https://orcid.org/0000-0001-7424-5383
Peer Review History
| Original SubmissionOctober 4, 2020 |
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Dear Prof. López-Botet, Thank you very much for submitting your manuscript "NK cells eliminate Epstein-Barr virus bound to B cells through a specific antibody-mediated uptake" for consideration at PLOS Pathogens. As with all papers reviewed by the journal, your manuscript was reviewed by members of the editorial board and by several independent reviewers. In light of the reviews (below this email), we would like to invite the resubmission of a significantly-revised version that takes into account the reviewers' comments. We cannot make any decision about publication until we have seen the revised manuscript and your response to the reviewers' comments. Your revised manuscript is also likely to be sent to reviewers for further evaluation. When you are ready to resubmit, please upload the following: [1] A letter containing a detailed list of your responses to the review comments and a description of the changes you have made in the manuscript. Please note while forming your response, if your article is accepted, you may have the opportunity to make the peer review history publicly available. The record will include editor decision letters (with reviews) and your responses to reviewer comments. If eligible, we will contact you to opt in or out. [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). Important additional instructions are given below your reviewer comments. Please prepare and submit your revised manuscript within 60 days. If you anticipate any delay, please let us know the expected resubmission date by replying to this email. Please note that revised manuscripts received after the 60-day due date may require evaluation and peer review similar to newly submitted manuscripts. Thank you again for your submission. 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, Erik K Flemington Section Editor PLOS Pathogens Erik Flemington Section Editor PLOS Pathogens Kasturi Haldar Editor-in-Chief PLOS Pathogens orcid.org/0000-0001-5065-158X Michael Malim Editor-in-Chief PLOS Pathogens *********************** Reviewer's Responses to Questions Part I - Summary Please use this section to discuss strengths/weaknesses of study, novelty/significance, general execution and scholarship. Reviewer #1: The authors demonstrate that natural killer (NK) cells prevent B cell infection by the Epstein Barr virus (EBV) in the presence of anti-viral antibody containing serum that does not inhibit infection on its own. In part this seems due to uptake of the viral particles by NK cells into early and then late endosomes. In the process some B cell markers are also transferred by trogocytosis, including CD21 the receptor for EBV attachment to B cells. The authors speculate that NK cells might limit EBV infection of B cells by non-lytic means in submucosal secondary lymphoid tissues like tonsils. This is an interesting proposition, but it remains unclear if this is a feature of all Fc receptor carrying cells and if it is somehow involved in NK cell infection by EBV. Reviewer #2: This paper demonstrates antibody dependent uptake of EBV viral particles in to NK cells after direct interaction with infected B cells. A number of different methods, including detection of viral gp350, GFP tagged particles, viral DNA and PKH26 labelled virus particles to detect transfer of EBV components from B cells and uptake by NK cell. The authors also demonstrate a reciprocal loss of EBV components from donor B cells. The experiments are mostly well controlled, the data is well analysed and presented and the conclusions are largely supported by the data. This study is novel as it potentially demonstrate a novel mechanism whereby NK cells could limit EBV infection in host B cells. ********** Part II – Major Issues: Key Experiments Required for Acceptance Please use this section to detail the key new experiments or modifications of existing experiments that should be absolutely required to validate study conclusions. Generally, there should be no more than 3 such required experiments or major modifications for a "Major Revision" recommendation. If more than 3 experiments are necessary to validate the study conclusions, then you are encouraged to recommend "Reject". Reviewer #1: 1. The authors suggest that the observed mechanism might be a new effector function of NK cells, but it remains unclear, if similar effects could be observed for all Fc receptor expressing cells. Are NK cells better than monocytes in removing EBV particles from B cell surfaces? NK cells should be compared in this respect with some classical phagocyte population. 2. Chronic active EBV infection (CAEBV) often spreads to other lymphocytes than B cells including NK cells. Could the observed mechanism facilitate NK cell infection by EBV? Any sign of latent EBV gene expression in NK cells after the antibody mediated uptake? As an alternative to latent EBV gene expression, GFP expression from recombinant EBV could be monitored in NK cells that have taken up EBV particles. 3. The authors observed CD19, CD20 and CD21 transfer to NK cells by trogocytosis. Is this of any functional consequence? Can CD21 expression and MHC class II up-regulation on activated NK cells be used for EBV infection? Is such infection blocked by antibodies against these respective surface markers? Reviewer #2: 1. For flow cytometric analysis (Figures 2, 5, 6, 7 ) the authors should demonstrate with some rigour (Other than comparison with the Rituximab control) how they can exclude that a small proportion of transient B-NK cell conjugates are not being detected within the NK cell gate thereby detecting B cell markers CD19, CD20 and CD21 and also virus gp350 labelled B cells within this gate. 2. How are small populations of dead cells being excluded from the co-culture systems? 