Soluble CD14 and Lipopolysaccharide-Binding Protein Are Not Superior to Soluble CD25 as Biomarkers for Sarcoidosis

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

Dear Authors 

This study evaluate sCD14 and LBP as biomarkers of sarcoidosis and compare with established markers such as sCD25.Besides you investigated  these biomarkers according to  specific organ involvement. Sarcoidosis is an important disease and specific biomarkers are needed. This topic is of scientific interest. sCD14 and LBP which are investigated in this study have important role for inflammation. Comparing them with the establish marker increased the power of the study. The research question original and well defined. The manuscript clear scientifically, relevant to the field, english language appropriate and understandable and presented in a well-structured manner. The ethics and data availability statements are adequate The manuscript’s experimental design is suitable for testing the hypothesis and its results can be reproducible. The data were interpreted easy to understand  and appropriately and consistently throughout the manuscript. The conclusions are consistent with the evidence and arguments presented. I would like to emphasize a few points and make suggestions

• Purpose of study was written as ‘aim of the present study was to evaluate the diagnostic performance of serum sCD14 and LBP as biomarkers of sarcoidosis, in comparison with established markers  such as sCD25 and ACE, in a cohort of well-characterized sarcoidosis patients. Additionally, we investigated whether these biomarkers were associated with specific organ involvement and examined their longitudinal changes during immunosuppressive therapy
•  But ACE was not included in correlation analyses. Besides Analysis of outcomes after immunosuppressive therapy was not presented in the study.
• I suggest that  ‘’ aim of the present study was to evaluate the diagnostic performance of serum sCD14 and LBP as biomarkers of sarcoidosis, in comparison with established markers  such as sCD25, in a cohort of well-characterized sarcoidosis patients. Additionally, we investigated whether these biomarkers were associated with specific organ involvement

• Exclusion criteria are specified only as autoimmune and autoinflammatory diseases. Other conditions affecting these markers  (acute, chronic infections, dysbiosis, liver and kidney diseases, metabolic diseases, malignancy are not added. It is recommended to write the exclusion criteria in detail

 

• No laboratory data was included. Acute phase reactants, liver, kidney function tests, calcium, hemogram parameters are recommended to be added if available
•  
• It would be better to perform additional analysis for markers according to acute fhase reactants to be able to say whether they are inflammation inducator.

 

• Prior Studies have shown sCD25 is consistently elevated and strongly associated with sarcoidosis activity.Activity score was not included in this study. It would be better to perform additional analysis for markers according to the disease activity score

 

• Line 359: The authors wrote ‘ Importantly, all four biomarkers (sCD25, sCD14, LBP and ACE) significantly decreased under immunosuppressive therapy during follow-up, indicating sensitivity to treatment this explanation. ‘The analysis details of this were not presented in the study. It is recommended to add this analysis and compare the change in markers according to treatments.For example ,Which is more specific in evaluating treatment response, which treatment affected which parameter?

 

• It is recommended to avoid repeating the results in the discussion, to give more examples to similar studies and to expand the discussion section by emphasizing their strengths and weaknesses according to them

 

• It is recommended to shorten the conclusion and write 1-2 sentences in the form of a summary emphasizing the important points

 

• New references are rarely included. And several references have minor punctuation errors. It is recommended to include newly references and arrange references.

 

• There are a few minor missspelling suggested to be corrected. 198,201 line font size is different
• In Table 1, it is recommended to add (n) after ACE use
• End of the table 2:  Number of organs involved 3 [2–5]. It is recommended to write more clearly what is explained here
• It is recommended to write the explanation of words which the abbreviation is used in the tables (such as LBP, sCD14 and sCD25,...) these under the table

 Best Regards

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

This manuscript evaluates the diagnostic performance of serum biomarkers in sarcoidosis, comparing sCD14 and LBP with established markers such as sCD25 and ACE. The study addresses an important clinical challenge because reliable circulating biomarkers for sarcoidosis diagnosis remain limited. While the study design is generally appropriate, before the manuscript can be considered for publication following justification and revision are requested.

1. The rationale for selecting sCD14 and LBP as candidate biomarkers should be more clearly explained. Although identified through extracellular vesicle proteomics, the biological link between these molecules and granulomatous inflammation should be elaborated.
2. The sample size (46 cases and 46 controls) is relatively small for biomarker validation studies. Please provide a power calculation or justification for the cohort size.
3. Clarify inclusion and exclusion criteria for sarcoidosis patients. Specify diagnostic confirmation methods, such as histopathological evidence of non-caseating granulomas and exclusion of infectious causes.
4. The ELISA methods used for biomarker quantification should include information about assay kits, sensitivity, intra-assay variability, and calibration procedures.
5. The role of body mass index as a covariate requires further justification. Explain the biological rationale for including BMI in the regression model.
6. The stepwise AIC model selection process should be described in detail, including variables initially included in the model.
7. The longitudinal analysis in 32 treated patients is interesting but requires more methodological detail, including treatment regimens and follow-up duration.
8. The decrease in biomarker levels after therapy should be interpreted cautiously, as this may reflect general inflammation reduction rather than disease-specific activity.
9. Discuss why LBP and sCD14 did not improve diagnostic performance despite prior proteomic evidence.
10. A comparative table summarizing performance characteristics of the evaluated biomarkers would improve clarity.
11. The discussion should compare findings with previous studies investigating sarcoidosis biomarkers, including the established role of Angiotensin-converting enzyme and Soluble interleukin-2 receptor alpha chain.
12. Address potential confounders such as infections, obesity, or metabolic syndrome that could influence sCD14 and LBP levels.
13. The limitations section should explicitly mention the small sample size, single-center design, and lack of external validation.
14. Strengthen the conclusion by emphasizing the need for multicenter validation studies and integration of immunometabolic biomarkers.
15. Refer this recent studies include in literate: https://doi.org/10.1007/s10753-023-01911-5, https://doi.org/10.36922/bh.3515, ttps://doi.org/10.1016/j.ajpath.2025.06.005

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Round 2

Reviewer 2 Report

Comments and Suggestions for Authors

No more comments.