Peer Review History
Original SubmissionNovember 26, 2021 |
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PONE-D-21-37491Inhibition of experimental autoimmune uveitis by intravitreal AAV-Equine-IL10 gene therapyPLOS ONE Dear Dr. Crabtree, 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. There are several critical issues raised by the reviews that include missing information in the methods and in the experimental approach. Please submit your revised manuscript by Apr 24 2022 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org. When you're 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. Please include the following items when submitting your revised manuscript:
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[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: Partly Reviewer #2: Partly ********** 2. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: I Don't Know Reviewer #2: 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 ********** 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 ********** 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: The authors address the significant unmet need for an effective, local, non-corticosteroid treatment option for non-infection uveitis. The goal of the study is to use a rat model of uveitis to study feasibility of their gene therapy approach prior to use in equine recurrent uveitis (ERU). The authors test the ability of two doses of an AAV8 vector expressing IL-10 to prevent EAU in Lewis rats. They conclude that intravitreal injection of both low and high dose vectors significantly decreased ocular inflammation and retinal damage. The manuscript is well written. The intro and discussion are thorough, and figures are appropriate. There are a few experimental design concerns that limit the significance of the conclusions. Specifically, that the treatment effect is local, and that the treatment approach is sufficient to treat uveitis in the presence of an established anti-retinal T cell population. Despite these concerns, this is an interesting study. Experimental design: 1. Immunosuppression administered prior to or during the immunization phase of EAU can prevent development of the anti-ocular T cell population. • In this study, it would be important to show that the local treatment approach (administered prior to uveitis induction) did not prevent the development of the expected autoreactive T cells. Ultimately for this gene therapy approach to be effective it must function in the context of the anti-ocular effector T cell population. Currently that data could also be interpreted to mean that the treatment prevented the generation of the anti-ocular T cell population at the time of immunization. o Optional experiments would be to treat rats with IVT AAV8-IL-10 and then use adoptive transfer to generate EAU. Alternatively, T cell ELISPOT from immunized rats could be used to demonstrate the autoreactive T cells were present. 2. With the assumption that an autoreactive T cell population was generated as expected (see above concern), it is not clear that the beneficial effect of the gene therapy is local to the eye. The low dose treatment showed the same benefit as high dose treatment, but without similar evidence of local gene expression. An alternative hypothesis is that increased systemic IL-10 expression (in eye draining local lymph nodes or the spleen for example) could be responsible for the beneficial treatment effect. This could be addressed in the discussion and further mechanistic studies. Additional experiments to address this concern would be beyond the scope of this proof-of-concept study. Statistical question: • Was clinical score data (Figure 1) analyzed per eye or per animal? Please clarify. If per eye, there needs to be an adjustment to account for non-independence of eye level data, or score could be assigned per animal by averaging both eyes. Comment: Cell mediated killing of transduced cells is a concern for the safety and efficacy of gene therapy approaches. I was stuck by the appearance of the ciliary body in the high dose treatment group. There looks to have been considerable loss of CB tissue and thinning of the iris. Was this seen in multiple eyes, or is just this section abnormal? Reviewer #2: This manuscript reports intravitreal administration of an IL-10-AAV8 vector 7 days prior to inducing EAU in Lewis Rats, and the authors were able to detect IL-10 transcripts in ocular tissues and found a reduction in EAU and in the inflammatory cell infiltration in the IL-10-AAV8-treated EAU rats relative to BSS-injected EAU controls. They conclude this to be an effective therapy which should be considered for treating equine recurrent uveitis. Comments: 1. How many experimental replicates were performed in this study? It would appear from the text that only one experiment was carried out with 14 rats divided into three treatment groups. Due to the variability in EAU in rodent models, it is expected that any treatment effect be reproduced at least 3 times. Can the authors please clarify in the Results how many repeat experiments were carried out. 2. The use of the rat model in interesting as it is generally an acute form of ocular inflammation which eventually burns out over the course of a few weeks. Did the authors consider using a more recurrent form of EAU in mice? This might potentially be a more representative model with which to test this novel therapy. 3. Given that rat EAU is a very acute, and clinically severe form of uveitis, it is nevertheless impressive that a suppression of EAU was observed using both doses of vector. Did the authors consider administering the treatment 7 days AFTER inducing EAU? This is a commonly used approach to determine whether a treatment can down regulate an already established disease, and should be discussed in respect of the current study. 4. The histology images in Fig 3 are too faint in my version to easily detect the infiltrating cells, and should be taken at a higher resolution. The retinal layers appear relatively unaffected, but have become detached from the choroid. Is this an artefact of the fixation proceedure, or an effect of the EAU? 5. Whilst I agree that it is good to detect IL-10 in the ciliary body and iris, the blood-retinal barrier at the level of the retinal endothelial cells is also a key part of the barrier. It would be helpful if semi-quantitative data for the IL-10 PCR in each tissue was shown. Data in Fig 4 is difficult to understand, as there is no reference to IL-10, only vector cDNA. Whilst I appreciate that the safety issues were of huge concern, it is also important to know whether, at the larger concentration, IL-10 could in fact be detected within the retinal tissues. ********** 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? 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Revision 1 |
Inhibition of experimental autoimmune uveitis by intravitreal AAV-Equine-IL10 gene therapy PONE-D-21-37491R1 Dear Dr. Gilger, We’re pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements. Within one week, you’ll receive an e-mail detailing the required amendments. When these have been addressed, you’ll receive a formal acceptance letter and your manuscript will be scheduled for publication. An invoice for payment will follow shortly after the formal acceptance. To ensure an efficient process, please log into Editorial Manager at http://www.editorialmanager.com/pone/, click the 'Update My Information' link at the top of the page, and double check that your user information is up-to-date. 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 help maximize its impact. If they’ll be preparing press materials, please inform our press team as soon as possible -- 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. Kind regards, Andrew W Taylor, Ph.D. 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 ********** 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 ********** 3. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: 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 #1: 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 #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 have been very responsive to reviews. My concerns have been addressed. Statistical concerns have been clarified and new histology images addressed my concerns about a toxic effect of treatment. ********** 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 #1: Yes: Kathryn Pepple, MD, PhD ********** |
Formally Accepted |
PONE-D-21-37491R1 Inhibition of experimental autoimmune uveitis by intravitreal AAV-Equine-IL10 gene therapy Dear Dr. Gilger: 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. Andrew W Taylor Academic Editor PLOS ONE |
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