PONE-D-21-36521SARS-CoV-2 spike-specific memory B cells express markers of durable immunity after non-severe COVID-19 but not after severe diseasePLOS ONE

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This work was supported by a COVID-19 pilot award from the UT Health Long School of Medicine (10009547 to E.M.B.). R.A.R. was supported by Translational Science Training award TL1 TR002647. Data were generated in the Flow Cytometry Shared Resource Facility, which is supported by UT Health, NIH-NCI P30 CA054174-20 (CTRC at UT Health) and UL1 TR001120 (CTSA grant). The following reagent was produced under HHSN272201400008C and obtained through BEI Resources, NIAID, NIH: Vector pCAGGS Containing the SARS-Related Coronavirus 2, Wuhan-Hu-1 spike glycoprotein receptor binding domain (RBD), NR-52309. SARS-CoV-2 spike HexaPro was a gift from Dr. Jason McLellan (Addgene plasmid #154754). 

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This work was supported by a COVID-19 pilot award from the UT Health Long School of Medicine (10009547 to E.M.B.). R.A.R. was supported by Translational Science Training award TL1 TR002647. Data were generated in the Flow Cytometry Shared Resource Facility, which is supported by UT Health, NIH-NCI P30 CA054174-20 (CTRC at UT Health) and UL1 TR001120 (CTSA grant). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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Additional Editor Comments:

The manuscript provides a detailed description of the B-cell responses SARS-CoV-2 after mild or severe COVID-19. As indicated by reviewer 1, the patient groups are not very large, but the work is very detailed and confirms and extends previous observations. I recommend a minor revision in which all of reviewer 1' s comments are addressed. In addition, I have 2 minor comments that need to be addressed:

- Please state how many spike and RBD specific cells were gated in each sample. Perhaps a median/range? This is important for the reader to assess whether there has been a true population (30 events), and how reliable subsetting of this population is in each sample.

- Please show pairwise comparisons of T-bet, FcRL5 and CD11c of total B cells in Fig 4 alongside this analysis on Ag-specific cells. Are these markers only different on antigen-specific cells? Or is there a general shift on all B cells? the first is purely related to the response. The latter (all B cells) could indicate effects of severe infection on the total immune system. It is very well possible that the 'cytokine storm' affects the B-cell compartment independent of antigen-specificity

Please make sure to discuss the outcomes of these 2 changes in the Discussion as well.

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Reviewers' comments:

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Reviewer #1: Partly

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2. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: Yes

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Reviewer #1: Yes

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Reviewer #1: Yes

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5. Review Comments to the Author

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Reviewer #1: The revised manuscript includes some minor edits to the text to clarify that the differences seen between non-severe and severe COVID-10 are subtle. However, the authors have not changed the title of the manuscript which states that spike-specific memory B cells do not express markers of durable immunity after severe disease.

Considering that the only significant differences between spike-specific memory B cells between the groups is a small decrease in T-bet and Fcrl5 expression in severe cases this title overstates the findings of the manuscript. Any conclusion about the duration of the spike-specific memory B cell response would require a longitudinal study of the severely infected patient cohort which the authors were unable to perform due to a lack of success in recruited recovered severely infected patients for a follow up blood draw. Therefore, it is not appropriate to imply in the title that the memory B cell response is not durable after severe infection. For this manuscript to be appropriate for publication the title should be revised to more accurately represent their findings that there is decreased expression of T-bet and Fcrl5 following severe infection rather than making a broader statement about the duration of immune memory.

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Reviewer #1: No

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SARS-CoV-2 spike-specific memory B cells express higher levels of T-bet and FcRL5 after non-severe COVID-19 as compared to severe disease

PONE-D-21-36521R1

Dear Dr. Bunnik,

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.

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Kind regards,

Menno C van Zelm, PhD

Academic Editor

PLOS ONE

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