.org. Please note that Supporting Information files do not need this step. https://doi.org/10.1371/journal.pone.0254028.r001
Revision 1
24 May 2021 Author Response Reviewer #1: 1. Please mention if patients had history of anti-HBV therapy including interferon therapy. Reply: Of the 125 HBV dually infected patients, only 3 (2.4 %) underwent nucleoside/nucleotide analogues. Since the case number was limited, it shall not have impact on the result regarding the issue. We have added it in the result of the revised manuscript. Thank you. 2. As for patients with dual-infection, did all have HBV DNA positive? Please mention if there were any patient who had HBV DNA negative. Is there difference in HCV viral load between mono-infected and dual-infected patients? Reply: Thank you for the comment. Of the 125 HBV dually infected patients, forty-nine (39.2 %) had detectable HBV DNA (lower detection limit: 20 IU/mL). The HCV RNA level did not differ between patients with HCV monoinfection and HBV dual infection (5.6 log IU/mL vs. 5.5 log IU/mL, P=0.42). We have denoted it in the revised table 1 accordingly. 3. Please mention whether all patients with HCV viremia were confirmed to have persistent infection. Reply: Thank you for the great comment. The study was executed in the outpatient departments of the medical center. We only tested HCV RNA once for patients who were seropositive for anti-HCV. Most of the patients who were confirmed HCV viremia would be referred and allocated to antiviral therapy. We did not leave the viremic patients being observed and recheck HCV RNA at least 6 months apart thereafter. As mentioned in the text, we may fail to identify and exclude patients with acute hepatitis C infection. However, all the blood samples were retrieved from outpatient clinics during daily practice where acute hepatitis was less likely to be encountered. We have denoted it as the potential limitation in the current study. Thank you again for the comment. 4. Please provide HCV RNA viral load according to the anti-HCV antibody titer. Reply: Among the 1196 HCV viremic patients, the HCV RNA levels were 3.0 log IU/mL, 5.4 log IU/mL, 5.6 log IU/mL and 5.6 log IU/mL in patients whose anti-HCV antibody titer were < 5 S/CO, 5-10 S/CO, 10-15 S/CO and > 15 S/CO, respectively (trend P=0.13). We have denoted it in the revised manuscript. 5. Please mention to what extent the different method of anti-HCV measurement affects the anti-HCV titer and cut-off value according to the previous reports. Reply: Thank you very much for the insightful comment. We fully agree with the review that different anti-HCV testing methods (ex. ELISA, CMIA or MEIA) may result in different cut-off values and accuracy in predicting HCV viremia. We have summarized previous reports and the current study in the supplementary Table 3. Notably, the current study was in line with the report by Seo et al [1]. who used the same chemiluminescent microparticle immunoassay and resulted in a similar cut-off value and accuracy. 6. In the discussion, please elaborate factors affecting low anti-HCV titer with HCV viremia or high anti-HCV titer with HCV negativity. Reply: Thank you for the great comment. As shown in the original Table 3, patients with the extreme mismatch between anti-HCV titer and viremic status were rare. We further analyzed the characteristics of the patients. As shown in Supplementary Table 1, patients (n=3) with anti-HCV S/CO <5 but HCVRNA (+) were older than the counterpart patients. By contrast, patients (n=3) with anti-HCV >10 S/CO but HCVRNA (-) had lower levels of aspartate aminotransferase, alanine aminotransferase and gamma-glutamyl transferase compared to the counterpart patients (supplementary Table 2). We have denoted in the revised manuscript. Thank you again. 7. The method of HCV genotyping is not described. Reply: HCV genotyping was measured using a standardized automated qualitative reverse transcription-polymerase chain reaction (RT-PCR) assay (Abbott Real-Time HCV Assay in m2000 PCR System). We have denoted it accordingly. Attachments Attachment Submitted filename: Response to Reviewers.docx https://doi.org/10.1371/journal.pone.0254028.r002
10 Jun 2021 Decision Letter - Tatsuo Kanda, Editor PONE-D-21-14163R1 Anti-HCV antibody titer highly predicts HCV viremia in patients with hepatitis B virus dual-infection PLOS ONE Dear Dr. Hung Yin Liu, Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process. Please submit your revised manuscript by Jul 25 2021 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org. When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file. Please include the following items when submitting your revised manuscript: A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'. A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'. An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'. If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter. If applicable, we recommend that you deposit your laboratory protocols in protocols.io to enhance the reproducibility of your results. Protocols.io assigns your protocol its own identifier (DOI) so that it can be cited independently in the future. For instructions see: http://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols. Additionally, PLOS ONE offers an option for publishing peer-reviewed Lab Protocol articles, which describe protocols hosted on protocols.io. Read more information on sharing protocols at https://plos.org/protocols?utm_medium=editorial-email&utm_source=authorletters&utm_campaign=protocols. We look forward to receiving your revised manuscript. Kind regards, Tatsuo Kanda, M.D., Ph.D. Academic Editor PLOS ONE Journal Requirements: Please review your reference list to ensure that it is complete and correct. If you have cited papers that have been retracted, please include the rationale for doing so in the manuscript text, or remove these references and replace them with relevant current references. Any changes to the reference list should be mentioned in the rebuttal letter that accompanies your revised manuscript. If you need to cite a retracted article, indicate the article’s retracted status in the References list and also include a citation and full reference for the retraction notice. [Note: HTML markup is below. Please do not edit.] Reviewers' comments: Reviewer's Responses to Questions Comments to the Author 1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation. Reviewer #1: (No Response) Reviewer #2: All comments have been addressed ******** 2. Is the manuscript technically sound, and do the data support the conclusions? The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented. Reviewer #1: (No Response) Reviewer #2: Yes ****** 3. