The diagnostic value of decreased levels of inflammatory markers for the state of hepatitis C virus (HCV) infection

Lu Han; Lei Bi; Xinpeng Li

doi:10.1371/journal.pone.0332105

Peer Review History

Original SubmissionApril 30, 2025
28 May 2025 Decision Letter - Maemu Petronella Gededzha, Editor Dear Dr. Li, 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. The document contains spelling errors and requires grammatical editing. For example, in Table 1, the word 'unknow' should be corrected. A thorough revision is recommended. The abbreviation ‘COVID-19’ is inconsistently written as both ‘COVID-19’ and ' covid-19' in different parts of the text. Please standardize its formatting There are no page and line numbers used in the document- the authors did not follow PLOS instructions and makes it difficult to refer. 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The PLOS ONE style templates can be found at https://journals.plos.org/plosone/s/file?id=wjVg/PLOSOne_formatting_sample_main_body.pdf and https://journals.plos.org/plosone/s/file?id=ba62/PLOSOne_formatting_sample_title_authors_affiliations.pdf 2. 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 If you have cited papers that have been retracted, please include the rationale for doing so in the manuscript text, or remove these references and replace them with relevant current references.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 If you need to cite a retracted article, indicate the article’s retracted status in the References list and also include a citation and full reference for the retraction notice. Additional Editor Comments: The document contains spelling errors and requires grammatical editing. For example, in Table 1, the word 'unknow' should be corrected. A thorough revision is recommended. The abbreviation ‘COVID-19’ is inconsistently written as both ‘COVID-19’ and ' covid-19' in different parts of the text. Please standardize its formatting There are no page and line numbers used in the document- the authors did not follow PLOS instructions and makes it difficult to refer [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? Reviewer #1: Yes Reviewer #2: Partly ******** 2. Has the statistical analysis been performed appropriately and rigorously? -->?> Reviewer #1: Yes Reviewer #2: Yes ****** 3. Have the authors made all data underlying the findings in their manuscript fully available??> The PLOS Data policy Reviewer #1: Yes Reviewer #2: Yes ****** 4. Is the manuscript presented in an intelligible fashion and written in standard English??> Reviewer #1: Yes Reviewer #2: Yes ****** Reviewer #1: 2. Methods 2.1 HCV Diagnosis There is a mention of molecular assays used by NHANES but not for anti-HCV (antibody-based) and the anti-HCV was used to categorize participants into anti-HCV pos or negative. 2.3 Study Population Figure 1 and the paragraph above (that explains exclusion and sample size) on page 6 and 7, the numbers do not correlate – the figure states that “14007” participants enrolled (please correct grammar of enrolled (‘entolled’) in the last box of the figure) and the first sentence in pg 7 says 17 526 were enrolled. Please revise also the exclusion in the text and figure 1 for instance “blood metals” no where in the text is mentioned as exclusion criteria, as well as ≥18 years old on the excluded part-revise this as this impacts on age stratification. Revise numbers in the excluded in figure 1 in conjunction with those in the text above the figure 1. 2.5 Statistical Analysis Page 8 12th row-…”predict sensitivity and specialist”.. it seems there is grammar error, was it not meant to be “specificity” instead of “specialist” 3. Results 3.1 Characteristics of participants (pg 8-9) Results are based on total participants of 17 526, which is not in agreement with figure 1 which shows exclusion and enrollment of participants coming up to 14 007. Kindly address 2.3 of Methods to correctly reflect analyzed population number. 4. Discussion Page 21 14th row- Please review and revise the sentence, grammar seems to be incomplete, sentence has no significance meaning as it written currently Reviewer #2: The manuscript is well written, with detailed statistics however it would have been better had the authors detailed cytokines aligned to the ratios it would have given depth and a clearer understanding of the inflammatory mechanism in the study. The inability of the ratios to have any diagnostic value for those below 30 is a major limitation to recommend the use of the ratios for diagnostic purposes. The OR are also not well represented and are difficult to interpret on current form. Also, although PLR, PMR and NMR show a negative association with HCV, this is not properly ventilated in the discussion. If it is possible to address these minor comments, this is a well written and informative paper on HCV diagnosis. ****** 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? 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Revision 1
