Aberrant HLA-DR expression in the conjunctival epithelium after autologous serum treatment in patients with graft-versus-host disease or Sjögren’s syndrome

Katerina Jirsova; Petra Seidler Stangova; Michalis Palos; Gabriela Mahelkova; Sarka Kalasova; Ivana Rybickova; Tor Paaske Utheim; Viera Vesela

doi:10.1371/journal.pone.0231473

Peer Review History

Original SubmissionSeptember 26, 2019
24 Dec 2019 Decision Letter - Alexander V. Ljubimov, Editor PONE-D-19-27094 Aberrant HLA-DR expression in the conjunctival epithelium after autologous serum treatment in patients with graft-versus-host disease or Sjögren’s syndrome PLOS ONE Dear Assoc. Prof., Jirsova, 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 authors performed an observational study of subjects with dry eye either secondary to oGVHD or primary Sjogren's syndrome to correlate clinical symptoms with impression cytology of the conjunctiva and to determine the number of HLADR+ cells before and after treatment with autologous serum eye drops. The reviewers found some problems with the paper that the authors need to address. 1. It is noted that there are many confounders in study - sex, previous treatment, reliability of counting + HLADR cells from IC stained filters. These need to be evaluated. 2. It is not clear if the readers of IC were " blinded" to pre vs. post AS treatment when evaluating samples. We would appreciate receiving your revised manuscript by Feb 07 2020 11:59PM. When you are 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. If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. To enhance the reproducibility of your results, we recommend that if applicable you deposit your laboratory protocols in protocols.io, where a protocol can be assigned its own identifier (DOI) such that it can be cited independently in the future. 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We look forward to receiving your revised manuscript. Kind regards, Alexander V. Ljubimov, Ph.D. Academic Editor PLOS ONE Journal Requirements: 1. When submitting your revision, we need you to address these additional requirements. Please ensure that your manuscript meets PLOS ONE's style requirements, including those for file naming. The PLOS ONE style templates can be found at http://www.journals.plos.org/plosone/s/file?id=wjVg/PLOSOne_formatting_sample_main_body.pdf and http://www.journals.plos.org/plosone/s/file?id=ba62/PLOSOne_formatting_sample_title_authors_affiliations.pdf 2. Please provide additional details regarding participant consent. In the Methods section, please ensure that you have specified what type of consent you obtained (for instance, written or verbal) and whether the ethics committee approved this consent procedure. If verbal consent was obtained please state why it was not possible to obtain written consent and how verbal consent was recorded. If your study included minors, state whether you obtained consent from parents or guardians. 3. Please ensure that you have provided a limitations section in your manuscript. This section should discuss the absence of a control/placebo group in your study. We noticed you have some minor occurrence of overlapping text with the following previous work, which needs to be addressed: https://doi.org/10.3109/02713683.2013.824987 In your revision ensure you cite all your sources (including your own works), and quote or rephrase any duplicated text outside the methods section. Further consideration is dependent on these concerns being addressed. Additional Editor Comments: The authors performed an observational study of subjects with dry eye either secondary to oGVHD or primary Sjogren's syndrome to correlate clinical symptoms with impression cytology of the conjunctiva and to determine the number of HLADR+ cells before and after treatment with autologous serum eye drops. The reviewers found some problems with the paper that the authors need to address. 1. It is noted that there are many confounders in study - sex, previous treatment, reliability of counting + HLADR cells from IC stained filters. These need to be evaluated. 2. It is not clear if the readers of IC were " blinded" to pre vs. post AS treatment when evaluating samples. [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 ******** 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). 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 ****** 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 ****** 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: Obervational study of subjects with dry eyes either secondary to oGVHD or primary Sjogren's syndrome to correlated clinical symptoms with impression cytology of the conjunctiva to determine number of HLADR + cells before and after treatment with autologous serum eye drops. Many confounders in study- sex , previous treatment, reliability of counting + HLAdr cells from IC stained filters. Also not clear if readers of IC were " blinded" to pre vs post AS treatment when evaluating samples. Study is interesting, but given potential confounders may be difficult to reach a conclusion. Most interesting part of study is the use of aberrant HLADR staining for positivity of conj. epithelial cells- this topic could be explored further ****** 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 to be viewed.] 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 us at figures@plos.org. Please note that Supporting Information files do not need this step. https://doi.org/10.1371/journal.pone.0231473.r001
Revision 1
