Granular Insights on Innate and Intrinsic Immunity to Flaviviruses
Round 1
Reviewer 1 Report
Comments and Suggestions for AuthorsThis is a timely and insightful review addressing the important topic of emerging pathogens. However, the authors should include a detailed description of the methodology used for literature selection to enhance the transparency and reproducibility of the review. In addition, the section on Powassan virus is notably absent and should be incorporated to improve the comprehensiveness of the manuscript. Finally, it is recommended that each section conclude with a brief paragraph highlighting the clinical relevance of the discussed pathogen, which would enhance the review’s utility and appeal to a broader audience, including practicing clinicians.
I clicked that all references are appropriate
I also provided that authors should add methodology.
I also provided 2 very important suggestions
1) to add Powassan virus paragraph and
2) to add in each section clinical relevance
The authors conclusions are appropriate
Author Response
We appreciate the reviewer's comments. Responses to specific comments are below:
However, the authors should include a detailed description of the methodology used for literature selection to enhance the transparency and reproducibility of the review.
Response: We added the following statement on page 1. "To identify relevant articles for inclusion in this review, searchable databases (PubMed, Google Scholar) were used with search terms “flavivirus” and “stress granules”. The same databases were used to identify relevant background articles."
In addition, the section on Powassan virus is notably absent and should be incorporated to improve the comprehensiveness of the manuscript.
Response: We could not identify any relevant articles linking stress granules and Powassan virus. However, we now add a section on tick-borne flaviviruses, highlighting two relevant papers.
Finally, it is recommended that each section conclude with a brief paragraph highlighting the clinical relevance of the discussed pathogen, which would enhance the review’s utility and appeal to a broader audience, including practicing clinicians.
Response: We added text at the beginning of several sections on specific viruses concerning the clinical relevance of each pathogen. For ZIKV, the original text has a relevant description.
Reviewer 2 Report
Comments and Suggestions for AuthorsThis timely review provides a comprehensive synthesis of the complex relationship between stress granules (SGs) and flavivirus infections, with a valuable focus on Zika virus (ZIKV). The manuscript critically examines conflicting literature on SG roles in antiviral immunity (immune activation vs. viral RNA sequestration) and offers novel hypotheses. Strengths include the detailed analysis of ZIKV-SG interactions (Table 1) and integration of recent advances like viral aggregated RNA condensates (VARCs). However, the review would benefit from clearer articulation of its conceptual advancements over prior reviews (e.g., Valadao et al., 2016), deeper reconciliation of contradictory data, and tighter structural integration of key figures/hypotheses. With revisions, this will be a significant contribution to the field.
Introduction: Explicitly state how this review advances beyond Valadao et al. (2016). Highlight unique contributions (e.g., analysis of post-2017 controversies, VARC concept in flaviviruses, sfRNA hypothesis).
Figure 1: Define all abbreviations in the legend.
Table 1: Standardize "h.p.i." (hours post-infection) and "MOI" formatting.Table 1: Expand annotations to include:
Methodological variables (e.g., antibody clones for SG markers, criteria for "SG-positive" cells).
Cell-type specificities (e.g., IFN competence in A549 vs. Vero cells explaining differential PKR activation).
Abbreviations: List all in a footnote.
Section 11 (ZIKV): Add a synthesis paragraph reconciling disparities.
Author Response
We thank the reviewer for feedback and positive comments. The review has been revised accordingly. Specific response to the reviewer comments are below:
Introduction: Explicitly state how this review advances beyond Valadao et al. (2016). Highlight unique contributions (e.g., analysis of post-2017 controversies, VARC concept in flaviviruses, sfRNA hypothesis).
Response: Additional text has been added on page 1 to clarify how this review extends that by Valadao et al.
Figure 1: Define all abbreviations in the legend.
Response: All abbreviations are now defined.
Table 1: Standardize "h.p.i." (hours post-infection) and "MOI" formatting.Table 1: Expand annotations to include:
Methodological variables (e.g., antibody clones for SG markers, criteria for "SG-positive" cells).
Cell-type specificities (e.g., IFN competence in A549 vs. Vero cells explaining differential PKR activation).
Response: HPI and MOI notation is now standardized. A footnote has been added to the table to indicate other methodological variables that may impact experimental results.
Abbreviations: List all in a footnote.
Response: All abbreviations are now listed .
Section 11 (ZIKV): Add a synthesis paragraph reconciling disparities.
We feel the concluding paragraph attempts to reconcile disparities and add additional text indicating that reconciliation of available data is not feasible at this time. We add that future well-controlled and rigorous studies are needed to clarify the apparent disparities.
Round 2
Reviewer 1 Report
Comments and Suggestions for AuthorsI am unable to see in the revised text the changes that authors have made regarding clinical implications for each disease they described
Author Response
We apologize for the lack of clarity. For some reason, the revised text which was highlighted in red was not completely retained in the submitted revised manuscript. A summary of the relevant revisions follows below:
For WNV, we added the following new text:
WNV is an encephalitic mosquito-borne flavivirus transmitted by Culex mosquitos. Although humans are considered a dead-end host, WNV infection can cause serious neurological disease which may be fatal or lead to long-term sequelae68. Like most other flaviviruses, there are no available vaccines or therapeutics to combat WNV infections.
For HCV, we added the following text:
Chronic infection may lead to development of liver cirrhosis and hepatocellular carcinoma.
For tick-borne flaviviruses, we added the following text:
Tick-borne encephalitis virus (TBEV) and Powassan virus (POWV) are transmitted via ticks which have a much longer life span than mosquitos. As a result, tick-borne flaviviruses must persist for longer periods in their vector host than mosquito-borne viruses. This may be associated with reduced genomic variation in these viruses compared to mosquito-borne flaviviruses. Like WNV, both TBEV and POWV are encephalitic, and infection may lead to fatal outcome.
For ZIKV, we feel the existing text addresses clinical relevance:
ZIKV is a flavivirus of significant interest for multiple reasons, including its capacity for explosive spread in a naïve population as seen in the 2015-2016 American epidemic, its ability to transmit vertically and cause birth defects, and its ability to be sexually transmitted85. At the molecular level, ZIKV resembles other mosquito-borne flaviviruses in genome organization and virion structure. The biological basis for ZIKV’s unique pathogenesis features is not well understood.
We agree with the reviewer that addition of text addressing clinical relevance of these pathogens improves the manuscript. However, we feel that extensive description of clinical relevance is beyond the scope of this review which strongly focuses on cellular and molecular biology of flavivirus infections.
We thank the reviewer for their time and effort to review our manuscript.
Best,
Shelton
Reviewer 2 Report
Comments and Suggestions for AuthorsI appreciate the authors' thorough responses; my concerns are now fully resolved. This review critically examines the complex and often conflicting roles of stress granules in restricting Flaviviridae viruses like Zika, deepening our understanding of their dual antiviral mechanisms (immune signaling hub and intrinsic RNA sequestration) and providing valuable clues for developing novel host-directed antiviral strategies.
Author Response
We thank the reviewer for their time and effort to review our manuscript.
Best,
Shelton
