human ifnγ  (R&D Systems)

Journal: Molecular Therapy Oncology

Article Title: AAV immuno-gene therapy platform delivering vectorized cytokines defines a new modality for high-grade glioma treatment

doi: 10.1016/j.omton.2026.201183

Figure Lengend Snippet: Vectorized IFNβ drives durable signaling and complete tumor regression in human glioblastoma models in vivo (A) Sustained hIFNβ secretion in human GBM6 cells treated with AAV9-hIFNβ (red, MOI = 4E5 vg/cell) or recombinant hIFNβ cytokine (r-hIFNβ, purple, 47 IU/mL, equivalent to 114 pg/mL), measured by ELISA at indicated time points. 50% media washouts every 5 h for the first 20 h in the r-hIFNβ condition mimic in vivo cytokine clearance (half-life = 4–5 h). Full media exchanges were performed at 24, 48, 72, and 96 h post-treatment. (B) Number of differentially expressed genes (DEGs, p -Adj<0.01) in GBM6 cells 24–96 h post-treatment with AAV9-hIFNβ or r-hIFNβ vs. media controls. (C) Enrichment scores for type I IFN and TNFα response pathways across treatments and time points. (D) Heatmap of the top 10 IFN and TNFα response genes (Log2FC vs. media controls) in GBM6 cells treated as in (A). (E) Schematic of orthotopic PDX (SF11411) and cell line-derived xenograft ([CDX], GBM6-FLuc) studies in athymic nu/nu mice treated intratumorally with saline, AAV9-GFP, or AAV9-hIFNβ via CED. (F) Kaplan-Meier survival curves for PDX mice treated as in (E). Saline = black, AAV9-GFP (2E11 vg/brain) = blue, AAV9-hIFNβ (2E11 vg/brain) = red. Vertical dashed line = day of treatment (day 9). p < 0.04 by log-rank (Mantel-Cox) test. n = 30 (10 per treatment arm). (G) Longitudinal BLI of GBM6-FLuc tumor growth in CDX mice treated as in (E). Saline = black, AAV9-GFP (2E11 vg/brain) = blue, AAV9-hIFNβ (2E11 vg/brain) = red. Thin lines = individual mice, thick lines = geometric mean. Vertical dashed line = day of treatment (day 9). ∗ p < 0.04 by Kruskal-Wallis test with Dunn’s multiple comparisons correction on day 22. n = 30 (10 per treatment arm). (G′) Representative BLI images from each treatment group 11 days post-treatment. (H) Kaplan-Meier survival curves for CDX mice. p < 0.001 by log-rank (Mantel-Cox) test. (I) Distribution of treatment responses in CDX by BLI flux (photons/second) at day 27. Tumor free = BLI flux <2.5 × 10 5 p/s, tumor reduction = ≥30% decrease from assignment on day 9, no change = between 30% decrease and 20% increase from assignment on day 9, tumor growth = ≥20% increase from assignment on day 9, death = mice that died before day 27. (J) Dose-response analysis of AAV9-hIFNβ efficacy in CDX mice. AAV9-GFP (2E11 vg/brain) = blue, AAV9-hIFNβ hi (2E11 vg/brain) = solid red, and AAV9-hIFNβ lo (1E11 vg/brain) = dashed red. Thin lines = individual mice, thick lines = geometric mean. Vertical dashed line = day of treatment (day 9). ∗∗ p < 0.02 by Kruskal-Wallis test with Dunn’s multiple comparisons correction on day 20. n = 45 (15 per treatment arm). For data interpretation, tumor burden threshold = 2.5 × 10 5 . (J′) Representative BLI images of tumors 11 days post-treatment. (K) Kaplan-Meier survival curves from (J). p < 0.002 (AAV9-hIFNβ hi), p < 0.005 (AAV9-hIFNβ lo) by log-rank (Mantel-Cox) test compared to AAV9-GFP. (I) Distribution of treatment responses in CDX mice at day 27 by BLI flux as in (I).

Article Snippet: 24, 48, 72, and 96 h after treatment, cell supernatants were collected and IFN variant levels were measured using IFN ELISA kits following the manufacturer’s instructions (human IFNα [PBL Cat# 41135-1], human IFNβ [PBL Cat#41410], and human IFNγ [R&D Systems Cat#: DIF50C]).

