Journal: Molecular Therapy Oncology

Article Title: Repurposing the SSRI paroxetine increases lymphocyte mobilization and improves the efficacy of measles virus-based immunovirotherapy

doi: 10.1016/j.omton.2025.201109

Figure Lengend Snippet: MV-s-NAP-uPA + aPD-1 + aTIGIT + paroxetine combination increases S1P1 expression on BM lymphocytes in an orthotopic CT-2A/C57BL/6 mouse model (A) Schematic illustration of the treatment plan. (B) Median S1P1 staining in the BM lymphocytes 8 days post-CT-2A implantation. (C) Median S1P1 staining in the BM lymphocytes 13/14 days post-CT-2A implantation. (D) Median S1P1 staining in the CD4+ BM lymphocytes 13/14 days post-CT-2A implantation. (E) Median S1P1 staining in the CD8+ BM lymphocytes 13/14 days post-CT-2A implantation. (F) Median S1P1 staining in the NKT BM lymphocytes 13/14 days post-CT-2A implantation. The graphs represent the mean values obtained with flow cytometry analysis of the isolated tissue. Statistical analysis was conducted with a one-way ANOVA followed by Tukey’s multiple comparisons test, with 3–4 mice analyzed per group at each time point. p values less than 0.05 were considered significant.

Article Snippet: The antibodies used were the following: aPD-1 (anti-mPD-1-mIgG1e3, clone: RMP1-14, rat IgG2a,κ and features mouse IgG1e3 isotype-constant region; InvivoGen, San Diego, CA, CAT #mpd1-mab15-10) and aTIGIT (anti-mTIGIT-mIgG2a, clone: 10A7, hamster IgG2a and features the mIgG2a isotype-constant region; InvivoGen, San Diego, CA, CAT #mtigit-mab10-10).

Techniques: Expressing, Staining, Flow Cytometry, Isolation

Journal: Molecular Therapy Oncology

Article Title: Repurposing the SSRI paroxetine increases lymphocyte mobilization and improves the efficacy of measles virus-based immunovirotherapy

doi: 10.1016/j.omton.2025.201109

Figure Lengend Snippet: Paroxetine mobilizes lymphocytes following treatment with MV-s-NAP-uPA oncolytic virotherapy and immunovirotherapy in an orthotopic CT-2A/C57BL/6 mouse model (A) Schematic illustration of the treatment plan. (B) Percentage of circulating lymphocytes among all immune cells in PBMCs on days 13 and 18 post-CT-2A implantation. (C) Standardized counts of CD4+ and CD8+ single-positive PBMCs following MV-s-NAP-uPA + aPD-1 + aTIGIT immunovirotherapy with and without paroxetine on day 13/14 post-CT-2A implantation. (D) Standardized counts of CD4+ and CD8+ single-positive PBMCs following MV-s-NAP-uPA oncolytic virotherapy with and without paroxetine on day 13/14 post-CT-2A implantation. (E) Comparison of thymus weights in mice reaching a predetermined sacrifice endpoint, evaluating the effects of paroxetine vs. respective controls. For (B–D), the graphs represent the mean values obtained with flow cytometry analysis of the isolated tissue. Statistical analysis was conducted with one-way ANOVA followed by Tukey’s multiple comparisons test, with 3–4 mice analyzed per group at each time point for (B–D) and 11–26 mice analyzed per group for (E). p values less than 0.05 were considered significant.

Article Snippet: The antibodies used were the following: aPD-1 (anti-mPD-1-mIgG1e3, clone: RMP1-14, rat IgG2a,κ and features mouse IgG1e3 isotype-constant region; InvivoGen, San Diego, CA, CAT #mpd1-mab15-10) and aTIGIT (anti-mTIGIT-mIgG2a, clone: 10A7, hamster IgG2a and features the mIgG2a isotype-constant region; InvivoGen, San Diego, CA, CAT #mtigit-mab10-10).

Techniques: Comparison, Flow Cytometry, Isolation

Journal: Molecular Therapy Oncology

Article Title: Repurposing the SSRI paroxetine increases lymphocyte mobilization and improves the efficacy of measles virus-based immunovirotherapy

doi: 10.1016/j.omton.2025.201109

Figure Lengend Snippet: Paroxetine enhances lymphocyte activation in lymphoid organs and peripheral blood following treatment with MV-s-NAP-uPA + aPD-1 + aTIGIT immunovirotherapy in an orthotopic CT-2A/C57BL/6 mouse model (A) Schematic illustration of the treatment plan. (B) Percentage of CD69+ double-positive (CD4+CD8+) and single-positive helper T cells (CD4+) in the thymus of mice treated with immunovirotherapy with and without paroxetine on day 8 post-CT-2A implantation. (C–E) Percentage of CD25+/FOXP3− CD4+ cells in the thymus, spleen, and BM of mice treated with immunovirotherapy with and without paroxetine on day 8 post-CT-2A implantation. (F) Percentage of CD25+/FOXP3− CD8+ cells in the peripheral blood of mice treated with immunovirotherapy with and without paroxetine on days 8 and 13 post-CT-2A implantation. (G) Percentage of Tregs in the BM of mice treated with immunovirotherapy with and without paroxetine on day 18 post-CT-2A implantation. (H) Percentage of TIM-3+ lymphocytes in the thymus on day 18 post-CT-2A implantation. (I) Percentage of PD-1+ CD8+ cells in the spleen on day 18 post-CT-2A implantation. The graphs represent the mean values obtained with flow cytometry analysis of the isolated tissues. Statistical analysis was conducted with student two-tailed t tests with 3–4 mice analyzed per group at each time point. p values less than 0.05 were considered significant.

