PONE-D-22-32175Quantitative proteomic analysis of extracellular vesicles in response to baculovirus infection of a Trichoplusia ni cell linePLOS ONE

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Reviewer #1: Partly

Reviewer #2: Yes

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Reviewer #1: Yes

Reviewer #2: Yes

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Reviewer #1: Yes

Reviewer #2: Yes

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Reviewer #1: Yes

Reviewer #2: Yes

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Reviewer #1: This paper presents for the first time an account of the isolation and proteomic characterisation of EVs from non-infected and baculovirus-infected Tni cells. The study is detailed and results presented suitably. I have just one concern that I think the authors should address prior to publication. EVs and baculovirus budded virus have very similar biophysical properties making it very hard to separate them completely. I can't find any recognition of this problem mentioned. What controls, or what limitations on the data and subsequent conclusions, did the authors put in place to ensure that the EVs isolated from baculovirus-infected cells were not contaminated by BV? Are the authors confident that the data from infected cell EVs was not at least partially attributable to the presence of BV? Did the authors demonstrate that their method to purify EVs avoided co-purification of low levels of BV? Controls may well have been put in place but I couldn't find any mention of these in the current version of the paper. If the authors can't be certain, the text should be modified to address this potential limitation.

Reviewer #2: In this study the authors isolated and characterized EVs from Tnms42 cells, a derivative of High Five cells. These studies are interesting to the field of baculovirus expression vector system (BEVS). The results are solid and with a good scientific writing skill. Two concers should be included in the discussion section:

1. In BEVS, the most popular used cells are Sf9, why the authors used the High Five cells?

2. There are studies employed the microarray or RNAseq to analyze the gene expression profile change after the baculovirus infection in,Sf9 or High Five cells or BmN cells (derived from Bombyx mori). I am interesting to know whether the proteins expression change in EVs reveled in this manuscript also revealed in the previous studies, like (1) Expression of baculovirus genes in permissive and nonpermissive cell lines Biochemical and Biophysical Research Communications Volume 323, Issue 2, 15 October 2004, Pages 599-614. (2) DNA microarrays of baculovirus genomes: differential expression of viral genes in two susceptible insect cell lines J. Yamagishi, R. Isobe, T. Takebuchi & H. Bando Archives of Virology volume 148, pages587–597 (2003). I suggest these studies should be included in the studies to check the consistency.

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Reviewer #1: No

Reviewer #2: Yes

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Quantitative proteomic analysis of extracellular vesicles in response to baculovirus infection of a Trichoplusia ni cell line

PONE-D-22-32175R1

Dear Dr. Grabherr,

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,

Jian Xu, Ph.D.

Academic Editor

PLOS ONE

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