PONE-D-19-21637

Immunopathogenesis of Canine Chronic Ulcerative Stomatitis

PLOS ONE

Dear Dr Anderson

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.

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Many thanks for submitting your manuscript to PLOS One

After difficulties in getting appropriate reviewers, two reviewers were found

One recommended a rejection and the other recommended to accept

I have therefore decided on a major revision. If you feel that you can address reviewer one's comments then please do so and resubmit

I will send the resubmitted manuscript to the same reviewer.

I wish you good luck with your revisions

Thanks

Simon

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We would appreciate receiving your revised manuscript by Dec 21 2019 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.

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We look forward to receiving your revised manuscript.

Kind regards,

Simon Russell Clegg, PhD

Academic Editor

PLOS ONE

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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: Yes

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2. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: Yes

Reviewer #2: Yes

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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: No

Reviewer #2: Yes

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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

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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: The authors sought to investigate some leukocyte populations by examining lesions from dogs with chronic ulcerative stomatitis. Nonetheless, there are many concerns described below:

1. The introduction is too long, and just in this section the authors cited 45 articles. I strongly suggest the authors to be concice.

2. Line 241. Why the authors used HE or toluidine blue? Why the authors did not standardize the dye used to stain mucosa tissue?

3. One of my major concerns are the antibodies used in immunohistochemistry. What is the specificify of the antibodies used for dogs? The cross-reactivity of all antibodies used must be proven (e.g. anti-CD20, anti-Mum1 and anti-CD204)? How the authors titrate the antibodies? All antibodies used to identify leukocyte populations are monoclonal antibodies (e.g. anti-CD20)? This information should be clearly stated. Why the authors did not include a negative control in immunohistochemistry assay? The controls of immunohistochemistry should be carefully conducted (please, for more details see: Hewitt et al., J Histochem Cytochem. 2014). How the authors select their antibodies? Why the authors used CD20 to identify B cells ("CD20 is expressed primarily on B cells but has also been detected on both normal and neoplastic T cells" available at https://www.thermofisher.com/order/catalog/product/RB-9013-P0#/RB-9013-P0? Why the authors used anti-CD204 (a macrophage scavenger receptor) antibody to identify macrophages?

4. Please, provide clearly the information about the monoclonal antibodies used to identify CD4+ and CD8+ cells. It is also a pity that CD4 and CD8 antibodies were not used together. Unfortunately, the importance of T CD4 CD8 double negative (e.g Fisher et al., Blood, 2005; D'Acquisto, Biochemical Phamarcology, 2011) and even positive (e.g. Clénet et al., Sci. Rep., 2019) cells was neglected in the present study. Furthermore, it would be great if the macrophage subsets has been considered.

5. The cross-reactivity of anti-human IL17 antibody must be determined/shown. The specifications of all antibodies used should be clearly stated (if possible, include the catalog number).

6. It is a pity that the authors did not quantify IL-17 by qPCR in tissue, and the producing cells is not clearly identified.

7. Line 389-390. The PloS One rules recommended that al data should be clearly showed.

8. The conclusion section should be profoundly revised.

Reviewer #2: This was a well designed and rigorously performed analysis of the pathological processes in this frustrating disease of the canine oral cavity. It is good to see mounting evidence of the likely immune-mediated pathway, in order that further research can hopefully lead to better methods of control or cure.

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Reviewer #1: No

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Immunopathogenesis of Canine Chronic Ulcerative Stomatitis

PONE-D-19-21637R1

Dear Dr. Anderson

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.

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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,

Simon Russell Clegg, PhD

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

Many thanks for your re submission to PLOS One

The manuscript was reviewed by the same reviewer as last time, and they have recommended acceptance of the manuscript

I wish to congratulate you on your research, and wish you all the best for the future

Many thanks

Simon

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: All comments have been addressed

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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: Yes

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3. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: 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: Yes

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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: Yes

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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: The authors performed a great job providing valuable information for the scientific community on the immunopathogenesis of canine chronic ulcerative stomatitis, and all my previous concerns were properly addressed.

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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: Yes: Fernando Nogueira Souza