Increased Otoferlin Expression in B Cells Is Associated with Muscle Weakness in Untreated Juvenile Dermatomyositis: A Pilot Study

Round 1

Reviewer 1 Report

The authors report an association between increased expression of Otoferlin in B cells and muscle weakness in untreated juvenile dermatomyositis, which may be a potential prognostic indicator. This is a pilot study, and it's interesting.

Possibly a syntax error on page 2, line 10.

Figure 1 is not clear enough.

Author Response

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Reviewer 2 Report

The authors described the relationship between otoferlin and disease activity in untreated children with JDM.

There was a significant increase in otoferlin expression in JDM children compared to controls. Otoferlin expression decreased significantly after 2-3 months of treatment. Higher otoferlin expression was also associated with increased disease activity. In addition, B cells and plasmablasts were the primary cells expressing otoferlin. 

The authors concluded that the results of this pilot study suggest that otoferlin expression is increased in JDM patients compared to healthy controls and is positively correlated with disease activity markers. 

This is an interesting study and the paper is well written. I have a few questions.

How about a longer introduction and discussion?

What percentage of the cases in this study met the EULAR/ACR 2017 idiopathic inflammatory myopathy classification criteria?

What are the kinetics of otoferrin in glucocorticoid-induced myopathy?

I would like to know more about the mechanism by which otoferrin contributes to muscle weakness in JDM patients. 

Author Response

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Reviewer 3 Report

Dear Authors,

Compact and clear presentation. However, I have some objections:

1) Introduction. Please, give more specific basis for your aim, - why otoferlin detection is so important and is it so, that its alone increase is characteristic for JDM...? Without concentration detection (ElISA, PCR) you cant prove the increased expression...

2) Materials and methods. 2.1 please give clear inclusion/exclusion criteria for the controls;

2.3 please, give reference for the immunohistochemistry method/protocol;

The same is requested for the positive cell counting method!

You placed an example for counting in the Suppl. File Figure 3, but I have some objections here: - how the died and defective cells were excluded from the counting? I can see also some artefacts in this picture, sorry, this is not acceptable for the serious presentation.  This inqualitative microphoto cant be used for the illustration! Please, change it (and also in next one remove artefacts, if you have such ones!) Next, - what magnification was used for the counting, give reference for the counting method, please!

Finally, where is description of positive and negative controls for the antibodies. Please, include this in the subsection.

Results. Disagree with the Figure 3. Not indicated the immunohistochemistry in the title, not showed the magnification. Not understandable the connection between the muscle and Tonsil CD19...! Please, re-arrange and correctly describe the microphotos, this is not done now!

Conclusions. Shorten and make them more precise. Remove, please, the 2nd sentence which is useless for the conclusions.

References. 27 is small number for serious presentation.. But what is worse, - 2 previous century references what is not good for nowadays manuscript. Please, remove them or replace with more modern ones!

Author Response

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Round 2

Reviewer 3 Report

Dear Authors,

I agree with all what you have corrected.

BUT, - an exception is SUPPL. figure sFig3. Sorry, I am morphologist and would never accept something like this! Even more, - you cant use your counting method for such a non-qualitative region. But there is a solution and let me give advice to you... There are a tissues where is really difficult to get nice pictures and some limitations persist for the usage of computer counting method, for sure, but you have to limit then the size of non-qualitative regions. I enclosed your picture where is indicated with red lines (by me) what you can simply to remove (cut away) and one region (right side beneath the upper cut-line) could be improved by the special image improvement Program. These changes will not affect your Program data, but just remove the unacceptable places... Of course, a little bit from "wrong" color will stay, but generally the picture will be acceptable for the all specialists. You know, we have no any right to demonstrate non-qualitative images for our readers. IJMS is very serious Journal, your paper is excellent, except this picture and I insist you to improve this picture maximally for those who will interested in your paper!

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