Journal: Science Advances

Article Title: Messenger RNA delivery of a dual CD25/CD122 affinity-tuned IL-2 variant prolongs exposure and potentiates antitumor T cell immunity

doi: 10.1126/sciadv.aeh9215

Figure Lengend Snippet: ( A to D ) MC38 tumor-bearing C57BL/6 mice ( n = 7 to 8) were treated with Alb–IL-2var RNA-LNP (Alb–IL-2var) starting on day 18 post–tumor inoculation (average tumor volume, 165 mm 3 ). Alb RNA-LNP served as a control. (A) Experimental design. (B) Flow cytometry analysis of tumor-infiltrating total CD8 + T cells, natural killer (NK) cells, and T reg cells. (C) Flow cytometry analysis of neoantigen-specific CD8 + T cells. (D) TCF1 + PD-1 + neoantigen-specific CD8 + T cells, as determined by flow cytometry. ( E ) MC38 tumor-bearing C57BL/6 mice ( n = 5 to 7) were treated with Alb–IL-2var RNA-LNP (Alb–IL-2var) on days 14 and 21 post–tumor inoculation (average tumor volume, 86 mm 3 ). Alb RNA-LNP served as a control. TOX + PD-1 + cells among tumor-infiltrating CD8 + T cells (left) and Rpl18-specific CD8 + T cells (right), as determined by flow cytometry. (B to E) Horizontal dotted lines represent the means of the control group. ( F ) Survival and individual tumor growth curves of MC38 tumor-bearing C57BL/6 mice ( n = 14) treated with Alb–IL-2var RNA-LNP (Alb–IL-2var), and/or anti–PD-L1 antibody (αPD-L1) starting on day 17 post–tumor inoculation (average tumor volume, 73 mm 3 ). Alb RNA-LNP and isotype antibody served as controls. Vertical dotted lines represent treatments. ( G ) Survival and individual tumor growth curves of MC38 tumor-bearing C57BL/6 mice ( n = 17) treated with LRT, Alb–IL-2var RNA-LNP (Alb–IL-2var), and αPD-L1. LRT began on day 19 post–tumor inoculation (average tumor volume, 90 mm 3 ), whereas Alb–IL-2var RNA-LNP and/or αPD-L1 were administered on day 26, when tumors exceeded 200 mm 3 . Vertical dashed line represents LRT, and vertical dotted lines represent αPD-L1/Alb–IL-2var RNA-LNP treatments. Alb RNA-LNP and isotype antibody served as controls. Student’s unpaired t test (A to E) and log-rank test (F and G, left). Single outliers as identified by Grubb’s test were removed (B). Not significant, P > 0.05; * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001; **** P ≤ 0.0001.

Article Snippet: MC38 tumor-bearing mice were treated with 3 μg of RiboCytokine (Alb–IL-2var RNA-LNP; Alb RNA-LNP as control) in PBS through intravenous injection on days 17, 24, 32, and 39 after tumor inoculation and intraperitoneally injected with 100 μg of αPD-L1 (InvivoGen, hpdl1-mab9) or isotype control antibodies (day 17: 200 μg antibody) on the same days as RiboCytokine treatments.

Techniques: Control, Flow Cytometry

Journal: Science Advances

Article Title: Messenger RNA delivery of a dual CD25/CD122 affinity-tuned IL-2 variant prolongs exposure and potentiates antitumor T cell immunity

