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Monoclonal antibody-based immunohistochemistry reveals residual Taenia solium antigens in calcified granulomas from pigs with neurocysticercosis

PLOS Neglected Tropical Diseases

Dear Dr. Arroyo,

Thank you for submitting your manuscript to PLOS Neglected Tropical Diseases. After careful consideration, we feel that it has merit but does not fully meet PLOS Neglected Tropical Diseases'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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We look forward to receiving your revised manuscript.

Kind regards,

Marco Coral-Almeida, M.Sc., Ph.D.

Academic Editor

PLOS Neglected Tropical Diseases

Francesca Tamarozzi

Section Editor

PLOS Neglected Tropical Diseases

Shaden Kamhawi

co-Editor-in-Chief

PLOS Neglected Tropical Diseases

orcid.org/0000-0003-4304-636XX

Paul Brindley

co-Editor-in-Chief

PLOS Neglected Tropical Diseases

orcid.org/0000-0003-1765-0002

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Reviewers' Comments:

Reviewer's Responses to Questions

Key Review Criteria Required for Acceptance?

As you describe the new analyses required for acceptance, please consider the following:

Methods

-Are the objectives of the study clearly articulated with a clear testable hypothesis stated?

-Is the study design appropriate to address the stated objectives?

-Is the population clearly described and appropriate for the hypothesis being tested?

-Is the sample size sufficient to ensure adequate power to address the hypothesis being tested?

-Were correct statistical analysis used to support conclusions?

-Are there concerns about ethical or regulatory requirements being met?

Reviewer #1: This is a very well written manuscript, with a clearly explained objective, methodology and results section.

I only have (very) minor comments:

How were the 6 mAb selected from the 21 in house anti-Tsolium mAb? Why these 6?

Did the authors also check for inflammatation? Could they have kept sections from each block, eg the section closest to the section use for IHC?

Line 104 and 133-134: presence OF…

Reviewer #2: The methodology is well described

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Results

-Does the analysis presented match the analysis plan?

-Are the results clearly and completely presented?

-Are the figures (Tables, Images) of sufficient quality for clarity?

Reviewer #1: This is a very well written manuscript, with a clearly explained objective, methodology and results section.

Reviewer #2: the results is well described

**********

Conclusions

-Are the conclusions supported by the data presented?

-Are the limitations of analysis clearly described?

-Do the authors discuss how these data can be helpful to advance our understanding of the topic under study?

-Is public health relevance addressed?

Reviewer #1: The discussion is also well elaborated, could perhaps be shortened a bit, and includes a relevant limitation section.

Reviewer #2: The conclusion support the presented data

**********

Editorial and Data Presentation Modifications?

Use this section for editorial suggestions as well as relatively minor modifications of existing data that would enhance clarity. If the only modifications needed are minor and/or editorial, you may wish to recommend “Minor Revision” or “Accept”.

Reviewer #1: (No Response)

Reviewer #2: -

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Summary and General Comments

Use this section to provide overall comments, discuss strengths/weaknesses of the study, novelty, significance, general execution and scholarship. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. If requesting major revision, please articulate the new experiments that are needed.

Reviewer #1: This manuscript decribes the analyses of calcified cysts collected from NCC pig brains (of treated pigs), using monoclonal antibodies to identify and localise the presence of specific antigens, at different time points. Results may contribute greatly to the understanding of inflammatory processes (and seizures) around calcified cysts in human.

This is a very well written manuscript, with a clearly explained objective, methodology and results section. The discussion is also well elaborated, could perhaps be shortened a bit, and includes a relevant limitation section.

I only have (very) minor comments:

How were the 6 mAb selected from the 21 in house anti-Tsolium mAb? Why these 6?

Did the authors also check for inflammatation? Could they have kept sections from each block, eg the section closest to the section use for IHC?

Line 104 and 133-134: presence OF…

Reviewer #2: The article is well written and discussed, the methodology is acceptable, and both the methodology and results are well described. The study findings may help in understanding the neuropathology of NCC.

Some comments need to be addressed by the authors.

-Why did the authors not test for cross-reactivity using positive samples representing microbial infections?

-How does the author explain the presence of immunoreactive variabilities among the used antigens?

-The author needs to compare (with statistical significance) lesional and peri-lesional concentrations of the antigens at different time points in treated and non-treated NCC pigs to support the hypothesis of release of cyst antigens.

