Differential immunomodulation of porcine bone marrow derived dendritic cells by E. coli Nissle 1917 and β-glucans

Mirelle Geervliet; Laura C. P. Lute; Christine A. Jansen; Victor P. M. G. Rutten; Huub F. J. Savelkoul; Edwin Tijhaar

doi:10.1371/journal.pone.0233773

Peer Review History

Original SubmissionOctober 10, 2019
18 Feb 2020 Decision Letter - Miquel Vall-llosera Camps, Editor PONE-D-19-28012 Differential immunomodulation of porcine bone marrow derived dendritic cells by E. coli Nissle 1917 and β-glucans PLOS ONE Dear Mrs Geervliet, 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. I would like to sincerely apologise for the delay you have incurred with your submission. It has been exceptionally difficult to secure reviewers to evaluate your study. We have now received three completed reviews; their comments are available below. Reviewer#1 and #3 have raised some concerns about the study that need to be addressed in a revision. Both reviewers have noted that a clearer rationale for your study should be provided. Please revise the manuscript to address all the reviewer's comments in a point-by-point response in order to ensure it is meeting the journal's publication criteria. Please note that the revised manuscript will need to undergo further review, we thus cannot at this point anticipate the outcome of the evaluation process. We would appreciate receiving your revised manuscript by Apr 02 2020 11:59PM. When you are ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file. If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. To enhance the reproducibility of your results, we recommend that if applicable you deposit your 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/plosone/s/submission-guidelines#loc-laboratory-protocols Please include the following items when submitting your revised manuscript: A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). This letter should be uploaded as separate file and labeled 'Response to Reviewers'. A marked-up copy of your manuscript that highlights changes made to the original version. This file should be uploaded as separate file and labeled 'Revised Manuscript with Track Changes'. An unmarked version of your revised paper without tracked changes. This file should be uploaded as separate file and labeled '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. We look forward to receiving your revised manuscript. Kind regards, Miquel Vall-llosera Camps Associate Editor PLOS ONE Journal Requirements: When submitting your revision, we need you to address these additional requirements. 1. Please ensure that your manuscript meets PLOS ONE's style requirements, including those for file naming. The PLOS ONE style templates can be found at http://www.journals.plos.org/plosone/s/file?id=wjVg/PLOSOne_formatting_sample_main_body.pdf and http://www.journals.plos.org/plosone/s/file?id=ba62/PLOSOne_formatting_sample_title_authors_affiliations.pdf 2. To comply with PLOS ONE submissions requirements, please provide methods of sacrifice in the Methods section of your manuscript. 3. We note that you have included the phrase “data not shown” in your manuscript. Unfortunately, this does not meet our data sharing requirements. PLOS does not permit references to inaccessible data. We require that authors provide all relevant data within the paper, Supporting Information files, or in an acceptable, public repository. Please add a citation to support this phrase or upload the data that corresponds with these findings to a stable repository (such as Figshare or Dryad) and provide and URLs, DOIs, or accession numbers that may be used to access these data. Or, if the data are not a core part of the research being presented in your study, we ask that you remove the phrase that refers to these data. 4. We note that you have stated that you will provide repository information for your data at acceptance. Should your manuscript be accepted for publication, we will hold it until you provide the relevant accession numbers or DOIs necessary to access your data. If you wish to make changes to your Data Availability statement, please describe these changes in your cover letter and we will update your Data Availability statement to reflect the information you provide. [Note: HTML markup is below. Please do not edit.] Reviewers' comments: Reviewer's Responses to Questions 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 Reviewer #2: Yes Reviewer #3: Yes ******** 2. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: I Don't Know Reviewer #2: Yes Reviewer #3: Yes ****** 3. 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 Reviewer #2: Yes Reviewer #3: Yes ****** 4. 