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: (No Response) Reviewer #2: Yes ****** 4. Have the authors made all data underlying the findings in their manuscript fully available? The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified. Reviewer #1: (No Response) Reviewer #2: Yes ****** 5. Is the manuscript presented in an intelligible fashion and written in standard English? PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here. Reviewer #1: (No Response) Reviewer #2: Yes ****** 6. Review Comments to the Author Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters) Reviewer #1: According to table 1, most HBsAg positive patients did not have detectable HBV DNA level. I wonder if such infection stage frequently occurs in the natural course of HBV infection. How about HBsAg level in these patients? Reviewer #2: (No Response) ****** 7. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files. If you choose “no”, your identity will remain anonymous but your review may still be made public. Do you want your identity to be public for this peer review?** For information about this choice, including consent withdrawal, please see our Privacy Policy. Reviewer #1: No Reviewer #2: No [NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.] While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/. PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email PLOS at figures@plos.org. Please note that Supporting Information files do not need this step. https://doi.org/10.1371/journal.pone.0254028.r003
Revision 2
16 Jun 2021 Author Response Reviewer #1: According to table 1, most HBsAg positive patients did not have detectable HBV DNA level. I wonder if such infection stage frequently occurs in the natural course of HBV infection. How about HBsAg level in these patients? Reply: We regret that HBsAg level was not available in the retrospective study. As mentioned in the discussion, there may exist reciprocally suppressive effect of the two viruses, and HCV often overwhelms HBV in the same host during the natural course.1 The mean HBV DNA level was quite low in the present survey, which was similar to our previous report.2 The rate of detectable HBV DNA was around 40 % in the current study, which was also not far from another community-based3 or hospital-based4 studies regarding HBV/HCV dual infection (<50 %). Patients were consecutively enrolled in the current study and we believed that there would be minimal selection bias. We have denoted it in the 2nd revised version. Thank you again for the great comment. References 1. Raimondo G, Brunetto MR, Pontisso P, et al. Longitudinal evaluation reveals a complex spectrum of virological profiles in hepatitis B virus/hepatitis C virus-coinfected patients. Hepatology 2006;43:100-7. 2. Huang CF, Dai CY, Lee JJ, et al. Hepatitis C viremia interferes serum hepatitis B virus surface antigen and DNA levels in hepatitis B uremics. Hepatol Int 2014;8:224-232. 3. Yu ML, Dai CY, Huang CF, et al. High hepatitis B virus surface antigen levels and favorable interleukin 28B genotype predict spontaneous hepatitis C virus clearance in uremic patients. J Hepatol 2014;60:253-9. 4. Yeh ML, Huang CF, Huang CI, et al. Hepatitis B-related outcomes following direct-acting antiviral therapy in Taiwanese patients with chronic HBV/HCV co-infection. J Hepatol 2020;73:62-71. Attachments Attachment Submitted filename: Response to Reviewers.docx https://doi.org/10.1371/journal.pone.0254028.r004
18 Jun 2021 Decision Letter - Tatsuo Kanda, Editor Anti-HCV antibody titer highly predicts HCV viremia in patients with hepatitis B virus dual-infection PONE-D-21-14163R2 Dear Dr. Hung Yin Liu, We’re pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements. Within one week, you’ll receive an e-mail detailing the required amendments. When these have been addressed, you’ll receive a formal acceptance letter and your manuscript will be scheduled for publication. An invoice for payment will follow shortly after the formal acceptance. To ensure an efficient process, please log into Editorial Manager at http://www.editorialmanager.com/pone/, click the 'Update My Information' link at the top of the page, and double check that your user information is up-to-date. If you have any billing related questions, please contact our Author Billing department directly at authorbilling@plos.org. If your institution or institutions have a press office, please notify them about your upcoming paper to help maximize its impact. If they’ll be preparing press materials, please inform our press team as soon as possible -- no later than 48 hours after receiving the formal acceptance. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information, please contact onepress@plos.org. Kind regards, Tatsuo Kanda, M.D., Ph.D. Academic Editor PLOS ONE Additional Editor Comments (optional): Reviewers' comments: Reviewer's Responses to Questions Comments to the Author 1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation. Reviewer #1: (No Response) ******** 2. Is the manuscript technically sound, and do the data support the conclusions? The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented. Reviewer #1: (No Response) ****** 3. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: (No Response) ****** 4. Have the authors made all data underlying the findings in their manuscript fully available? The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified. Reviewer #1: (No Response) ****** 5. Is the manuscript presented in an intelligible fashion and written in standard English? PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here. Reviewer #1: (No Response) ****** 6. Review Comments to the Author Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters) Reviewer #1: (No Response) ****** 7. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files. If you choose “no”, your identity will remain anonymous but your review may still be made public. Do you want your identity to be public for this peer review?** For information about this choice, including consent withdrawal, please see our Privacy Policy. Reviewer #1: No https://doi.org/10.1371/journal.pone.0254028.r005
Formally Accepted
22 Jun 2021 Acceptance Letter - Tatsuo Kanda, Editor PONE-D-21-14163R2 Anti-HCV antibody titer highly predicts HCV viremia in patients with hepatitis B virus dual-infection Dear Dr. Liu: I'm pleased to inform you that your manuscript has been deemed suitable for publication in PLOS ONE. Congratulations! Your manuscript is now with our production department. If your institution or institutions have a press office, please let them know about your upcoming paper now to help maximize its impact. If they'll be preparing press materials, please inform our press team within the next 48 hours. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information please contact onepress@plos.org. If we can help with anything else, please email us at plosone@plos.org. Thank you for submitting your work to PLOS ONE and supporting open access. Kind regards, PLOS ONE Editorial Office Staff on behalf of Dr. Tatsuo Kanda Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0254028.r006

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