22 Jul 2025 Author Response Manuscript ID: PONE-D-25-23527 Manuscript Title: The Diagnostic Value of Decreased Levels of Inflammatory Markers for the State of Hepatitis C Virus (HCV) Infection Authors: Lu Han, Lei Bi, Xinpeng Li* The authors appreciate the professional comments from the editors and reviewers. According to the comments, we have made extensive modifications to our manuscript to make our results convincing. All spelling errors have been corrected as the suggestions. The reference format has been revised to the “Vancouver” style as required by submission guidelines. Our point-by-point responses were presented as follows: Response to Reviewer 1: 2.1 HCV Diagnosis There is a mention of molecular assays used by NHANES but not for anti-HCV (antibody-based) and the anti-HCV was used to categorize participants into anti-HCV pos or negative. The author’s answer: We have added specific details regarding HCV antibody testing in Methods 2.2, lines 104-116. The content is as follows: HCV antibody was tested based on the VITROS Anti-HCV Reagent Pack and VITROS Immunodiagnostic Products Anti-HCV Calibrator on the VITROS ECi/ECiQ Immunodiagnostic Systems, the VITROS 3600 Immunodiagnostic System and the VITROS 5600 Integrated System. This involves a two-stage reaction. In the first stage, HCV antibody present in the sample binds with HCV recombinant antigens coated on the wells. Unbound sample is removed by washing. In the second stage, horseradish peroxidase (HRP)-labeled antibody conjugate (mouse monoclonal anti-human IgG) binds to any human IgG captured on the well in the first stage. Unbound conjugate is removed by washing. The system signal intensity is measured by chemiluminescence, and the signal intensity is directly proportional to the anti-HCV level in the sample. 2.3 Study Population Figure 1 and the paragraph above (that explains exclusion and sample size) on page 6 and 7, the numbers do not correlate – the figure states that “14007” participants enrolled (please correct grammar of enrolled (‘entolled’) in the last box of the figure) and the first sentence in pg 7 says 17 526 were enrolled. Please revise also the exclusion in the text and figure 1 for instance “blood metals” no where in the text is mentioned as exclusion criteria, as well as ≥18 years old on the excluded part-revise this as this impacts on age stratification. Revise numbers in the excluded in figure 1 in conjunction with those in the text above the figure 1 The author’s answer: Thank you for your careful correction. We previously performed data adjustment for inclusion and exclusion, but we overlooked updating the data in the article's sections. We have now thoroughly reviewed and verified everything. The total number of participants from 2007 to 2018 is 59,842, and the final number of participants included in this study is 17,526. The corresponding spelling errors have also been corrected. Please refer to Figure 1 for details, with further information provided in lines 137-143. 2.5 Statistical Analysis Page 8 12th row-…”predict sensitivity and specialist”.. it seems there is grammar error, was it not meant to be “specificity” instead of “specialist” The author’s answer: The grammatical error has been corrected to "specificity," as detailed in line 172. 3. Results 3.1 Characteristics of participants (pg 8-9) Results are based on total participants of 17 526, which is not in agreement with figure 1 which shows exclusion and enrollment of participants coming up to 14 007. Kindly address 2.3 of Methods to correctly reflect analyzed population number. The author’s answer: This issue has been corrected, and the final number of participants included in this study is 17,526. 4.Discussion Page 21 14th row- Please review and revise the sentence, grammar seems to be incomplete, sentence has no significance meaning as it written currently. The author’s answer: The sentence indeed contained a grammatical error and has now been revised to: "As a cross-sectional analysis based on NHANES data, our study relies on single-time sampling and survey data without access to longitudinal follow-up. Therefore, it cannot adequately address the potential value of continuous monitoring of these inflammatory markers." Please refer to lines 338-342 for further details. Response to Reviewer 2: 1.The manuscript is well written, with detailed statistics however it would have been better had the authors detailed cytokines aligned to the ratios it would have given depth and a clearer understanding of the inflammatory mechanism in the study. The author’s answer: We have reviewed some literature to clarify the relationship between the ratio, HCV, and cytokines in the discussion. Relevant content has been added to the discussion section in lines 325-336. Research indicates that approximately half of patients with community-acquired acute hepatitis C can spontaneously clear the virus, and significantly higher levels of IL-1β, IL-1RA, IL-6, IFN-γ, and FGF-2 were detected in the serum of patients achieving self-limited infection. Microbiota-modulated cytokines such as IL-1β, IL-18, and IL-22 can promote antiviral defense mechanisms by regulating adaptive immune responses. However, when the symbiotic state shifts toward pathology, these same antiviral cytokines paradoxically drive more severe disease progression to cirrhosis and hepatocellular carcinoma. Routine cytokine profiling remains impractical for large-scale screening or asymptomatic subclinical populations. Therefore, Blood-cell-based inflammatory biomarkers — some studies showed were closely associated with cytokine networks — may provide cost-effective insights into antiviral potential and chronic infection progression. 