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 Reviewer #2: Yes Reviewer #3: Yes ****** 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: Authors describe an experiment designed study the possible immunomodulating effects of EcN and betaglucans on dendritic cells. It is in principle a study worth publishing with interesting results. There are however a few points to be resolved before publication. In the introduction it remains unclear what exactly is meant by immunomodulation and what the authors expect from it in terms of animal health. As is, it is very vague. For instance, do the authors aim for higher titers, stronger inflammatory responses etc., and would that lead to better health? Also, the results obtained should be discussed in the light of the above. It is currently not clear from the discussion what the results actually mean for the animals. The other important point is that the information about betaglucans is too summary. There are many different types and sources of betaglucans, and different properties and immunomodulating effects have been ascribed to them e.g. soluble vs particulate etc. This should be at least briefly be mentioned in the introduction. Furthermore, it is not described what type of betaglucan was used (soluble? DP?) and what the other 40% consists of. Reviewer #2: I this study, authors assess the immunomodulatory potential of E. coli Nissle 1917 and β-glucans in vitro using porcine bone marrow-derived dendritic cell. The finding of this study are interesting, demonstrated that both β-glucans and E. coli Nissle 1917 were able to enhance dendritic cell maturation, but in a differential manner. In my opinion the manuscript is well structured and well written with minor topographical errors. I recommend to accept this paper. Reviewer #3: This is a methodologically sound study describing the effects of foodstuffs, based on a strain of E. coli and Beta-glucans, for their effects on dendritic cells (DC), as modeled by DC derived in vitro from bone marrow cells. I presume that it the commercial use of these particular materials that has led to their selection for this study but this isn't very explicit. There is a sound rationale for focusing on DC in this sort of analysis but a major limitation of this sort of study is that it is unclear what sort of DC is relevant, what defines a 'good' response in DC and what are the circumstances in which these responses can be induced in the intestine in vivo. Perhaps these issues could be touched upon the manuscript. I would make the following addition minor comments and suggestions: - What was the control for the CD152-Ig staining reagent? - How Were MFI values calculated ? Was the MFI for the control staining subtracted? - It would be good to show example FACS plots for the relevant markers in stimulated cells in the main body of the paper (so the reader can get an idea of how the MFI values shown in graphs were derived. - Figure legends should consistently state the number of replicates/independent experiments. It is recognised that the nature of work on pigs mean that replication will be limited - but it needs to be explicit. - There are some minor typos etc:'rpm' instead of 'g' (L153); lack of superscript (L119); Figure S4 is not referred to in the text; the figure panels in S2 are not explained in the legend; what is FCM (L307)? - Clarify what 'remains to be detected' (L399) means? There is no data? Ambiguous data? Conflicting data? - What does 'appeared' (L266) mean? - The values of the PCA analysis is unclear. It isn't really explained or described in the Results section ( there is some attempt to address this belatedly in Discussion) and it feels a bit of an afterthought. ****** 6. 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 Reviewer #3: No [NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files to be viewed.] 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. Registration is free. 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. Please note that Supporting Information files do not need this step. https://doi.org/10.1371/journal.pone.0233773.r001
Revision 1
2 Apr 2020 Author Response Dear editor, We would like to thank you and the reviewers for the valuable comments on our manuscript entitled “Differential immunomodulation of porcine bone marrow derived dendritic cells by E. coli Nissle 1917 and β-glucans” by Mirelle Geervliet1, Laura C.P. Lute1, Christine A. Jansen2, Victor P.M.G. Rutten2,3, Huub F.J. Savelkoul1, Edwin Tijhaar1. We have carefully read the comments that were presented and we resubmitted our paper including revisions. Please note that the line numbers (in yellow) correspond to the line numbers in the revised document. Below we give a detailed point-by-point reply regarding the comments and we adapted the manuscript accordingly. ACADEMIC EDITOR SPECIFIC COMMENTS: 1. Please ensure that your manuscript meets PLOS ONE's style requirements, including those for file naming. The PLOS ONE style templates can be found at: - http://www.journals.plos.org/plosone/s/file?id=wjVg/PLOSOne_formatting_sample_main_body.pdf - http://www.journals.plos.org/plosone/s/file?id=ba62/PLOSOne_formatting_sample_title_authors_affiliations.pdf Authors response: we have adopted the requirements according to your suggestions in the revised manuscript. The file names are adjusted according to the PLOSONE requirements. 2. To comply with PLOS ONE submissions requirements, please provide methods of sacrifice in the Methods section of your manuscript. Authors response: The methods of sacrifice is now included in the Methods section (Ethics statement) of the manuscript (Lines 129 – 130). 3. We note that you have included the phrase “data not shown” in your manuscript. Unfortunately, this does not meet our data sharing requirements. PLOS does not permit references to inaccessible data. We require that authors provide all relevant data within the paper, Supporting Information files, or in an acceptable, public repository. Please add a citation to support this phrase or upload the data that corresponds with these findings to a stable repository (such as Figshare or Dryad) and provide and URLs, DOIs, or accession numbers that may be used to access these data. Or, if the data are not a core part of the research being presented in your study, we ask that you remove the phrase that refers to these data. Authors response: We removed the phrase ‘data not shown’ from the manuscript and referred to the public repository accordingly. 