PONE-D-22-17512Immunophenotype and antitumor activity of cytokine-induced killer cells from patients with hepatocellular carcinomaPLOS ONE

Dear Dr. Lin,

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process.

The manuscript has been evaluated by one reviewer, and his comments are available below.<o:p></o:p>

The reviewer has raised a number of concerns that need attention. He requests additional information on methodological aspects of the study such as the definition of CIK and the number of patients used. <o:p></o:p>

Could you please revise the manuscript to carefully address the concerns raised?<o:p></o:p>

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Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #1: Partly

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2. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: Yes

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3. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #1: Yes

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4. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #1: Yes

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Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: Major Points:

Please elaborate what is new finding in this paper?

Could you correlate expression of DNAM or NKG2D on K562 and HepG2 with the susceptibility to killing by CIK?

CIK from how many HCC patients were tested?

How are you defining CIK here; CD3+CD4+ T cells, CD3+CD8+ T cells or both?

Is CD56 expressed on CD3+CD4+ T cells or only on CD3+CD8+ T cells?

What was the %age of CD25highFoxP3+ T cells in expanded cells?

What would be the effects on CIK, if IL-15 is added to expand and induce CIK?

Why did not you investigate expression of other NK cell receptors like NKG2C, NKG2A, KIR, NKp46, etc?

Minor points:

Page 11: The results of other treatments for palliative purpose remain……(not remains).

Page 13: A renowned worldwide cancer treatment guideline has adopted………(provide ref).

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Reviewer #1: No

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PONE-D-22-17512R1Immunophenotype and antitumor activity of cytokine-induced killer cells from patients with hepatocellular carcinomaPLOS ONE

Dear Dr. Chang Lin,

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process. Kindly address comments raised by reviewer 2 and resubmit.

Please submit your revised manuscript by 20 December 2022. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org. When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file.

Please include the following items when submitting your revised manuscript:

• A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'.
• A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'.
• An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'.

If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter.

If applicable, we recommend that you deposit your laboratory protocols in protocols.io to enhance the reproducibility of your results. Protocols.io assigns your protocol its own identifier (DOI) so that it can be cited independently in the future. For instructions see: https://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols. Additionally, PLOS ONE offers an option for publishing peer-reviewed Lab Protocol articles, which describe protocols hosted on protocols.io. Read more information on sharing protocols at https://plos.org/protocols?utm_medium=editorial-email&utm_source=authorletters&utm_campaign=protocols.

We look forward to receiving your revised manuscript.

Kind regards,

Afsheen Raza, PhD

Academic Editor

PLOS ONE

Additional Editor Comments:

Kindly address the comments by reviewer 2 for manuscript revision

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Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation.

Reviewer #1: All comments have been addressed

Reviewer #2: (No Response)

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2. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #1: (No Response)

Reviewer #2: (No Response)

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3. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: (No Response)

Reviewer #2: (No Response)

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4. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #1: (No Response)

Reviewer #2: (No Response)

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5. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #1: (No Response)

Reviewer #2: (No Response)

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6. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: (No Response)

Reviewer #2: Manuscript ID: PONE-D-22-17512R1

Manuscript Title: Immunophenotype and antitumor activity of cytokine-induced killer cells from patients with hepatocellular carcinoma

The authors activated human cytokine-induced killer (CIK) cells ex-vivo and expanded from PBMCs from hepatocellular carcinoma (HCC) patients. They evaluated immunophenotyping and the ex-vivo killing ability of human CIK cells. Furthermore, they intravenously injected human CIK cells into NOD/SCID mice to evaluate the in vivo anticancer ability. They used human HCC cell line, J7, bearing NOD/SCID mice model in this study. This is an important subject and well written manuscript but there are several points should be addressed before its publication.

Non-obese-diabetic–severe combined immunodeficient mice (NOD–SCID) which were reported to lack innate immunity but are limited owing to a residual appearance of T and B cells and remaining natural killer (NK) cells (Cancer Res. 76, 6153–6158 (2016)). Recombination activating gene (RAG)-knockout mice overcome the residual appearance of adaptive immune cells but still show NK cells (Cancer Res. 76, 6153–6158 (2016)). NSG (NOD/SCID-IL2g-/−) strain mice, produced by the targeted mutation of the IL-2 receptor g-chain locus in a previously bred NOD–SCID strain, lack T cells, B cells, macrophages, NK cells and natural killer T cells (NKT) and have become an increasingly used platform for xenograft models (Cancer Res. 76, 6153–6158 (2016); Cancer Lett. 344, 13–19 (2014)). Therefore, we cannot tell the origin of antitumor effect in this study. The authors are better to use RAG-knockout mice or NSG (NOD/SCID-IL2g-/−) mice model in this study.

Another point to be addressed here is that human CIK cells could be affected by immunosuppressive cells (J Hepatol. 2019 Mar;70(3):449-457). Therefore, we need human immunosuppressive cells in murine HCC models to tell the exact anticancer effect of human CIK cells in vivo system.

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Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy.

Reviewer #1: No

Reviewer #2: No

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Attachments

Attachment

Submitted filename: PONE-D-22-17512R1.doc

Immunophenotype and antitumor activity of cytokine-induced killer cells from patients with hepatocellular carcinoma

PONE-D-22-17512R2

Dear Dr. Lin,

We’re pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements.

Within one week, you’ll receive an e-mail detailing the required amendments. When these have been addressed, you’ll receive a formal acceptance letter and your manuscript will be scheduled for publication.

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Kind regards,

Afsheen Raza, PhD

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

Reviewers' comments: