SARS-CoV-2 Structural Proteins Modulated Blood-Testis Barrier-Related Proteins through Autophagy in the Primary Sertoli Cells

Round 1

Reviewer 1 Report

Comments to Authors 

 

            This study showed that ectopic expression of viral E (envelope protein) and M (membrane protein) induced the expressions of ZO-1 and claudin11, promoted the formation of autophagosomes, and inhibited autophagy flux.

          Authors are kindly requested to emphasize the current concepts about these issues in the context of recent knowledge and the available literature. This articles should be quoted in the References list.

References

1.      SARS-CoV-2 Harnesses Host Translational Shutoff and Autophagy To Optimize Virus Yields: the Role of the Envelope (E) Protein. Microbiol Spectr. 2023;11(1):e0370722. doi:10.1128/spectrum.03707-22.

2.      Characterization of the induction kinetics and antiviral functions of IRF1, ISG15 and ISG20 in cells infected with gammacoronavirus avian infectious bronchitis virus. Virology. 2023;582:114-127. doi:10.1016/j.virol.2023.03.017.

Minor editing of English language required

Author Response

1 Point:This study showed that ectopic expression of viral E (envelope protein) and M (membrane protein) induced the expressions of ZO-1 and claudin11, promoted the formation of autophagosomes, and inhibited autophagy flux.

          Authors are kindly requested to emphasize the current concepts about these issues in the context of recent knowledge and the available literature. This articles should be quoted in the References list.

     SARS-CoV-2 Harnesses Host Translational Shutoff and Autophagy To Optimize Virus Yields: the Role of the Envelope (E) Protein. Microbiol Spectr. 2023;11(1):e0370722. doi:10.1128/spectrum.03707-22.

     Characterization of the induction kinetics and antiviral functions of IRF1, ISG15 and ISG20 in cells infected with gammacoronavirus avian infectious bronchitis virus. Virology. 2023;582:114-127. doi:10.1016/j.virol.2023.03.017.

Response: Thank you for your advice and information. The article“SARS-CoV-2 Harnesses Host Translational Shutoff and Autophagy to Optimize Virus Yields: the Role of the Envelope (E) Protein” had been added in the introduction, listed as references [22]. We have read the article “Characterization of the induction kinetics and antiviral functions of IRF1, ISG15 and ISG20 in cells infected with gammacoronavirus avian infectious bronchitis virus” carefully, and agreed that this article is not closely related to our research report, so we finally decided not to cite this article. Finally, we cited the new article you recommended and re-edited all references to the manuscript.

2. Point:Comments on the Quality of English Language

Minor editing of English language required

Response: Thank you for your warm comment. We carefully revised the manuscript again, and invited a native English professional to improve the language and grammar of the full text.

 

 

Reviewer 2 Report

As the SARS-CoV-2 pandemic progressed, the virus was shown to affect multiple organ systems, some in very profound ways.  Since angiotensin converting enzyme (ACE2), the receptor for the virus, is expressed at high levels in testicles, the male reproductive system is no exception.  A moderate COVID-19 infection has been shown to negatively affect spermatogenesis with a more severe infection resulting in changes in the blood-testis barrier (BTB) and in testicular cells themselves with a concomitant increase in immune factors.  In this study, the effects of the individual viral proteins on BTB-related proteins and the levels of immune factors, in addition to the formation and degradation of autophagosomes in Sertoli cells are investigated through ectopically expressed individual SARS-CoV-2 viral proteins.  The study convincingly demonstrates that the viral E, M, N and S proteins modulate the expression of BTB-related proteins via an effect on autophagy.  Although the study is well-performed and the conclusions appear to be supported by the data, there are a number of problems with the manuscript that must be addressed prior to publication. 

 

First and foremost, the manuscript is very poorly written.  There are a multitude of examples of poor usage of the English language, grammar and spelling.  Just one example is the frequent use of the term lever for level.  Another is the use of “infected” instead of “transfected” (line 192).  These are just a few examples.  The manuscript requires very significant editorial attention.

 

The authors should be more careful with the selective interpretation of the data.  There are instances in which they attribute effects to one or more proteins, but not others that have the same effects.

 

Specific points

The reader needs to be told more about Sertoli cells when they are first mentioned in the manuscript (line 56).

 

Figure 2 A and B: The effects of the M protein on ZO-1 in Figures A-a and B-b do not agree.

 

Line 206: Contrary to the statement made here, there are significant changes in catenin with M and S (Fig. 2A,e).  Also, there is disagreement in the effect of M and S on catenin in Fig. A, e and B, f.

 

Lines 217-8: N does not induce the expression of TGF and IL-1, as stated here.. Do the authors mean to cite the E protein here?

 

Lines 283-4: While it is true that M and N enhance CX43 expression, isn’t this also true of S?  This is just one example of selective interpretation of the data to suit the point the authors want to make.  There are numerous others.

Very poor usage of the English language

Author Response

Please see the attachment.

Author Response File: Author Response.pdf