4. We note that you have stated that you will provide repository information for your data at acceptance. Should your manuscript be accepted for publication, we will hold it until you provide the relevant accession numbers or DOIs necessary to access your data. If you wish to make changes to your Data Availability statement, please describe these changes in your cover letter and we will update your Data Availability statement to reflect the information you provide. Authors response: Thank you for this information. We have now obtained the requested repository information and DOI number. We removed the phrase ‘data not shown’ from the manuscript and referred to the public repository in lines 223, 254, 257 and 288. Therefore no changes regarding our data availability statement. REVIEWER COMMENTS: Reviewer #1: Authors describe an experiment designed study the possible immunomodulating effects of EcN and betaglucans on dendritic cells. It is in principle a study worth publishing with interesting results. There are however a few points to be resolved before publication. • In the introduction it remains unclear what exactly is meant by immunomodulation and what the authors expect from it in terms of animal health. As is, it is very vague. For instance, do the authors aim for higher titers, stronger inflammatory responses etc., and would that lead to better health? Authors response: We reformulated the text in the introduction (Lines 50 - 59) and discussion (Lines 404 - 409 and Lines 481 - 483) to make clear what we mean with ‘immunomodulation’; a more primed/mature state of the immune system that may contribute to a better resistance against (bacterial and viral) infections. In addition, we explain how maturation of DCs could benefit this (Lines 59 - 61). What it could mean in terms of animal health is addressed at the end of the abstract (Lines 38 - 39) and at the end of the discussion (Lines 481 - 483). • Also, the results obtained should be discussed in the light of the above. It is currently not clear from the discussion what the results actually mean for the animals. Authors response: As mentioned above, at the end of the abstract (Lines 38 - 40) and at the end of the discussion (Lines 481 - 483) it is better explained what the results of this study could mean for animal health. • The other important point is that the information about betaglucans is too summary. There are many different types and sources of betaglucans, and different properties and immunomodulating effects have been ascribed to them e.g. soluble vs particulate etc. This should be at least briefly be mentioned in the introduction. Authors response: This is indeed important to address in this article. We added additional lines about the different types and properties of betaglucans (with references) in the introduction (Lines 77 - 87). • Furthermore, it is not described what type of betaglucan was used (soluble? DP?) and what the other 40% consists of. Authors response: The type of betaglucans used (particulate/structure) and the composition of MacroGard® (percentage), including the non-betaglucan part, is now described in Lines 77 - 78 and Lines 160 - 163. Reviewer #3: This is a methodologically sound study describing the effects of foodstuffs, based on a strain of E. coli and Beta-glucans, for their effects on dendritic cells (DC), as modeled by DC derived in vitro from bone marrow cells. • I presume that it the commercial use of these particular materials that has led to their selection for this study but this isn't very explicit. Authors response: The promising immunomodulatory properties of MacroGard (yeast-derived and particulate beta-glucans) and its commercial use in pig feed has led to the selection of this type of beta-glucans. This is now clarified in the introduction with references (lines 81 – 87). EcN is known as a versatile probiotic interacting with cells of the immune system including antigen presenting cells such as DCs. Despite its (commercial) use over 100 years in humans there is only limited information of its immunomodulatory effect in pigs (Lines 87 – 93), prompting us to examine its effect on porcine DCs. • There is a sound rationale for focusing on DC in this sort of analysis but a major limitation of this sort of study is that it is unclear what sort of DC is relevant, what defines a 'good' response in DC and what are the circumstances in which these responses can be induced in the intestine in vivo. Perhaps these issues could be touched upon the manuscript. Authors response: These are indeed valid points to add to the discussion. There are different types of DCs which possess different characteristic and functions based on their location and maturation status. Intestinal DCs are considered to be the target for β-glucan and probiotic activity in vivo, but are not easily accessible for in vitro studies. We have now added a more extensive explanation on this point (Lines 411 – 414). It is indeed difficult to define what is a ‘good’ DC