- Is this the first evidence of cyst antigen presence and persistence in calcified lesions and surrounding cerebral tissues? If not, state other similar evidence and comment.

-The authors claim that their findings support the hypothesis that residual parasite antigens persisting in calcified lesions and intermittently released in the perilesional brain tissue may elicit episodic inflammatory responses. However, they did not test for these hypotheses.

**********

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Reviewer #1: No

Reviewer #2: No

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Figure resubmission:--> -->-->While revising your submission, we strongly recommend that you use PLOS’s NAAS tool (https://ngplosjournals.pagemajik.ai/artanalysis) to test your figure files. NAAS can convert your figure files to the TIFF file type and meet basic requirements (such as print size, resolution), or provide you with a report on issues that do not meet our requirements and that NAAS cannot fix.-->-->

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Dear Arroyo,

We are pleased to inform you that your manuscript 'Monoclonal antibody-based immunohistochemistry reveals residual Taenia solium antigens in calcified granulomas from pigs with neurocysticercosis' has been provisionally accepted for publication in PLOS Neglected Tropical Diseases.

Before your manuscript can be formally accepted you will need to complete some formatting changes, which you will receive in a follow up email. A member of our team will be in touch with a set of requests.

Please note that your manuscript will not be scheduled for publication until you have made the required changes, so a swift response is appreciated.

IMPORTANT: The editorial review process is now complete. PLOS will only permit corrections to spelling, formatting or significant scientific errors from this point onwards. Requests for major changes, or any which affect the scientific understanding of your work, will cause delays to the publication date of your manuscript.

Should you, your institution's press office or the journal office choose to press release your paper, you will automatically be opted out of early publication. We ask that you notify us now if you or your institution is planning to press release the article. All press must be co-ordinated with PLOS.

Thank you again for supporting Open Access publishing; we are looking forward to publishing your work in PLOS Neglected Tropical Diseases.

Best regards,

Marco Coral-Almeida, M.Sc., Ph.D.

Academic Editor

PLOS Neglected Tropical Diseases

Francesca Tamarozzi

Section Editor

PLOS Neglected Tropical Diseases

Shaden Kamhawi

co-Editor-in-Chief

PLOS Neglected Tropical Diseases

orcid.org/0000-0003-4304-636XX

Paul Brindley

co-Editor-in-Chief

PLOS Neglected Tropical Diseases

orcid.org/0000-0003-1765-0002

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Reviewer's Responses to Questions

Key Review Criteria Required for Acceptance?

As you describe the new analyses required for acceptance, please consider the following:

Methods

-Are the objectives of the study clearly articulated with a clear testable hypothesis stated?

-Is the study design appropriate to address the stated objectives?

-Is the population clearly described and appropriate for the hypothesis being tested?

-Is the sample size sufficient to ensure adequate power to address the hypothesis being tested?

-Were correct statistical analysis used to support conclusions?

-Are there concerns about ethical or regulatory requirements being met?

Reviewer #1: the authors have sufficiently replied to the reviewers comments, I have no further comments.

Reviewer #2: (No Response)

**********

Results

-Does the analysis presented match the analysis plan?

-Are the results clearly and completely presented?

-Are the figures (Tables, Images) of sufficient quality for clarity?

Reviewer #1: the authors have sufficiently replied to the reviewers comments, I have no further comments.

Reviewer #2: (No Response)

**********

Conclusions

-Are the conclusions supported by the data presented?

-Are the limitations of analysis clearly described?

-Do the authors discuss how these data can be helpful to advance our understanding of the topic under study?

-Is public health relevance addressed?

Reviewer #1: the authors have sufficiently replied to the reviewers comments, I have no further comments.

Reviewer #2: (No Response)

**********

Editorial and Data Presentation Modifications?

Use this section for editorial suggestions as well as relatively minor modifications of existing data that would enhance clarity. If the only modifications needed are minor and/or editorial, you may wish to recommend “Minor Revision” or “Accept”.

Reviewer #1: (No Response)

Reviewer #2: (No Response)

**********

Summary and General Comments

Use this section to provide overall comments, discuss strengths/weaknesses of the study, novelty, significance, general execution and scholarship. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. If requesting major revision, please articulate the new experiments that are needed.

Reviewer #1: the authors have sufficiently replied to the reviewers comments, I have no further comments.

Reviewer #2: The authors addressed the reviewers' comments

**********

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

Reviewer #2: No