3. Fig 7. Some clarification is needed for the data in figure 7A and 7B. In 7A, the right hand plot suggests a gradual attrition of virus from B cells with or without immune sera in the absence of NK cells. Does the co-culture system not simply reflect an acceleration of the process? The attrition of S+ in the absence of NK cells implies that this process is, at least in part, passive. Significance values should also be added to these plots to compare values across the time frame and between S- and S+ data points. 4. Figure 7D and E and supplementary video microscopy data. Although the this data does support uptake of viral components, what is the cumulative frequency of PKH26+ NK cells detected in these systems at any given point across the entire time frame and how does this compare to the steady state levels detected by flow cytometry (See point 1, above). 5. The authors demonstrate the co- localisation of EBV associated components with NK cell endosomal and lysosomal markers. Can they exclude the persistence of viable viral particles? This point should also be discussed in the context of evidence for or against a role for antibody facilitating active infection of NK cells. ********** Part III – Minor Issues: Editorial and Data Presentation Modifications Please use this section for editorial suggestions as well as relatively minor modifications of existing data that would enhance clarity. Reviewer #1: 1. The authors suggest that uncontrolled EBV infection in CD16A deficient patients points towards the importance of the described mechanism for EBV specific immune control in vivo. However, they should acknowledge that it was proposed that CD16A is required for CD2 function on NK cells outside of ADCC. Reviewer #2: 1. Fig 6. Could this be re-ordered to separate the subplots according to experimental conditions and to label the culture conditions more clearly. ********** 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: Yes: Martin R Goodier Figure Files: While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com. PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email us at figures@plos.org. Data Requirements: Please note that, as a condition of publication, PLOS' data policy requires that you make available all data used to draw the conclusions outlined in your manuscript. Data must be deposited in an appropriate repository, included within the body of the manuscript, or uploaded as supporting information. This includes all numerical values that were used to generate graphs, histograms etc.. For an example see here on PLOS Biology: http://www.plosbiology.org/article/info%3Adoi%2F10.1371%2Fjournal.pbio.1001908#s5. Reproducibility: To enhance the reproducibility of your results, PLOS recommends that you deposit laboratory protocols in protocols.io, where a protocol can be assigned its own identifier (DOI) such that it can be cited independently in the future. For instructions see http://journals.plos.org/plospathogens/s/submission-guidelines#loc-materials-and-methods |
| Revision 1 |
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Dear Dr López-Botet We are pleased to inform you that your manuscript 'NK cells eliminate Epstein-Barr virus bound to B cells through a specific antibody-mediated uptake' has been provisionally accepted for publication in PLOS Pathogens. 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 Pathogens. Best regards, Claire Shannon-Lowe Guest Editor PLOS Pathogens Erik Flemington Section Editor PLOS Pathogens Kasturi Haldar Editor-in-Chief PLOS Pathogens orcid.org/0000-0001-5065-158X Michael Malim Editor-in-Chief PLOS Pathogens *********************************************************** Reviewer Comments (if any, and for reference): |
| Formally Accepted |
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Dear Prof. López-Botet, We are delighted to inform you that your manuscript, "NK cells eliminate Epstein-Barr virus bound to B cells through a specific antibody-mediated uptake," has been formally accepted for publication in PLOS Pathogens. We have now passed your article onto the PLOS Production Department who will complete the rest of the pre-publication process. All authors will receive a confirmation email upon publication. The corresponding author will soon be receiving a typeset proof for review, to ensure errors have not been introduced during production. Please review the PDF proof of your manuscript carefully, as this is the last chance to correct any scientific or type-setting errors. Please note that major changes, or those which affect the scientific understanding of the work, will likely cause delays to the publication date of your manuscript. Note: Proofs for Front Matter articles (Pearls, Reviews, Opinions, etc...) are generated on a different schedule and may not be made available as quickly. Soon after your final files are uploaded, the early version of your manuscript, if you opted to have an early version of your article, will be published online. The date of the early version will be your article's publication date. The final article will be published to the same URL, and all versions of the paper will be accessible to readers. Thank you again for supporting open-access publishing; we are looking forward to publishing your work in PLOS Pathogens. Best regards, Kasturi Haldar Editor-in-Chief PLOS Pathogens orcid.org/0000-0001-5065-158X Michael Malim Editor-in-Chief PLOS Pathogens |
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