response. Higher expression levels of the co-stimulatory molecules CD80/86 on antigen presenting cells implies that these cells are more likely to reach the required threshold for T-cell activation. In addition, the production of cytokines by the antigen presenting cells can also contribute to the development of a more effective immune response against the presented antigen (Lines 404 - 409). The circumstances in which these responses can be induced in the intestine in vivo is briefly mentioned in Lines 59 - 61 and Lines 411-412. We realize what limitations in vitro studies can have for the situation in vivo. We consider this in vitro model as a valuable method to pre-screen putative feed additives, and to select the more promising ones that will then have to be validated in vivo. This is now indicated in Lines 481 – 483. I would make the following addition minor comments and suggestions: • What was the control for the CD152-Ig staining reagent? Authors response: Thank you for this question. Different controls have been used in this study. Firstly, single color stains for setting the compensation have been described in the Materials and Methods section of the manuscript. Secondly, we used unstained cells to detect possible autofluorescence. We noticed that this control accidently was missing in the manuscript, but is now included in Line 208. Because of the limited number of colors used (FITC, PE and APC) with very different emission wavelengths we did not include FMO controls. Retrospectively, the FMO control could have been included. Nonetheless, we are confident about the specific staining of the CTLA4Ig Fusion Protein as it is widely used and accepted in in vitro and in vivo studies in pigs and our CD80/86 staining/expression is in line with what has been described in other literature. We have added now added additional references (see references (1-3) below). In addition, Figure S2 shows that cells that express SLA Class-II also express CD80/86 as expected. Cells that do not express SLA Class-II do not express CD80/86 and thus no unspecific binding occurs. • How were MFI values calculated? Was the MFI for the control staining subtracted? Authors response: The MFI values were calculated for the CD172a+/high CD80/86+ and SLA Class-II+ population (Figure 2, panel 6). A new and more clear explanation is added in the Materials and Methods section (Lines 212 - 213). In addition, we adjusted the gating of the SLA Class-II/CD80/86 double positive population to make it more clear for the reader how the MFI was actually calculated (Figure 2, panel 6). The MFI of the control is not subtracted. The MFI of the control is indicated by the white bar in Figure 3, 4, 5 and 6 (and S4 and S5 Figure). • It would be good to show example FACS plots for the relevant markers in stimulated cells in the main body of the paper (so the reader can get an idea of how the MFI values shown in graphs were derived. Authors response: Thank you for this suggestion. We agreed that this addition will indeed add value to the manuscript. We added representative plots for the marker CD80/86 (Figure 5, panel A). In addition, we adjusted Figure 3 in order to show the effect of different concentrations of LPS. Both adjustments to the figure will help to better understand how the MFI values in the graphs were derived. • Figure legends should consistently state the number of replicates/independent experiments. It is recognised that the nature of work on pigs mean that replication will be limited - but it needs to be explicit. Authors response: We agree with this notion. The figure legends have been reviewed again and the number of replicates were now added when missing (Figure 3, 4, 7 and S4 Figure). • There are some minor typos etc:'rpm' instead of 'g' (L153); lack of superscript (L119); Figure S4 is not referred to in the text; the figure panels in S2 are not explained in the legend; what is FCM (L307)? Authors response: Thank you for noticing these typos. They are now corrected: - Line 179: ‘rpm’ adjusted to ‘g’ - Line 142: Superscript is now included - Figure S4: All Figures are referred to now. - Figure S2: The figure panels are now explained in the legend. - Line 192: With FCM I meant Flow Cytometry. As this abbreviation is not commonly used and to avoid confusion it has now been spelled out everywhere in the paper. • Clarify what 'remains to be detected' (L399) means? There is no data? Ambiguous data? Conflicting data? Authors response: To date, there is no data available. Line 399 (now Lines 432 - 434) has been re-written to clarify this. • What does 'appeared' (L266) mean? Authors response: The word ‘appeared’ is not used correctly in this context (too ambiguous). The word appeared has been removed. • The values of the PCA analysis is unclear. It isn't really explained or described in the Results section (there is some attempt to address this belatedly in Discussion) and it feels a bit of an afterthought. Authors response: Thank you for the comment. The reason to conduct the principal component analysis (PCA) was to analyse the grouping of these responses of frhBMDCs and cryoBMDCs to different stimulation conditions. We now explain the value of the PCA plot as these values are described and explained more extensively in the results section. The paragraph (Lines 368 - 386) has been re-written. In addition, the legend of Figure 7 has been