28 Feb 2020 Author Response February 4, 2020 Alexander V. Ljubimov, Ph.D. Academic Editor PLOS ONE Dear Dr. Ljubimov, We thank the reviewers for their helpful comments, and we have addressed the comments from the referees as follows: PONE-D-19-27094 Response to Reviewers 1 The authors performed an observational study of subjects with dry eye either secondary to o GVHD or primary Sjogren's syndrome to correlate clinical symptoms with impression cytology of the conjunctiva and to determine the number of HLADR+ cells before and after treatment with autologous serum eye drops. The reviewers found some problems with the paper that the authors need to address. It is noted that there are many confounders in study - sex, previous treatment, reliability of counting + HLADR cells from IC stained filters. These need to be evaluated. Potential confounder factors has been analyzed where possible. No statistical difference was found in the age of patients with GVHD and SS respectively (p=0,94). See Statistical analysis and Results sections in the manuscript. Due to the fact that group of SS involves just females, and GVHD patients included both males and females, statistical difference of HLA-Dr positivity was calculated separately for GVHD males and GVHD females. No significant difference was found. This information was included in the manuscript. The paragraph regarding treatment of patients (Material and Methods section) was rewritten separately for GVHD and SS patients respectively ((“Sixteen patients were under the treatment with preservative-free artificial tear eye drops (nine from GVHD and seven from SS group respectively), and eight patients with artificial tears containing oxychloro complex (Purite) or Polyquad preservatives (four patients per each group). Seven patients, all from the GvHD group, were on systemic immunosuppressive therapy and antiviral therapy. In the SS group, two patients were receiving systemic corticosteroids.”)). The therapy of dry eye disease is highly individualized, the broad spectrum of medical treatment does not allow statistical analysis. It should be, however, noted that therapy regimen was not changed for any patient involved in the study for at least two months prior to the application of AS and during the follow-up period, which we consider to be an important factor. The sentence ”The HLA-DR-positive cells were clearly discernible from negative cells.” was added to Material and Methods section. 2. It is not clear if the readers of IC were " blinded" to pre vs. post AS treatment when evaluating samples. Images were evaluated by two independent investigators blinded to the experimental conditions (patient identity, before and after AS specimens). Following sentence was added to the text. “The density of HLA-DR-positive epithelial cells and Langerhans cells was assessed independently by two researchers (KJ, VV) blinded to the experimental conditions using a NIS Elements image analysis system (Laboratory Imaging, Prague, Czech Republic).” Yours sincerely, Katerina Jirsova Assoc. Prof. Katerina Jirsova, Ph.D. Laboratory of the Biology and Pathology of the Eye Institute of Biology and Medical Genetics, First Faculty of Medicine, Charles University and General University Hospital in Prague, Albertov 4, 128 00 Prague 2, Czech Republic Tel: 00420 224 968 006, E-mail: katerina.jirsova@lf1.cuni.cz Attachments Attachment Submitted filename: Response to Reviewers.docx https://doi.org/10.1371/journal.pone.0231473.r002
25 Mar 2020 Decision Letter - Alexander V. Ljubimov, Editor Aberrant HLA-DR expression in the conjunctival epithelium after autologous serum treatment in patients with graft-versus-host disease or Sjögren’s syndrome PONE-D-19-27094R1 Dear Dr. Jirsova, We are pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it complies with all outstanding technical requirements. Within one week, you will receive an e-mail containing information on the amendments required prior to publication. When all required modifications have been addressed, you will receive a formal acceptance letter and your manuscript will proceed to our production department and be scheduled for publication. Shortly after the formal acceptance letter is sent, an invoice for payment will follow. To ensure an efficient production and billing process, please log into Editorial Manager at https://www.editorialmanager.com/pone/, click the "Update My Information" link at the top of the page, and update your user information. 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 enable them to help maximize its impact. If they will be preparing press materials for this manuscript, you must inform our press team as soon as possible and 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. With kind regards, Alexander V. Ljubimov, Ph.D. Academic Editor PLOS ONE Additional Editor Comments (optional): The authors adequately replied to the critique. Reviewers' comments: https://doi.org/10.1371/journal.pone.0231473.r003
Formally Accepted
9 Apr 2020 Acceptance Letter - Alexander V. Ljubimov, Editor PONE-D-19-27094R1 Aberrant HLA-DR expression in the conjunctival epithelium after autologous serum treatment in patients with graft-versus-host disease or Sjögren’s syndrome Dear Dr. Jirsova: I am 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 notify them about your upcoming paper at this point, to enable them to help maximize its impact. If they will be preparing press materials for this manuscript, 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. For any other questions or concerns, please email plosone@plos.org. Thank you for submitting your work to PLOS ONE. With kind regards, PLOS ONE Editorial Office Staff on behalf of Dr. Alexander V. Ljubimov Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0231473.r004

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