Journal: Cells

Article Title: Bone Marrow-Derived IL-1Ra Increases TNF Levels Poststroke

doi: 10.3390/cells10040956

Figure Lengend Snippet: Effect of bone marrow (BM) cells on ischemia-induced proteins in brain and serum. ( A ) BM cell treatment 30 min after tMCAo reduces infarct volumes (IFV) in LM–LM and Tg–LM mice as well as brain IL-1β levels (pg/mg) in Tg–LM mice 24 h after tMCAo, compared to LM tMCAo mice (Table with extracted data from ) (the experimental overview was created with BioRender.com). ( B – K ) Comparison of TNF ( C ), IL-4 ( D ), IL-10 ( E ), CXCL1 ( F ), IL-12p70 ( G ), IL-5 ( H ), IL-6 ( I ), IFNγ ( J ) and IL-2 ( K ), levels in the brain (pg/mg) and serum (pg/mL) of unlesioned controls and in LM, LM–LM, and Tg–LM mice 24 h after tMCAo. Data are presented as mean ± SEM ( n = 10–12/group). One-way ANOVA, followed by Sidak’s post hoc tests, was performed. Significance indicated as * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. N.D, not detected.

Article Snippet: Electrochemiluminescence analysis was performed on brain, serum, cells, and media using the MSD Mouse Proinflammatory V-Plex Plus Kit (IFNγ, IL-2, IL-4, IL-5, IL-6, IL-10, IL-12p70, CXCL1, TNF; K15012C, Mesoscale Discovery), the U-Plex Kit (TNFR1; K15069L, Mesoscale), and the R-Plex kit (TNFR2; K150ZSR-2, Mesoscale Discovery) according to the manufacturer’s instructions.

Techniques: Comparison

Journal: Molecular therapy : the journal of the American Society of Gene Therapy

Article Title: Exploiting viral infection/vaccination to focus high-affinity T cell populations into tumors using oncolytic viro-immunotherapy

doi: 10.1016/j.ymthe.2025.08.023

Figure Lengend Snippet: (A) C57BL6 mice were intramuscularly (IM) vaccinated 30 days prior to B16-ACE2 tumor inoculation with either VSV (10 8 PFU) or Ad (10 10 vp). Tumors were treated IT 8, 10, and 12 days after tumor seeding, with VSV expressing SARS-Mem or emGFP (10 8 PFU/injection). (B) Survival with time is shown; ** p ≤ 0.01, *** p ≤ 0.001; log rank test with Grehan-Breslow-Wilcoxon test. (C) Tumor volumes across different treatment groups. The majority of mice treated with VSV-Mem: VSV-Mem reached endpoint by day 40. The only mouse surviving developed a small tumor 26 days post-tumor cell challenge, which completely regressed by day 38. (D and E) IFN-γ ELISpot from CD8 + cells isolated from splenocytes of different treatment groups re-stimulated in vitro with live B16 melanoma (D) or irrelevant CT2A glioma (E) cells as targets (E:T ratio 10:1). Error bars denote SD. ** p ≤ 0.01, *** p ≤ 0.001; **** p ≤ 0.0001; ns, statistically non-significant; one-way ANOVA.

Article Snippet: For ELISpot assays (Mouse Interferon-γ ELISpot Plus , MABTECH AB, Nacka Strand, Sweden), target tumor cells (B16, B16-EGFRvIII, CT2A-EGFRvIII, or irrelevant CT2A glioma) were treated with 50 U recombinant (r)IFN-γ for 24 h prior to quantification of surface-expressed K2b/SIINFEKL or co-culture with T cells to increase the frequency of cells expressing major histocompatibility complex class I (murine rIFN-γ; catalog no. 14-8311-63, eBioscience, San Diego, CA).

Techniques: Expressing, Injection, Enzyme-linked Immunospot, Isolation, In Vitro

Journal: Molecular therapy : the journal of the American Society of Gene Therapy

Article Title: Exploiting viral infection/vaccination to focus high-affinity T cell populations into tumors using oncolytic viro-immunotherapy

doi: 10.1016/j.ymthe.2025.08.023

Figure Lengend Snippet: (A) ACE2-expressing tumor cells infected with VSV-S generate large multi-nucleated syncytia (one example highlighted with red arrows) 48 h post-infection. (B) C57BL6 mice were vaccinated IM 30 days prior to seeding of B16-ACE2 tumors with SC-Ad (10 10 vp) expressing either Mem or S. At 8, 10, 12, 17, 19, and 21 days later tumors were treated IT with either VSV (10 8 PFU) or SC-Ad (10 10 vp) expressing Mem or S. (C) Survival with time is shown; *** p ≤ 0.001, **** p ≤ 0.0001; ns, statistically non-significant; log rank test with Grehan-Breslow-Wilcoxon test. (D) Tumor volumes across different treatment groups. (E) IFN-γ ELISpot from CD8 + cells isolated from splenocytes of different treatment groups re-stimulated in vitro with live B16 melanoma (E) or irrelevant CT2A glioma (F) cells as targets (E:T ratio 10:1). Error bars denote SD. **** p ≤ 0.0001; ns, statistically non-significant; one-way ANOVA.