Article Snippet: The antibodies used were the following: aPD-1 (anti-mPD-1-mIgG1e3, clone: RMP1-14, rat IgG2a,κ and features mouse IgG1e3 isotype-constant region; InvivoGen, San Diego, CA, CAT #mpd1-mab15-10) and aTIGIT (anti-mTIGIT-mIgG2a, clone: 10A7, hamster IgG2a and features the mIgG2a isotype-constant region; InvivoGen, San Diego, CA, CAT #mtigit-mab10-10).

Techniques: Activation Assay, Flow Cytometry, Isolation, Two Tailed Test

Journal: Molecular Therapy Oncology

Article Title: Repurposing the SSRI paroxetine increases lymphocyte mobilization and improves the efficacy of measles virus-based immunovirotherapy

doi: 10.1016/j.omton.2025.201109

Figure Lengend Snippet: The effect of paroxetine on the survival in an orthotopic CT-2A/C57BL/6 mouse model treated with MV-s-NAP-uPA + aPD-1 + aTIGIT immunovirotherapy (A) Schematic illustration of the treatment plan in the pilot survival mouse experiment. (B) Kaplan-Meier plot representing the survival in the pilot study evaluating the effect of paroxetine and/or G-CSF in combination with immunovirotherapy (n = 4–8 mice per group). (C) Schematic illustration of our rechallenge experiment. (D) Kaplan-Meier plot representing the survival following rechallenge of the surviving mice from each group 180 days after the original tumor implantation with the same CT-2A cell line or a foreign melanoma B16-F10 cell line (n = 3–4 mice per group). Mice were rechallenged with intracranial implantation. (E) Schematic illustration of the treatment plan for the survival experiment presented in F. (F) Kaplan-Meier plot representing the survival following treatment with paroxetine in combination with oncolytic virotherapy, immunotherapy, and immunovirotherapy (n = 6–17 mice per group). p values less than 0.05 were considered significant.

Article Snippet: The antibodies used were the following: aPD-1 (anti-mPD-1-mIgG1e3, clone: RMP1-14, rat IgG2a,κ and features mouse IgG1e3 isotype-constant region; InvivoGen, San Diego, CA, CAT #mpd1-mab15-10) and aTIGIT (anti-mTIGIT-mIgG2a, clone: 10A7, hamster IgG2a and features the mIgG2a isotype-constant region; InvivoGen, San Diego, CA, CAT #mtigit-mab10-10).

Techniques: Tumor Implantation

Journal: Molecular Therapy Oncology

Article Title: Repurposing the SSRI paroxetine increases lymphocyte mobilization and improves the efficacy of measles virus-based immunovirotherapy

doi: 10.1016/j.omton.2025.201109

Figure Lengend Snippet: Combination therapy with MV-s-NAP-uPA + aPD-1 + aTIGIT + paroxetine does not increase toxicity in the orthotopic CT-2A/C57BL/6 mouse model (A) Mice from the survival studies were weighed every 3–4 days, and the percent change from baseline is presented longitudinally. Except for a brief post-operative weight loss, which was consistent across vehicle-treated mice and those receiving combinations based on MV-s-NAP-uPA, all treated mice, regardless of their treatment groups, gained weight at a comparable rate. (B) Cytokine analysis for IL-1β, IL-2, IL-4, IL-5, IL-6, IL-13, IFN-γ, IL-12p70, GM-CSF, TNF-α, and IL-18 was performed on plasma samples collected from mice on days 8, 13/14, and 18/19 post-CT-2A implantation. The graphs in (B) represent the mean values obtained from 3–4 mice per group at each time point.

Article Snippet: The antibodies used were the following: aPD-1 (anti-mPD-1-mIgG1e3, clone: RMP1-14, rat IgG2a,κ and features mouse IgG1e3 isotype-constant region; InvivoGen, San Diego, CA, CAT #mpd1-mab15-10) and aTIGIT (anti-mTIGIT-mIgG2a, clone: 10A7, hamster IgG2a and features the mIgG2a isotype-constant region; InvivoGen, San Diego, CA, CAT #mtigit-mab10-10).

