PONE-D-19-29134

High-mobility group box (TOX) antibody a useful tool for the identification of B and T cell subpopulation

PLOS ONE

Dear Dr Roncador,

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.

Please see reviewer comments below. Additionally, particular emphasis must be paid to the quality of the Western blot data and it's discussion within the text. For example, Fig.2A shows only a single band for TOX, why? Is it that HEK293 cells do not post-translationally alter the protein as suggested? I also note that reference 3 cited in the manuscript never ran TOX from their 293 cells on a Western, so this does not help clarify the observed anomaly. 

Please also note the journal's relatively new policy on cropped images, whereby the original, unmodified, images for all westerns and gels must now be submitted in the supplementary data.

Finally, the conclusions drawn from Fig.3 are invalid. You can not compare TOX expression between cell types whose loading controls are so vastly different. For example, to suggest RAJI express TOX while AKATA does not is simply unjustified. Indeed, loading controls are meant to be the same to show that direct comparisons can be made. I would ask the authors to repeat these Westerns to achieve the same loading levels before drawing any conclusions.

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PLOS ONE

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Additional Editor Comments (if provided):

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Reviewers' comments:

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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: 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

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Reviewer #1: In the present study, the authors investigated TOX expression in normal and neoplastic

lymphoid tissues using a novel rat monoclonal antibody that recognizes its target

molecule in paraffin-embedded tissue sections. They found that the majority of precursor B/T lymphoblastic, follicular and diffuse large B-cell lymphomas, nodular lymphocyte-predominant Hodgkin lymphomas and angioimmunoblastic T-cell lymphomas strongly expressed the TOX protein. Burkitt and mantle cell lymphomas showed TOX expression in a small percentage of cases. TOX was not found in the majority of chronic lymphocytic leukemia, myelomas, marginal zone lymphomas and classical Hodgkin lymphomas. A co-expression of TOX in PD-1+ CD8 T cells suggested a possible role of TOX in T-cell exhaustion. The authors concluded that TOX differential expression may be of diagnostic relevance in the differential diagnosis of follicular lymphoma, the identification of the phenotype of diffuse large B-cell lymphoma and the recognition of peripheral T-cell lymphoma with a follicular helper T phenotype.

Please note the following:

- The authors reported expression of TOX in BL cell line RAMOS and RAJI, but this latter data was not clearly represented in figure 3

- The study demonstrated expression pf TOX in germinal center B cells, however Burkitt lymphoma resulted TOX negative in most of the cases, although it’ s a GC derived tumor. This data should be highlighted in the discussion section.

- 65% of FL turned out to be TOX+, the authors should report the percentage of positive cases in each grade (1/2, 3A, 3B)

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Reviewer #1: No

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High-mobility group box (TOX) antibody a useful tool for the identification of B and T cell subpopulation

PONE-D-19-29134R1

Dear Dr. Roncador,

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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With kind regards,

Simon J Clark, D.Phil.

Academic 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

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

Reviewer #1: N/A

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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

**********

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 responded adequately to the reviewer's suggestions, modifying figure 3 and expanding the results and discussion section

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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.

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Reviewer #1: No