Vaginal microbiota and personal risk factors associated with HPV status conversion—A new approach to reduce the risk of cervical cancer?

Zhongzhou Yang; Ye Zhang; Araceli Stubbe-Espejel; Yumei Zhao; Mengping Liu; Jianjun Li; Yanping Zhao; Guoqing Tong; Na Liu; Le Qi; Andrew Hutchins; Songqing Lin; Yantao Li

doi:10.1371/journal.pone.0270521

Peer Review History

Original SubmissionDecember 14, 2021
2 Mar 2022 Decision Letter - Guangming Zhong, Editor PONE-D-21-39382Vaginal microbiota and personal risk factors associated with HPV status conversion – a new approach to reduce the risk of cervical cancer?PLOS ONE Dear Dr. Yang, 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 Apr 16 2022 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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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: Yes ******** 2. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: Yes ****** 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). 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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: Human papillomavirus (HPV), being considered as a sexually transmitted pathogen, which is responsible for over 90% of cervical cancer cases, poses a severe threat to woman’s reproductive health. Though the factors affecting HPV-persistence are not fully understood, emerging data suggests that there exists an association between cervical HPV infections and the vaginal microbiota. This study demonstrated that several factors significantly associated with persistent HPV infection, including age, salary, history of reproductive tract infection, and the total number of sexual partners and in vaginal microbiota, Lactobacillus gasseri, Streptococcus agalactiae, and Timona prevotella bacteria may be associated with HPV clearance. Besides, by using the eHealth platform which is a user-friendly mobile application program, it is more cost-efficient than any other kind of management for researchers to pay full attention to every participant in a health-related screening program. 1. Major Line 101 to line 104: We found the participant was requested to abstain from vaginal intercourse 24 hours before sampling, to wait for at least three days after menstrual blood was cleared and to avoid using vaginal douches and any vaginally administered medical treatments. But at different stages of the menstrual cycle, such as follicular phase and the luteal phase, physiological changes could have an impact on vaginal flora growth, colonization, and community structure. It is suggested to collect sample of vaginal mucus at the same stage of the menstrual cycle or indicate what stage of the menstrual cycle the collected samples come from. 2. Minor 1) Line 105 to line 106: Sample of vaginal mucus was collected by inserting a swab into the vagina, which was then stirred/placed inside a special tube with a preservative solution at room temperature until the pick-up was arranged. Is this preservative solution a DNA protection solution? 2) Line 277: “HPV-negative to HPV-negative” would be “HPV positive to HPV negative”. ****** 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 [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.0270521.r001
Revision 1
26 Mar 2022 Author Response Dear Reviewer: Sincerely thanks for your three comments. They are responding in below: 1. Major Line 101 to line 104: We found the participant was requested to abstain from vaginal intercourse 24 hours before sampling, to wait for at least three days after menstrual blood was cleared and to avoid using vaginal douches and any vaginally administered medical treatments. But at different stages of the menstrual cycle, such as follicular phase and the luteal phase, physiological changes could have an impact on vaginal flora growth, colonization, and community structure. It is suggested to collect sample of vaginal mucus at the same stage of the menstrual cycle or indicate what stage of the menstrual cycle the collected samples come from. Thanks for your questions. We collected samples after 3 days beyond the menses. It can be both follicular and luteal phase because of three reasons with four references as in manuscript. 1.1, According to Stephanie et al, vaginal microbial diversity, as measured using the Shannon index, increased during menses blood (P < 0.001), while Lactobacillus abundances decreased (P = 0.01). Hence, we did not collect samples during menses. 1.2, Based on Bonnie et al, the overall vaginal microbiome of most women remained relatively stable throughout the menstrual cycle, with little variation in diversity and only modest fluctuations in species richness. That is to say, there is little variation between follicular and luteal phase. 1.3, The reason why it was three days accounted by Pawel et al. In their article, Figure S6 showed Shannon diversity indices over the menstrual time. Based on the figure, the diversity index decreased to one and less after three days in the end of menses. 1.4, Although samples obtained during a menstrual period should be valid, most women would prefer to obtain the sample at a time other than during their menstrual flow based on Jerome et al. Therefore, we summarized the descriptions for: to wait for at least three days after menstrual blood was cleared. 2. Minor 1) Line 105 to line 106: Sample of vaginal mucus was collected by inserting a swab into the vagina, which was then stirred/placed inside a special tube with a preservative solution at room temperature until the pick-up was arranged. Is this preservative solution a DNA protection solution? Thanks for your questions. Exactly, this solution is a DNA protection solution through N-octylpyridinium bromide (NOPB) published in Han-Microbiome-2018 from my institution. Furthermore, the NOPB was also acknowledged by article Qian-Chin Med J (Engl)-2020. Two articles are shown in manuscripts 2) Line 277: “HPV-negative to HPV-negative” would be “HPV positive to HPV negative”. Thanks for your suggestions. It was updated in accordingly. Dear Editor: With regard to your three additional requirements, they were responded in below: 1. Please ensure that your manuscript meets PLOS ONE's style requirements, including those for file naming. Responses: To easy review for you, files have been updated carefully to ensure meets PLOS ONE's style requirements after reading your suggested documents. 2. Stating the financial disclosure: Responses: This study was supported by funding from the Shenzhen Innovation Committee of Science and Technology (ZDSYS20200811144002008). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. 3. Stating the competing interests section: Responses: The authors have declared that no competing interests exist. Jacri Attachments Attachment Submitted filename: Response reviewers comments.docx https://doi.org/10.1371/journal.pone.0270521.r002
12 Jun 2022 Decision Letter - Guangming Zhong, Editor Vaginal microbiota and personal risk factors associated with HPV status conversion – a new approach to reduce the risk of cervical cancer? PONE-D-21-39382R1 Dear Dr. Yang, 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, Guangming Zhong Academic Editor PLOS ONE Additional Editor Comments (optional): Reviewers' comments: https://doi.org/10.1371/journal.pone.0270521.r003
Formally Accepted
29 Jul 2022 Acceptance Letter - Guangming Zhong, Editor PONE-D-21-39382R1 Vaginal microbiota and personal risk factors associated with HPV status conversion – a new approach to reduce the risk of cervical cancer? Dear Dr. Yang: 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. Guangming Zhong Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0270521.r004

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