doi: 10.1126/sciadv.aeh9215

Figure Lengend Snippet: ( A ) Survival and individual tumor growth curves of TC-1 tumor-bearing C57BL/6 mice ( n = 16) treated with HPV16 E7 RNA-LPX vaccine (vaccine), anti–PD-L1 antibody (αPD-L1), and Alb–IL-2var RNA-LNP (Alb–IL-2var). Vaccination began on day 13 post–tumor inoculation (average tumor volume, 140 mm 3 ), whereas Alb–IL-2var RNA-LNP and/or αPD-L1 were administered on day 18, when tumors exceeded 350 mm 3 . Alb RNA-LNP, irrelevant RNA-LPX, and isotype antibody served as controls. Vertical dashed lines represent vaccine treatment, and vertical dotted lines represent αPD-L1 or Alb–IL-2var RNA-LNP treatments. ( B to D ) TC-1 tumor-bearing C57BL/6 mice ( n = 7 to 10) were treated with HPV16 E7 RNA-LPX vaccine (vaccine), anti–PD-L1 antibody (αPD-L), and/or Alb–IL-2var RNA-LNP (Alb–IL-2var) starting on day 13 post–tumor inoculation (average tumor volume, 139 mm 3 ). Alb RNA-LNP, irrelevant RNA-LPX, and isotype antibody served as controls. (B) Experimental design. (C) Tumor-infiltrating total- and HPV16 E7-specific CD8 + T cells and T reg cells as well as the HPV16 E7-specific CD8 + T–to–T reg cell ratio as determined by flow cytometry. (D) Tumor-infiltrating NK cells, cDC1, cDC2, and M1 macrophages as determined by flow cytometry. Horizontal dotted lines represent the means of the control group. Log-rank test (A, left) and Kruskal-Wallis test with Dunn’s multiple comparison test (B to D). Not significant (n.s.), P > 0.05; * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001; **** P ≤ 0.0001.

Article Snippet: MC38 tumor-bearing mice were treated with 3 μg of RiboCytokine (Alb–IL-2var RNA-LNP; Alb RNA-LNP as control) in PBS through intravenous injection on days 17, 24, 32, and 39 after tumor inoculation and intraperitoneally injected with 100 μg of αPD-L1 (InvivoGen, hpdl1-mab9) or isotype control antibodies (day 17: 200 μg antibody) on the same days as RiboCytokine treatments.

Techniques: Flow Cytometry, Control, Comparison

Journal: bioRxiv

Article Title: Microbiome-derived hydroxyphenyl propanoates enhance antitumour immunity by potentiating gasdermin D activity in tumour-associated myeloid cells

doi: 10.64898/2026.04.23.720410

Figure Lengend Snippet: a, IL-1β activity in the supernatant of THP1-WT vs. - GSDMD -KO cells treated with LPS±HPPs for 16 hours, measured using HEK-Blue™ IL-1β reporter cells (three combined experiments, error bars represent standard error, one-way ANOVA with Bonferroni test); b, LDH release from THP1-WT vs. – GSDMD -KO cells treated with LPS±HPP for 16 hours (three combined experiments, one-way ANOVA with Bonferroni test); c-d, IL-1β activity in the supernatant of THP1-WT vs. - GSDMD -KO cells © and human PBMCs (d) treated with LPS and NG±HPPs for 16 hours. measured using HEK-Blue™ IL-1β reporter cells (three combined experiments, error bars represent standard error, one-way ANOVA with Bonferroni test); e-f, LDH release from THP1-WT vs. – GSDMD -KO cells € and human PBMCs (f) treated with LPS and NG±HPPs for 16 hours (three combined experiments, one-way ANOVA with Bonferroni test).

Article Snippet: To neutralize IL-1 receptor signalling, the following antibodies were used: anti-human IL-1α (1 μg/mL; clone 7D4; mabg-hil1a-3; InvivoGen), anti-human IL-1β (1 μg/mL; clone 4H5; mabg-hil1b-3; InvivoGen) and IgG1 isotype control (1 μg/mL; clone T8E5; mabg1-ctrlm; InvivoGen).

Techniques: Activity Assay

Journal: bioRxiv

Article Title: Microbiome-derived hydroxyphenyl propanoates enhance antitumour immunity by potentiating gasdermin D activity in tumour-associated myeloid cells

