PONE-D-20-21835

Repeated dermal application of the common preservative methylisothiazolinone triggers local inflammation, T cell influx, and prolonged mast cell-dependent tactile sensitivity in mice

PLOS ONE

Dear Dr. Chatterjea,

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Praveen Thumbikat

Academic Editor

PLOS ONE

Additional Editor Comments:

This manuscript by Kline et al., expands on prior work by this group in models of vulvodynia. This particular manuscript follows familiar ground in describing increased perigenital skin sensitivity and a role for mast cells in the methylisothiazolinone induced mouse model. New and noteworthy experiments in this study include the characterization of the immune compartment including T cells, neutrophils and soluble cytokines at various times in the model. In addition, the authors use imatinib as a new agent to inhibit mast cell signaling and show some efficacy in reducing symptoms in the mouse model. While the study itself is well designed and has adequate controls, a number of concerns do remain that would need to be addressed. They are listed below in no particular order.

1. Sections of the introduction are extremely similar/verbatim to a recent publication from the group.

2. In the mechanical hypersensitivity assays shown - all the results are shown after normalization (percent decrease etc). It is difficult to appreciate the relevance of the decreases without seeing the absolute values - perhaps a table can be shown in supplementary data to complement the normalized data shown in figures.

3. The increases in NGF and cadm1 in the spinal cord is interesting, but its direct relevance to the current study is limited and therefore its inclusion in the supplementary data needs to be reconsidered.

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Reviewers' comments:

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Comments to the Author

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

Reviewer #1: Yes

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Reviewer #1: Yes

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4. Is the manuscript presented in an intelligible fashion and written in standard English?

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Reviewer #1: 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: This manuscript by Kline J., et al., aims to study the effects of exposure to a common household biocide preservative – methylisothiazolinone (MI) on the labiar skin in the development of genital pain and associated inflammatory changes. In this study, the authors use repeated MI challenges to sensitize the skin and trigger an inflammatory reaction as well as ano-genital sensitivity. They observe increases in inflammatory cytokines, as well as increases in the immune cell infiltrates; including neutrophils, eosinophils, mast cells, and T cells upon MI challenges, which causes a dramatic and persistent increase in ano-genital sensitivity. They conclude the manuscript showing that a therapeutic administration of imatinib causes an alleviation of tactile ano-genital sensitivity, suggesting a possible pivotal role played by mast cells and opens up targets for therapeutic approaches.

The manuscript, as it stands, is however not novel as most of the results and observations reported in the Figures (1, 3, and 4) have been published in their previous publication as cited in the manuscript as Arriaga-Gomez E., et al. [14]. The previously published data showed increase in mast cells, ano-genital sensitivity as well as increased pro-inflammatory cytokines IL6 and IFN-gamma. The only difference between the previous manuscript seems to be the mode of administration of MI, which leads to alteration in the persistence of the ano-genital sensitivity. Furthermore, the previously published article already addressed the issue of the role played by mast cells in driving ano-genital sensitivity using THC as a therapeutic approach.

Major Weaknesses:

1. This study lacks novelty and most of the results have been already previously published by this group.

2. CD44 +CD25+ CD4 T cells as well CD44+ CD8 T cells do not represent T cell subtypes but rather activation states of the T cells. It is important to delineate between T cell subtypes and activation state.

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Reviewer #1: No

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Repeated dermal application of the common preservative methylisothiazolinone triggers local inflammation, T cell influx, and prolonged mast cell-dependent tactile sensitivity in mice

PONE-D-20-21835R1

Dear Dr. Chatterjea,

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.

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Kind regards,

Praveen Thumbikat

Section 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: 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

**********

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

**********

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

**********

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 have addressed all the critics raised. The manuscript in the current form is well written and concise.

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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.

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Reviewer #1: No