Article Snippet: For ELISpot assays (Mouse Interferon-γ ELISpot Plus , MABTECH AB, Nacka Strand, Sweden), target tumor cells (B16, B16-EGFRvIII, CT2A-EGFRvIII, or irrelevant CT2A glioma) were treated with 50 U recombinant (r)IFN-γ for 24 h prior to quantification of surface-expressed K2b/SIINFEKL or co-culture with T cells to increase the frequency of cells expressing major histocompatibility complex class I (murine rIFN-γ; catalog no. 14-8311-63, eBioscience, San Diego, CA).

Techniques: Expressing, Infection, Enzyme-linked Immunospot, Isolation, In Vitro

Journal: Molecular therapy : the journal of the American Society of Gene Therapy

Article Title: Exploiting viral infection/vaccination to focus high-affinity T cell populations into tumors using oncolytic viro-immunotherapy

doi: 10.1016/j.ymthe.2025.08.023

Figure Lengend Snippet: (A) B16-EGFRvIII tumors were seeded in C57Bl/6 k18-hACE2 mice (n = 7–8/group). On days 7, 9, and 11 mice were either left unimmunized or were immunized with an S-derived peptide library (1 μg/mouse/injection). On days 14, 16, and 18, groups were treated with unloaded anti-EGFRvIII CAR-T cells (10 7 CAR-T cells/injection), with CAR-T cells loaded ex vivo with SC-Ad-GFP or with CAR-T cells loaded ex vivo with SC-Ad-S (10 7 CAR-T cells, loaded ex vivo at 4°C, MOI 1.0). (B) Survival with time is shown; *** p ≤ 0.001; **** p ≤ 0.0001; log rank test with Grehan-Breslow-Wilcoxon test. (C and D) IFN-γ ELISpot from splenocytes isolated from different treatment groups re-stimulated in vitro with live B16 melanoma (C) or CAR target antigen expressing CT2A-EGFRvIII glioma (D) cells as targets (E:T ratio 1:1). Error bars denote SD. *** p ≤ 0.001; **** p ≤ 0.0001; ns, statistically non-significant; one-way ANOVA. (E and F) IFN-γ ELISpot from splenocytes recovered from different treatment groups re-stimulated in vitro with S-specific peptide library or irrelevant ovalbumin-derived SIINFEKL peptide; Error bars denote SD. * p ≤ 0.05, ** p ≤ 0.001, **** p ≤ 0.0001; ns, statistically non-significant; one-way ANOVA.

Article Snippet: For ELISpot assays (Mouse Interferon-γ ELISpot Plus , MABTECH AB, Nacka Strand, Sweden), target tumor cells (B16, B16-EGFRvIII, CT2A-EGFRvIII, or irrelevant CT2A glioma) were treated with 50 U recombinant (r)IFN-γ for 24 h prior to quantification of surface-expressed K2b/SIINFEKL or co-culture with T cells to increase the frequency of cells expressing major histocompatibility complex class I (murine rIFN-γ; catalog no. 14-8311-63, eBioscience, San Diego, CA).

Techniques: Derivative Assay, Injection, Ex Vivo, Enzyme-linked Immunospot, Isolation, In Vitro, Expressing

Journal: Cell Reports Medicine

Article Title: Coupling IL-2 with IL-10 to mitigate toxicity and enhance antitumor immunity

