-->PONE-D-26-00663-->-->Gene Expression Profiling of Recipient Immune Cells Induced by 7 × 19 CAR-T Cell Dosing in a Syngeneic Mouse Model-->-->PLOS One

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Reviewer #1: Partly

Reviewer #2: Yes

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Reviewer #2: Yes

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Reviewer #1: Yes

Reviewer #2: Yes

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Reviewer #1: The manuscript titled Gene Expression Profiling of Recipient Immune Cells Induced by 7 × 19 CAR-T Cell Dosing in a Syngeneic Mouse Model” by Yo Muraki et al. discusses the underlying mechanism of action of 7 x 19 CAR-T cells reported earlier. Earlier, the 7 x 19 CAR-T cells had shown the dependency on resident immune cells for their activity. Therefore, the paper focuses on the cellular composition and gene expression patterns of resident immune cells during the early tumour-induction phase in solid tumours, in the presence of 7 x 19 CAR-T cells to gain insights into the possible mechanism of action.

The experimental approach is good, but the data analysis and figure presentation may need further refinement. A more stringent criterion, like at least a 2-fold change, is suggested to be used for differentially expressed genes. The manuscript needs further refinement as suggested in the reviewer's report.

Reviewer #2: 1. In Figure 1, the authors show improved tumor reduction with 7×19 CAR-T cells compared to conventional CAR-T cells. However, no survival analysis is provided. Without survival data, it is difficult to assess the long-term therapeutic benefit of this treatment.

2. In Figure 2, the authors report increased percentages of CD8+ T cells and macrophages in the tumor microenvironment. However, only percentage data are presented, and absolute cell counts are not shown. This makes it unclear whether there is a true increase in immune cell numbers or only a relative change.

3. The study reports upregulation of multiple chemokines and immune-related pathways, but these findings are based only on transcriptomic analysis. No protein-level validation (such as ELISA) or functional assays are provided to confirm these observations.

4. The individual roles of IL-7 and CCL19 are not separately analyzed. Since both are expressed together in the CAR-T construct, it is unclear which factor is primarily responsible for the observed immunological effects.

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Reviewer #1: Yes: Khalid Hussain Bhat, PhD. SKUAST-K, India.

Reviewer #2: Yes: Apoorva Rajput

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Attachments

Attachment

Submitted filename: PONE-D-26-00663 Comments_Khalid.docx

Attachment

Submitted filename: Review 1.docx

Gene Expression Profiling of Recipient Immune Cells Induced by 7 × 19 CAR-T Cell Dosing in a Syngeneic Mouse Model

PONE-D-26-00663R1

Dear Dr. Chihiro Take,

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.

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Academic Editor

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