doi: 10.64898/2026.04.23.720410

Figure Lengend Snippet: a-b, tumour growth kinetics and OS for orthotopic M3-9-M OVA tumours in male WT or Gsdmd -KO mice receiving vehicle or 3,2-HPP treatments from day 1 post cell-implantation (n =8/group; two combined experiments, two-way ANOVA test for growth kinetics, Log-Rank test for OS); c, [IL-1β] in interstitial fluid harvested from 18-day old M3-9-M OVA tumours grown in male WT or Gsdmd -KO mice receiving vehicle or 3,2-HPP treatment from day 1 post-cell implantation (two combined experiments, one-way ANOVA with Bonferroni multiple comparison test); d, GSDMD peptides quantified from a publicly available tumour proteomic database established from stage IV melanoma patients undergoing anti-PD-1 treatment (responder, R = 40 and non-responder, NR = 27); e-g, correlation uncleaved GSDMD peptide with NFKB1 gene targets and two sets of M1- vs. M2-like macrophage gene signatures, respectively, in stage IV melanoma patients undergoing anti-PD-1 treatment (R = 18 and NR = 6, chi-square test).

Article Snippet: To neutralize IL-1 receptor signalling, the following antibodies were used: anti-human IL-1α (1 μg/mL; clone 7D4; mabg-hil1a-3; InvivoGen), anti-human IL-1β (1 μg/mL; clone 4H5; mabg-hil1b-3; InvivoGen) and IgG1 isotype control (1 μg/mL; clone T8E5; mabg1-ctrlm; InvivoGen).

Techniques: Comparison

Journal: bioRxiv

Article Title: Dysplastic Epithelial Repair Propagates Chronic Pathology Through the Paracrine Transformation of Pulmonary Fibroblasts

doi: 10.64898/2026.04.02.716135

Figure Lengend Snippet: ( A ) Quantified levels of IL-1α from naïve fibroblast (Naïve fibro), inflammatory fibroblast (Inflam fibro), and ectopic basal cell (Basal) conditioned media as determined by ELISA. ( B ) Saa3 and Ccl2 gene expression by RT-qPCR for fibroblasts cultured on 0.5kPa plates and treated with a 5-fold titration of IL-1α added to control media, including the level determined in (A). ( C ) Graphic illustrating the strategy used to deplete conditioned media of endogenous IL-1α with antibodies, magnetic beads, and column-based separation. ( D ) Saa3 and Ccl2 gene expression by RT-qPCR for fibroblasts cultured on 0.5kPa plates in control or conditioned media depleted using IgG (CTRL IgG or CM IgG) or IL-1α nAb (CTRL nAB, CM nAb) and supplemented with additional IL-1α nAb after depletion (+Spike). [(B) and (D)] Data shows mean fold change ± SD and are pooled from 3 independent experiments (n=3 mice). P values were calculated using either (B) a ratio paired t -test or (D) a one-way ANOVA with post-hoc Turkey multiple comparison test: * P < 0.05, ** P < 0.01, **** P < 0.0001, ns: not significant.

Article Snippet: The working concentration for the following reagents can be found in the text and figure legends: Human IL-1RA Recombinant Protein (IL1Ra) (PeproTech®, 200-01RA), Mouse IL-1 alpha Recombinant Protein, (PeproTech®, 211-11A), Mouse IL-1α Neutralizing Antibody (Invivogen, Anti-mIL-1α-mIgG1 clone 6H7, mIL-1α-mab9-02), Recombinant Mouse IgG1 Isotype Control Antibody (Invivogen, Anti-β-Gal-mIgG1 clone T9C6, bgal-mab9-02), Human TGF-β 1 Recombinant Protein (PeproTech®, 100-21), Human GDF15 Recombinant Protein (PeproTech®, 120-28C), Human VEGF Recombinant Protein (PeproTech®, 450-32), Human OPN Recombinant Protein (PeproTech®, 120-35), Murine CXCL1 Recombinant Protein (PeproTech®, 250-11), Human IGFBP3 Recombinant Protein (PeproTech®, 100-08), Human TNFRSF11B Recombinant Protein (PeproTech®, 450-14), Murine CXCL5 Recombinant Protein (PeproTech®, 250-17), and Muring CCL20 Recombinant Protein (PeproTech®, 250-27).

Techniques: Enzyme-linked Immunosorbent Assay, Gene Expression, Quantitative RT-PCR, Cell Culture, Titration, Control, Magnetic Beads, Comparison

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