Techniques: Clinical Proteomics

Journal: Advanced Science

Article Title: PD‐L1 on Tumor‐Derived Extracellular Vesicles Induces CD8 + T Cell Terminal Exhaustion and Mediates Anti‐PD‐1 Resistance in Head and Neck Squamous Cell Carcinoma

doi: 10.1002/advs.202516348

Figure Lengend Snippet: Differences in the number of tumor‐infiltrating exhausted T cells before and after aPD‐1 treatment in HNSCC. A) UMAP plot illustrating the distribution of tumor‐infiltrating CD8 + T cells from the GSE200996 (n = 25) and GSE195832 (n = 8) datasets, color‐coded by the three broad cell groups (C1, C2, and C3). The plot shows a comparison of pretreatment and posttreatment samples, highlighting the expression levels of key markers, such as GZMK , CXCL13 , TIM3 , CTLA4 , LAYN , and MKI67 , in CD8 + T cells. B) Comparison of the proportions of three distinct clusters of CD8 + T cells between pretreatment and posttreatment samples. C) Schematic diagram illustrating the mouse subcutaneous transplantation tumor model of HNSCC treated with IgG as a control or aPD‐1 treatment on days 6, 8, 10, and 12 after tumor implantation (n = 4 per group). Created in https://BioRender.com . D) Tumor growth curves of the mice in different treatment groups; the arrows represent the time points of drug administration. E) Proportion of tumor‐infiltrating CD8 + T‐cell subsets in mice in different treatment groups. F) Flow cytometry analyzing the infiltration ratios of PD‐1 + TCF‐1 − CD8 + T (Tex term ) cells, PD‐1 + TCF‐1 + CD8 + T (Tex prog ) cells, and PD‐1 + CD8 + T cells in response to IgG or aPD‐1 treatment. The data are presented as the mean ± SEM. ns, not significant, * P < 0.05, ** P < 0.01, two‐tailed Student's t test. Pre, pretreatment; Post, posttreatment; Tex term cells, terminally exhausted T cells; Tex prog cells, precursor exhausted T cells; prolif, proliferation; aPD‐1, anti‐PD‐1; i.p., intraperitoneal injection; FCM, flow cytometry.

Article Snippet: For in vivo antibody treatment, the mice received intraperitoneal injections of either 200 μg of the IgG1 isotype control (BioXCell, #BE0083) or aPD‐1 antibodies (BioXCell, #BE0146) every three days for two weeks.

Techniques: Comparison, Expressing, Transplantation Assay, Control, Tumor Implantation, Flow Cytometry, Two Tailed Test, Injection

Journal: Advanced Science

Article Title: PD‐L1 on Tumor‐Derived Extracellular Vesicles Induces CD8 + T Cell Terminal Exhaustion and Mediates Anti‐PD‐1 Resistance in Head and Neck Squamous Cell Carcinoma

doi: 10.1002/advs.202516348

Figure Lengend Snippet: TME infiltration of CD8 + T‐cell subsets in the aPD‐1 treatment responder group and non‐responder group was assessed. A) Summary of the procedure used to evaluate CD8 + T‐cell subset infiltration in the TME of 40 patients receiving immune checkpoint inhibitor (ICI) treatment, and the RECIST 1.1 criteria for assessing immunotherapy efficacy. Created in https://BioRender.com . B) Sankey diagram illustrating patient transfer during immunotherapy, infiltration of PD‐1 + TCF‐1 − CD8 + T (Tex term ) cells within the stroma or tumor, and survival status in the clinical cohort. C) ROC analysis of the ability of Tex term cell infiltration levels to predict the efficacy of immunotherapy. D) Multiplex immunofluorescence staining of tumor tissues from HNSCC patients was performed to detect CD8 (yellow), PD‐1 (green), TCF‐1 (red), Pan‐CK (light green), and DAPI (blue) expression, with a scale bar of 100 µm. The infiltration of different subsets of CD8 + T cells in the stroma, tumor, and TME between the responder (CR+PR) groups (n = 26) and the non‐responder (SD+PD) groups (n = 14) was statistically analyzed. E) Flow chart of the in vivo animal experiments. Created in https://BioRender.com . F) Tumor volume curves of SCC7 tumor‐bearing C3HeN mice treated with IgG as a control (n = 7) or aPD‐1 (n = 15) twice a week (arrows represent time points of administration). The mice in the aPD‐1 treatment group were divided into a responder group and a non‐responder group according to tumor size. Significance was calculated by two‐way ANOVA. G) The proportions of CD8 + T‐cell subsets in the tumor microenvironment of mice in the aPD‐1 treatment responder group and non‐responder group were analyzed by flow cytometry. The data are presented as the mean ± SEM. ** P < P0.01, *** P < 0.001; two‐tailed Student's t test. PR, partial response; CR, complete response; PD, progressive disease; SD, stable disease; Tex term cells, terminally exhausted T cells; Tex prog cells, precursor exhausted T cells; AUC, area under the curve; TME, tumor microenvironment; R, responder; NR, non‐responder; i.p., intraperitoneal injection; wt, wild type; aPD‐1, anti‐PD‐1.

Article Snippet: For in vivo antibody treatment, the mice received intraperitoneal injections of either 200 μg of the IgG1 isotype control (BioXCell, #BE0083) or aPD‐1 antibodies (BioXCell, #BE0146) every three days for two weeks.

Techniques: Multiplex Assay, Immunofluorescence, Staining, Expressing, In Vivo, Control, Flow Cytometry, Two Tailed Test, Injection