2. The inability of the ratios to have any diagnostic value for those below 30 is a major limitation to recommend the use of the ratios for diagnostic purposes. The author’s answer: Regarding the lack of diagnostic value of the ratio in the population under 30 years old in this study, we have added this point in the last paragraph of the discussion section, lines 342-346. Based on the findings of this study, Blood-cell-based inflammatory biomarkers demonstrate no clinically significant correlation with HCV progression in individuals under 30 years of age. Consequently, there is insufficient evidence to support their utility for monitoring HCV progression in this demographic, necessitating continued reliance on alternative diagnostic modalities. 3. The OR are also not well represented and are difficult to interpret on current form. Also, although PLR, PMR and NMR show a negative association with HCV, this is not properly ventilated in the discussion. The author’s answer: This negative association is primarily caused by thrombocytopenia resulting from hepatitis virus infection. We have added an explanation in the discussion section, lines 309-315. Hepatitis virus infection has a long course of disease, and long-term hepatitis virus infection can lead to hypersplenism and portal hypertension leading to thrombocytopenia, while the virus activates lymphocytes. In the case of respiratory virus infection, because of its short course of illness, usually 5-7 days, during this process, platelets can be activated as inflammatory factors, and influenza virus infection can also lead to lymphocyte apoptosis and less circulating lymphocyte. Attachments Attachment Submitted filename: Response to Reviewers.docx https://doi.org/10.1371/journal.pone.0332105.r002
27 Aug 2025 Decision Letter - Maemu Petronella Gededzha, Editor The Diagnostic Value of Decreased Levels of Inflammatory Markers for the State of Hepatitis C Virus (HCV) Infection PONE-D-25-23527R1 Dear Dr. Xinpeng Li, 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 will be generated when your article is formally accepted. Please note, if your institution has a publishing partnership with PLOS and your article meets the relevant criteria, all or part of your publication costs will be covered. Please make sure your user information is up-to-date by logging into Editorial Manager at Editorial Manager® and clicking the ‘Update My Information' link at the top of the page. For questions related to billing, please contact billing support . 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, Maemu Petronella Gededzha, Ph.D Academic Editor PLOS ONE Additional Editor Comments (optional): Reviewers' comments: Reviewer's Responses to Questions Comments to the Author Reviewer #1: All comments have been addressed Reviewer #2: All comments have been addressed ******** 2. Is the manuscript technically sound, and do the data support the conclusions??> Reviewer #1: Partly Reviewer #2: Yes ****** 3. Has the statistical analysis been performed appropriately and rigorously? -->?> Reviewer #1: Yes Reviewer #2: Yes ****** 4. Have the authors made all data underlying the findings in their manuscript fully available??> The PLOS Data policy Reviewer #1: Yes Reviewer #2: Yes ****** 5. Is the manuscript presented in an intelligible fashion and written in standard English??> Reviewer #1: Yes Reviewer #2: Yes ****** Reviewer #1: (No Response) Reviewer #2: The authors have addressed all the queries raised in the first review, I am happy to recommend this paper for publication. ****** 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 ******** https://doi.org/10.1371/journal.pone.0332105.r003
Formally Accepted
Acceptance Letter - Maemu Petronella Gededzha, Editor PONE-D-25-23527R1 PLOS ONE Dear Dr. Li, I'm pleased to inform you that your manuscript has been deemed suitable for publication in PLOS ONE. Congratulations! Your manuscript is now being handed over to our production team. At this stage, our production department will prepare your paper for publication. This includes ensuring the following: * All references, tables, and figures are properly cited * All relevant supporting information is included in the manuscript submission, * There are no issues that prevent the paper from being properly typeset You will receive further instructions from the production team, including instructions on how to review your proof when it is ready. Please keep in mind that we are working through a large volume of accepted articles, so please give us a few days to review your paper and let you know the next and final steps. Lastly, 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. You will receive an invoice from PLOS for your publication fee after your manuscript has reached the completed accept phase. If you receive an email requesting payment before acceptance or for any other service, this may be a phishing scheme. Learn how to identify phishing emails and protect your accounts at https://explore.plos.org/phishing. If we can help with anything else, please email us at customercare@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. Maemu Petronella Gededzha Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0332105.r004

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