Transfer Alert

This paper was transferred from another journal. As a result, its full editorial history (including decision letters, peer reviews and author responses) may not be present.

Dear Mrs. Hoving,

Thank you very much for submitting your manuscript "The pathogenesis of experimental Emergomycosis in mice." for consideration at PLOS Neglected Tropical Diseases. As with all papers reviewed by the journal, your manuscript was reviewed by members of the editorial board and by several independent reviewers. The reviewers appreciated the attention to an important topic. Based on the reviews, we are likely to accept this manuscript for publication, providing that you modify the manuscript according to the review recommendations.

Please prepare and submit your revised manuscript within 30 days. If you anticipate any delay, please let us know the expected resubmission date by replying to this email.

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[1] A letter containing a detailed list of your responses to all review comments, and a description of the changes you have made in the manuscript.

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[2] Two versions of the revised manuscript: one with either highlights or tracked changes denoting where the text has been changed; the other a clean version (uploaded as the manuscript file).

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Thank you again for your submission to our journal. We hope that our editorial process has been constructive so far, and we welcome your feedback at any time. Please don't hesitate to contact us if you have any questions or comments.

Sincerely,

Ahmed Hassan Fahal, FRCS, FRCSI, FRCSG, MS, MD, FRCP(London)

Academic Editor

PLOS Neglected Tropical Diseases

Marcio Rodrigues

Section Editor

PLOS Neglected Tropical Diseases

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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 study was well designed and the authors have managed to establish a novel murine model of E. africanus disease providing critical insights into the host immune components required for eliminating the infection. No concerns about the ethical or regulatory requirement met.

The methods were suitable to reach a sensible and measurable outcomes.

Reviewer #2: -Are the objectives of the study clearly articulated with a clear testable hypothesis stated? Yes

-Is the study design appropriate to address the stated objectives? Not entirely

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

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

-Were correct statistical analysis used to support conclusions? Yes

-Are there concerns about ethical or regulatory requirements being met? No concerns

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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: The analysis of data displayed matches the analysis plan. The results are extensive but clear and one cytokine (IL 10) results needs to be displayed in the abstract because it is key cytokine antagonising interferon gamma. Good quality figures and images.

Reviewer #2: -Does the analysis presented match the analysis plan? Yes

-Are the results clearly and completely presented? Yes

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

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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: Extracted from the results.

Reviewer #2: -Are the conclusions supported by the data presented? Not entirely

-Are the limitations of analysis clearly described? Yes

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

-Is public health relevance addressed? Yes

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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: IL 10 results can be presented in the abstract as key cytokine when addressing IFN-gamma and immunopathogenesis.

Reviewer #2: There is a spelling error on line 310.

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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: Well designed and conducted study. Acceptable for publication.

Reviewer #2: This is a study to establish a mouse model of pulmonary Emergomyces africanus infection that is observed in some HIV patients with advanced disease. To further understand the biology of E. Africanus infections, the authors utilize Rag 1 deficient mice.

The overall strengths of this study were the novelty of the model, the use of different mouse models with Th1 and Th2- biased responses and some mechanistic insights from the Rag1 deficient mice. The data was clearly presented with impactful imagery. Indeed, the authors were able to establish infection in various tissues that persisted for 2 or more weeks, therefore this is a feasible experimental model.

However, there were weaknesses in the study: the association with advanced HIV in the experimental model was not shown, therefore, I think that associations with HIV are only speculative as the authors did not infect the mice with HIV. There should be a consideration of the weaknesses of rodent models for HIV pathogenesis and how this impacts the presentation of other pathologies associated with HIV infection. Why did the authors only focus on immunoanalyses of innate immune cells? Especially given that they proceed to use Rag-1 deficient mice which have defective adaptive immune systems? What would be the importance of the significantly lower eosinophils in the Rag 1-/- mice that had worse disease? Also, it is noted that the cytokines are produced a week after infections despite a large fungal burden. How is this mediated and why is the response so delayed? Is there another player of the adaptive immune system that may be triggering these responses in the lung? How closely does the widespread infection in mice (in multiple tissues) compare to the human case? Was this a hyperbolic response? If so, how can one interpret results from these models in a human context? Was there a reason humanized mice (with a reconstitution of the human immune systems) were not used in these studies? What about the immune response in other tissues? Of interest is the persistent infection in the spleen, an immunoregulatory site... why was this evident? Some speculation on this would be of interest. A lot of comparisons are made with H. capsulatum infections in the discussion. These comparisons confuse the reader and sometimes I am not sure if the paper was to establish a murine model of E. africanus infections or to compare and contrast its pathology to known H. capsulatum infections. Why would we care about H. capsulatum infections anyway in the context of advanced HIV? Most patients with advanced HIV are also on ARVs. What would be the impact of ARV treatment on E. africanus infections?

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Reviewer #1: No

Reviewer #2: No

Figure Files:

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References

Please review your reference list to ensure that it is complete and correct. If you have cited papers that have been retracted, please include the rationale for doing so in the manuscript text, or remove these references and replace them with relevant current references. Any changes to the reference list should be mentioned in the rebuttal letter that accompanies your revised manuscript. If you need to cite a retracted article, indicate the article's retracted status in the References list and also include a citation and full reference for the retraction notice.

Dear Mrs. Hoving,

We are pleased to inform you that your manuscript 'The pathogenesis of experimental Emergomycosis in mice.' 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,

Marcio L Rodrigues

Section Editor

PLOS Neglected Tropical Diseases

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