Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

Good morning for the authors,

Analyzing the manuscript (Article) with ID ijms-3130057-peer-review-v1, entitled "Cystatin F Depletion in Mycobacterium tuberculosis-infected Macrophages Improves Cathepsin C/Granzyme B-driven Cytotoxic Effects on HIV-infected Cells During Coinfection" for possible publication in the International Journal of Molecular Sciences, Section: Molecular Pathology, Diagnostics, and Therapeutics, Special Issue: The Role of Lysosomal Proteases in Cancer and Infectious Diseases

I consider that:

1.                  The authors proposed a much-discussed topic in today's medical scientific world: Coinfection Mycobacterium tuberculosis and HIV.

2.                  This article follows all the specific instructions of the journal presented in aims and scope, instructions for authors, and other information about the International Journal of Molecular Sciences.

3.                  In Chapter Introduction: The authors present relevant and important medical information for the chosen topic: The Coinfection Mycobacterium tuberculosis and HIV.

4.                  In Chapter Results: The results of this study are well presented and easy to understand in the text of the manuscript:

-                     Sub-chapter 2.1. Experimental Conditions for Transfection and Infection Produce Comparable Low Levels of Cell Death and Figure 1 (a-b) are suggestive of the study conducted and very clearly presented;

-                     Sub-chapter 2.2. Decreased CstF Levels from Mtb-infected Macrophages are Correlated with Increased Enzymatic Activity of Cathepsin C in Lymphocytes During Coinfection and Figures 2 and 3 (a-b): are suggestive of the study carried out;

-                     This study was carried out in dynamics, and the results presented by the authors in the two sub-chapters 2.3 CstF Depletion is Correlated with Increased Cathepsin C-Granzyme B Driven Cytotoxic Effects, and 2.4. CstF Depletion Improves Cathepsin C/Granzyme B-Driven Reduction of Viral Replication During Mtb-HIV Coinfection, Figures 4 – 6 and ijms-3130057-original-images are clear.

5.                  In Chapter Discussion: According to the bibliography, the authors compared all the results obtained in this study with the results of other studies and the assessments of different authors. Figure 7 is very good!

6.                  In Chapter Materials and Methods: The authors made a special effort from the beginning of the study and presented very clearly:

-                     The human blood used in this study was obtained from a renowned institute. All standard work protocols for the processing of biological blood samples were followed (Sub-chapter 4.1. Cell Isolation and Culture Conditions);

-                     M. tuberculosis used in the study was obtained from an accredited and prestigious laboratory. The working protocols for the realization of co-cultures are the standard ones and validated by other authors in their studies (Sub-chapters 4.2. Bacterial Cultures and HIV Isolates and 4.3. Macrophage Infection). The authors specified that all experimental procedures using Mtb and HIV were performed in the biosafety level 3 laboratory!

-                     The authors used modern technologies (Sub-chapters: 4.6 Western blotting; 4.8. HIV Quantification - Colorimetric enzyme immunoassay; 4.9. Cell death and Viability assays using Flow Cytometry) standardized work protocols, high-performance medical equipment, and certified human kits. The kits, culture media, and reagent kits are supplied by specialized and prestigious companies.

-                     For the statistical analysis of the data, the authors used high-performance software: GraphPad Prism 9. The statistical parameters used by the authors are those specific to medical scientific studies (t-test and p-value < 0.05; Sub-chapter: 4.10. Statistical Analysis).

7.                  They had the approval of the Ethics Committee of the Hospital/Institute, thus proving high professional ethics.

8.                  The authors also received funding for this study, demonstrating professionalism and medical scientific relevance (presented in Funding).

9.                  All authors have made an equitable contribution to the study.

10.              The bibliography chosen by the authors corresponds to the requirements and refers to the subject of the article.

In conclusion:

I ACCEPT for the publication of this article in the International Journal of Molecular Sciences (IJMS, MDPI, ISSN:1422-0067, IF=4,9).

Congratulations to all authors for this article! Parabéns a todos os autores por este artigo! 

Comments for author File: Comments.pdf

Author Response

We thanks the reviewers work on review this manuscript and the positive comments.

I understand that the reviewers accepted the manuscript in the present form.

Reviewer 2 Report

Comments and Suggestions for Authors

Dear Authors,

thank you very much for your well-written manuscript. The subject of your study is highly important, as Mycobacterium tuberculosis (Mtb) and human immunodeficiency virus (HIV) are syndemic interaction pathogens and synergize an accelerated progression to tuberculosis (TB) and to acquired immune deficiency syndrome (AIDS) during coinfection. TB remains a significant public health concern, with one of the contributing factors being the synergistic effect of the coinfection with HIV. So, developing new therapeutic targets for co-infected patients may dramatically influence the global disease burden and reduce their impact on morbidity and mortality worldwide.  Your manuscript fits the purpose and scientific subject of the International Journal of Molecular Sciences and particularly of the special issue: “The role of Lysosomal Proteases in Cancer and Infectious Diseases”. English language is fine and no grammatical or structural issues were detected. Your experiments have been described in a reproducible way and the graphical presentation of your results can be easily interpreted. Your results suggest the existence of an evasion mechanism that enables early HIV replication during coinfection with Mtb at the interface environment through the axis CstF/ cathepsin C/ granzyme B and this knowledge can be crucial for developing new therapeutic approaches to control both pathogens based on the manipulation of CstF. Your results may contribute to better understanding the achievements made so far and provide impulse for more research in the future. I have no questions and queries, pertaining to your manuscript.

Best Regards

Author Response

We thanks the reviewers work on review this manuscript and the positive comments.

I understand that the reviewers accepted the manuscript in the present form.