Binding of respiratory syncytial virus particles to platelets does not result in their degranulation in vitro
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Respiratory syncytial virus (RSV) is a major cause of severe respiratory infection in infants and the elderly. The mechanisms behind severe RSV disease are incompletely understood, but a dysregulated immune response probably plays an important role. Platelets are increasingly being recognized as immune cells and are involved in the pathology of several viruses. The release of chemokines from platelets upon activation may attract, for example, neutrophils to the site of infection, which is a hallmark of RSV pathology. In addition, since RSV infections are sometimes associated with cardiovascular events and platelets express several known RSV receptors, we investigated the effect of RSV exposure on platelet degranulation. Washed human platelets were incubated with sucrose-purified RSV particles. P-selectin and CD63 surface expression and CCL5 secretion were measured to assess platelet degranulation. We found that platelets bind and internalize RSV particles, but this does not result in degranulation. Our results suggest that platelets do not play a direct role in RSV pathology by releasing chemokines to attract inflammatory cells.
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The manuscript is improved following the author's revision and is now acceptable for publication in Access Microbiology
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Comments to Author
Comments on Lakerveld et al This manuscript focusses on respiratory syncytial virus (RSV), A clinical challenge public health risk without significant treatment. The authors investigate a very well justified aspect of the immune response to RSV and show, convincingly, that platelets are unlikely to play a role in RSV infection. Minor comments for the authors to address; Line 67; Please include a brief summary of the possibility that the gender of the platelet donors may affect the outcome of the study, however unlikely this may be and provide a suitable reference to lead the reader to the data on gender effects of platelets.
Please rate the manuscript for methodological rigour
Very good
Please rate the quality of the presentation and structure of the manuscript
V…
Comments to Author
Comments on Lakerveld et al This manuscript focusses on respiratory syncytial virus (RSV), A clinical challenge public health risk without significant treatment. The authors investigate a very well justified aspect of the immune response to RSV and show, convincingly, that platelets are unlikely to play a role in RSV infection. Minor comments for the authors to address; Line 67; Please include a brief summary of the possibility that the gender of the platelet donors may affect the outcome of the study, however unlikely this may be and provide a suitable reference to lead the reader to the data on gender effects of platelets.
Please rate the manuscript for methodological rigour
Very good
Please rate the quality of the presentation and structure of the manuscript
Very good
To what extent are the conclusions supported by the data?
Strongly support
Do you have any concerns of possible image manipulation, plagiarism or any other unethical practices?
No
Is there a potential financial or other conflict of interest between yourself and the author(s)?
No
If this manuscript involves human and/or animal work, have the subjects been treated in an ethical manner and the authors complied with the appropriate guidelines?
Yes
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The work presented is clear and the arguments well formed. This study would be a valuable contribution to the existing literature. This is a study that would be of interest to the field and community. The reviewers have highlighted minor concerns with the work presented. Please ensure that you address their comments.
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Comments to Author
This study by Lakerveld et al., reports experiments investigating the interaction with purified RSV (an important human pathogen) particles with freshly isolated human platelets as it has emerged recently that platelets play important roles in host response to - and control of - viral infections. This is a focused and convincing set of experiments, that use "gold-standard" approaches to address this important question alongside critical control experiments. The authors convincingly show evidence that RSV fails to classically activate platelets, despite binding and entering into them. I have a few suggestions that may help clarify areas in this manuscript prior to publication. My comments largely come from an interest in this area of research stemming from such an engaging article. Comments: Can the …
Comments to Author
This study by Lakerveld et al., reports experiments investigating the interaction with purified RSV (an important human pathogen) particles with freshly isolated human platelets as it has emerged recently that platelets play important roles in host response to - and control of - viral infections. This is a focused and convincing set of experiments, that use "gold-standard" approaches to address this important question alongside critical control experiments. The authors convincingly show evidence that RSV fails to classically activate platelets, despite binding and entering into them. I have a few suggestions that may help clarify areas in this manuscript prior to publication. My comments largely come from an interest in this area of research stemming from such an engaging article. Comments: Can the authors provide images of platelets during each of the treatments? The authors cannot comment on whether RSV replicate inside platelets using their current approach. While yes this system should only express GFP during replication, GFP detection by flow could be explained uptake of GFP-containing material during incubation. Yes this level of infection is much lower than would occur in highly permissive cell lines, it would be of great interest if RSV replicated at even low levels, therefore, the authors could: Use UV inactivated virus Bind with RSV and wash off and incubate and measure GFP Methods: did the authors use fresh pure particles or did they freeze/thaw before challenge? This reviewer disagrees on the MOI interpretation especially when comparing between virus stocks. As this work is largely independent of infectivity (maybe), a greater emphasis should be placed on particles/genome copies rather than MOI/pfu or tcid50. Can the authors comment on this, and modify any MOI- based conclusions? Are these platelets from a single donor? Especially thinking about the infection experiment. Can the authors provide a few lines on the passage history of their X strain, as it is not one that I had come across before and it is hard to find out information online. In particular, how many passages on what cells before cloning to cDNA? And then, how many passages and what cells before stocks used here? The authors should be cautious on their conclusions on the role of platelets in RSV pathology, as they only looked at a few inflammatory mediators, and more interestingly, this work raises the possibility that RSV actively suppresses platelet activation. Could this be looked at by co-incubation of RSV with positive control? It is interesting the detection of RSV associated chemokines. Can the authors speculate the reasons? Are they associated with virions as suggested, or could they come from additional non-viral structures like extracellular vesicles? Is there a chance that this indicates the sucrose purification did not work appropriately?
Please rate the manuscript for methodological rigour
Good
Please rate the quality of the presentation and structure of the manuscript
Very good
To what extent are the conclusions supported by the data?
Partially support
Do you have any concerns of possible image manipulation, plagiarism or any other unethical practices?
No
Is there a potential financial or other conflict of interest between yourself and the author(s)?
No
If this manuscript involves human and/or animal work, have the subjects been treated in an ethical manner and the authors complied with the appropriate guidelines?
Yes
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