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