re-written. To connect the PCA with the previous results, we have now clarified the added value of conducting such an analysis in Lines 368 – 372. AUTHOR COMMENTS: We made some minor changes which were detected during the review process. • In the article we used ‘Macrogard’ and ‘MacroGard’. The latter brand name is the correct one and we adjusted this throughout the article. • In the article we used ‘mean fluorescent intensity’ this should be ‘median fluorescent intensity’.We adjusted this throughout the article. • The figure legenda’s and the order of the figures are adjusted according to the PLOSONE’s style requirements. Looking forward to your decision. With kind regards and on behalf of the authors, Mirelle Geervliet Cell Biology and Immunology Group Wageningen University P.O. Box 338 6700 AH Wageningen The Netherlands E-mail: geervlietmirelle@gmail.com Telephone number: +3161657994 References: 1. Piriou-Guzylack L, Salmon H. Membrane markers of the immune cells in swine: an update. Veterinary research. 2008;39(6):1. 2. Summerfield A, McCullough KC. The porcine dendritic cell family. Developmental & Comparative Immunology. 2009;33(3):299-309. 3. Vreman S, Auray G, Savelkoul HF, Rebel A, Summerfield A, Stockhofe-Zurwieden N. Neonatal porcine blood derived dendritic cell subsets show activation after TLR2 or TLR9 stimulation. Developmental & Comparative Immunology. 2018;84:361-70. Attachments Attachment Submitted filename: Response to reviewers.docx* https://doi.org/10.1371/journal.pone.0233773.r002
13 May 2020 Decision Letter - Jagadeesh Bayry, Editor Differential immunomodulation of porcine bone marrow derived dendritic cells by E. coli Nissle 1917 and β-glucans PONE-D-19-28012R1 Dear Dr. Geervliet, 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. Shortly after the formal acceptance letter is sent, an invoice for payment will follow. To ensure an efficient production and billing process, please log into Editorial Manager at https://www.editorialmanager.com/pone/, click the "Update My Information" link at the top of the page, and update your user information. If you have any billing related questions, please contact our Author Billing department directly at authorbilling@plos.org. If your institution or institutions have a press office, please notify them about your upcoming paper to enable them to help maximize its impact. If they will be preparing press materials for this manuscript, you must inform our press team as soon as possible and no later than 48 hours after receiving the formal acceptance. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information, please contact onepress@plos.org. With kind regards, Jagadeesh Bayry, DVM, PhD, HDR 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 #2: All comments have been addressed Reviewer #3: All comments have been addressed ******** 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 #2: Yes Reviewer #3: Yes ****** 3. Has the statistical analysis been performed appropriately and rigorously? Reviewer #2: Yes Reviewer #3: 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 #2: Yes Reviewer #3: 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 #2: Yes Reviewer #3: 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 #2: Authors have made all suggested changes and improved the manuscript as required. I recommend the editorial board to accept this paper. Reviewer #3: I made several comments and suggestions on the original submission. I am satisfied that the authors have taken these comments on board and made changes to the manuscript accordingly. ****** 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. 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 #2: No Reviewer #3: No https://doi.org/10.1371/journal.pone.0233773.r003
Formally Accepted
10 Jun 2020 Acceptance Letter - Jagadeesh Bayry, Editor PONE-D-19-28012R1 Differential immunomodulation of porcine bone marrow derived dendritic cells by E. coli Nissle 1917 and β-glucans Dear Dr. Geervliet: I'm pleased to inform you that your manuscript has been deemed suitable for publication in PLOS ONE. Congratulations! Your manuscript is now with our production department. If your institution or institutions have a press office, please let them know about your upcoming paper now to help maximize its impact. If they'll be preparing press materials, please inform our press team within the next 48 hours. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information please contact onepress@plos.org. If we can help with anything else, please email us at plosone@plos.org. Thank you for submitting your work to PLOS ONE and supporting open access. Kind regards, PLOS ONE Editorial Office Staff on behalf of Dr. Jagadeesh Bayry Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0233773.r004

Open letter on the publication of peer review reports

PLOS recognizes the benefits of transparency in the peer review process. Therefore, we enable the publication of all of the content of peer review and author responses alongside final, published articles. Reviewers remain anonymous, unless they choose to reveal their names.

We encourage other journals to join us in this initiative. We hope that our action inspires the community, including researchers, research funders, and research institutions, to recognize the benefits of published peer review reports for all parts of the research system.

Learn more at ASAPbio .