Roles for ADAM17 in TNF-R1 Mediated Cell Death and Survival in Human U937 and Jurkat Cells

Round 1

Reviewer 1 Report

I have reviewed the manuscript and consider it well written and very interesting. It is suitable for publication as it is. Best regards.

Author Response

We would like to thank the reviewer for the kind words and appreciation of our work.

Reviewer 2 Report

In the manuscript, entitled “Roles for ADAM17 in TNF-R1 mediated cell death and survival in human U937 and Jurkat cells”, the authors demonstrated the role of the enzyme ADAM17 in death receptor signaling in human hematopoietic cells using U937 and Jurkat cell lines. They showed that absence of ADAM17 increases TNF-mediated cell death. They also showed that cell death triggered via DR3 is enhanced in U937 cells lacking ADAM17. They concluded that ADAM17 is a key molecule in fining tuning death receptor signaling. This study is interesting and will provide a better understanding of cell fate decisions made via receptors of the TNF-R1 superfamily. However, there are a few points are required to be addressed by the authors:

• The title of the article does not precisely describe the major finding(s) of the study so I think it should be modified.
• P-value should be calculated for all experimental results performed in replicates to evaluate the statistical significance of differences between groups.
• In the subheading entitled “Analysis of Cell death induction upon pharmacological ADAM17 inhibition”, the authors correctly describe the effect of the ADAM17 inhibitor TAPI or the ADAM10 inhibitor GI254023X on the cell death of U937 and Jurkat cell lines. However, the results of the mEF cells should be moved to the subheading entitled “Genetic ADAM17 deficiency has differential effects on TNF induced cell death in U937 and 267 Jurkat cells” because ADAM17 in these cells is genetically inhibited (not pharmacologically). Also, the results of the mEF cells should be moved from figure 1 to 4.
• The authors should add in the perspective part of the discussion that their findings should be tested in human primary cells such as peripheral blood monocytes and lymphocytes.

Minor Points:

• Line 22: “increase” should be changed to “increases”
• Line 57: ADAM7 should be changed to ADAM17
• Line 209: Fig.2C should be changed to Fig.2B
• Figure 2: Please remove THP-1 cells from the legend

 

Author Response

We would like to thank the reviewer for the appreciation of our work and the helpful comments.

- The typos in lines 22, 57, 209 and in Fig 2 have been corrected accordingly.

- A paragraph stating that testing in primary human cells should be performed has been added in line 462.

- In Figure 1, mEF experiments have been removed and shifted to Figure 4. Text has been adapted accordingly

- p-values have been added where applicable and variance allowed.

- As our observations have several aspects, we decided to leave the article title as it is. Otherwise, the title would become too complex in our eyes.

 

We hope we were able to address all raised issues adequately.