doi: 10.1016/j.xcrm.2025.102257

Figure Lengend Snippet: Coupling and targeting IL-2 and IL-10 together retains broad immune activation and increases anti-tumor potency in mice (A–M) C57BL/6 mice (C57BL/6 and IFNγ-knockout mice in H) were implanted subcutaneously with a human EGFR-expressing B16F10 cell line (B16F10 hEGFR+ ). B16F10 hEGFR+ tumor-bearing mice were treated three times per week with vehicle control or the reagent(s) indicated. (A–F, H–L) Data are representative of the tumor volumes (mm 3 ) from 4 to 15 animals per group. (A) Mice treated with vehicle, IL-2 (0.4 mg/kg), IL-10 (1.0 mg/kg), or both ( n = 10). Statistical analyses were performed by Mann-Whitney test. (B) Mice treated with vehicle, IL-2 (scFv) (0.4 mg/kg), IL-10 (scFv) (1.0 mg/kg), or both ( n = 10). Statistical analyses were performed by two-way ANOVA with Tukey’s multiple comparisons test. (C) Mice treated with vehicle, IL-2 (EGFR) (0.4 mg/kg), IL-10 (EGFR) (1.0 mg/kg), or both ( n = 10). Statistical analyses were performed by two-way ANOVA with Tukey’s multiple comparisons test. (D) Mice treated with vehicle control or varying doses (0–6 mg/kg) of DK2 10 (EGFR) ( n = 7). Statistical analyses were performed by two-way ANOVA with Tukey’s multiple comparisons test. (E) Mice treated with vehicle, DK2 10 (scFv) (2.0 mg/kg), or DK2 10 (EGFR) (2.0 mg/kg) ( n = 9–10). Statistical analyses were performed by Mann-Whitney test. (F) Mice treated with vehicle, DK2 10 (EGFR) (2.0 mg/kg), or varying doses of IL-2 (0–4.0 mg/kg) ( n = 4–7). Statistical analyses were performed by two-way ANOVA with Tukey’s multiple comparisons test. (G) Mice treated with vehicle, DK2 10 (EGFR) (2.0 mg/kg), or IL-2 (2.0 mg/kg) ( n = 4–7). Serum was collected, and cytokine concentrations are reported. Statistical analyses were performed by Student’s t test. Results are reported as the mean ± SD. (H) Wild-type or IFNγ-knockout mice were treated with vehicle or DK2 10 (EGFR) (2.0 mg/kg) ( n = 7–9). Statistical analyses were performed by Welch’s t test. (I–K) Mice treated with vehicle or DK2 10 (EGFR) (2.0 mg/kg) with and without cell ablation of CD8 + (I), CD4 + (J), or NK cells (K) ( n = 5–6). Statistical analyses were performed by Mann-Whitney test. (L) Mice treated with vehicle or DK2 10 (EGFR) (2.0 mg/kg) with and without FTY720 ( n = 11–15). Statistical analyses were performed by Mann-Whitney test. (M–P) Mice were treated with vehicle or DK2 10 (EGFR) (2.0 mg/kg) for 6–8 days ( n = 4). (M) The frequencies of CD4 + and CD8 + T cells among TILs. (N) Following treatment, CD8 + Tem (CD3 + /CD8 + /CD44 + /CD62L − ) TILs were sorted, and gene expression was performed by nCounter analysis. Genes with a fold change of 1.5 from control animals and p value ≤0.05 are reported. (O) The oxygen consumption rate of isolated CD8 + TILs ( n = 9). (P) Isolated CD8 + TILs from treated mice were rested overnight before co-culturing with B16F10 hEGFR+ cells, and an Elispot was performed to detect IFNγ-producing tumor-reactive cells ( n = 5–9). Statistical analyses were performed by Welch’s t test. (Q) Tumor volumes (mm 3 ) from mice bearing LL2 hEGFR+ tumors treated with vehicle or DK2 10 (EGFR) with or without anti-PD-1 (2.5 mg/dose) for 1 week ( n = 9). Statistical analyses were performed by Welch’s t test. (R) A patient-derived xenograft (PDX) model (non-small cell lung cancer) was established by subcutaneous implant of patient tumor cells, and mice were treated with vehicle, anti-PD-1 (10 mg/kg; every 5 days intraperitoneally), or DK2 10 (EGFR) (0.5 or 2.0 mg/kg, three times a week subcutaneously) ( n = 2). Tumor volumes are reported (mm 3 ). Statistical analyses were performed by Welch’s t test. (ns, not significant; p value; ∗ p ≤ 0.05; ∗∗ p ≤ 0.01; ∗∗∗ p ≤ 0.001; ∗∗∗∗ p ≤ 0.0001). Results are reported as the mean ± SEM, unless otherwise noted.

Article Snippet: ELISpot Plus: Mouse IFN-γ (HRP) , Mabtech , Cat# 3321-4HPT-2.

Techniques: Activation Assay, Knock-Out, Expressing, Control, MANN-WHITNEY, Gene Expression, Isolation, Enzyme-linked Immunospot, Derivative Assay