Techniques: In Vivo, Recombinant, Enzyme-linked Immunosorbent Assay, Derivative Assay, Saline

Journal: Molecular Therapy Oncology

Article Title: AAV immuno-gene therapy platform delivering vectorized cytokines defines a new modality for high-grade glioma treatment

doi: 10.1016/j.omton.2026.201183

Figure Lengend Snippet: Spatial transcriptomics reveals rapid, localized transcriptional remodeling of the tumor microenvironment following vectorized hIFNβ treatment (A) Coronal brain sections from representative human GBM6-FLuc CDX mice collected pre-treatment (0 h, n = 1) or 48 h ( n = 1) after intratumoral AAV9-hIFNβ infusion (2E11 vg/brain), stained with H&E (left) and subjected to Visium Spatial Gene Expression profiling (right). Annotated clusters were assigned based on anatomical localization and marker gene expression. Dashed lines denote tumor borders. Scale bars, 1 mm. (B) Top 10 marker genes for each spatially resolved cluster identified across 0 h and 48 h datasets. Values are shown as log-normalized expression centered at 0 (Seurat “scale.data”). (C) Spatial expression of canonical human GBM tumor markers ( CD44 , VIM , TOP2A , and NOTCH1 ) delineating tumor and peri-tumor regions before (0 h) (top) and after (bottom) (48 h) AAV9-hIFNβ treatment. (D) Expression maps of the human IFNβ payload and hallmark IFN-response genes ( CXCL10 , IFIT1 , and IFIT2 ), demonstrating tumor-restricted transgene expression and induction of an IFN-specific transcriptional program within 48 h. (E) Spatial expression of host mouse immune-response genes ( Gfap , Ifitm3 , and Irf7 ) showing localized activation of astroglial and innate immune pathways proximal to the tumor. (F) Integrated datasets (0 and 48 h) visualized using canonical correlation analysis (CCA), showing distinct clustering of tumor and peri-tumor regions (left) and enrichment of IFN-response gene module expression (right). (G) Volcano plot depicting differential gene expression between 0- and 48-h tumor clusters. Red, IFN-response genes; gray, other significantly upregulated genes ( p -Adj <0.01); blue, non-significant. (H) Top enriched Gene Ontology (GO) terms among upregulated genes in 48-h tumor cells, highlighting interferon and inflammatory response pathways (∗∗ p -Adj <0.01; ∗ p -Adj <0.05).

Article Snippet: 24, 48, 72, and 96 h after treatment, cell supernatants were collected and IFN variant levels were measured using IFN ELISA kits following the manufacturer’s instructions (human IFNα [PBL Cat# 41135-1], human IFNβ [PBL Cat#41410], and human IFNγ [R&D Systems Cat#: DIF50C]).

Techniques: Spatial Transcriptomics, Staining, Gene Expression, Marker, Expressing, Activation Assay

Journal: Redox Report : Communications in Free Radical Research

Article Title: Bilirubin reduces mortality in sepsis models by inhibiting NOX2-mediated formation of neutrophil extracellular traps

doi: 10.1080/13510002.2026.2664962

Figure Lengend Snippet: Effects of bilirubin against CLP-induced sepsis in mice. (A) Mice were subjected to CLP surgery and intravenously injected with bilirubin (40 mg/kg) or vehicle. Mice were monitored at 12-h intervals to check the survival rates. (B) After CLP surgery, all mice received intraperitoneal injection of an antibiotics mixture (ceftriaxone 50 mg/kg and metronidazole 35 mg/kg). Mice were then intravenously administered either bilirubin (40 mg/kg) or vehicle. The state of survival was monitored for 96 h. The survival rates between the bilirubin and the vehicle groups were statistically analyzed using the Mantel-Cox test ( N = 15). (C, D) Biomarkers for NETosis were measured in the plasma of sham or CLP mice treated with vehicle or bilirubin 12 h after injection. Cell-free DNAs were stained with PicoGreen (C) and the H3cit levels were assessed using the ELISA kit (D). (E, F) Pro-inflammatory cytokines TNF-α (E) and IFN-γ (F) were analyzed in the plasma of the mice using specific ELISA kits. Each bar represents the mean ± SEM ( N = 5–8). Asterisks and numbers represent statistical significance and p- values by Mann–Whitney U test, respectively.

Article Snippet: Biochemical analyses were performed using the Human IFN-γ Quantikine ELISA Kit (R&D Systems; Minneapolis, MN), the Human TNF-α Quantikine ELISA Kit (R&D Systems; Minneapolis, MN), the Citrullinated Histone H3 ELISA Kit (Cayman Chemical; Ann Arbor, MI), and the Quant-iTTM PicoGreenTM dsDNA Assay Kit (Invitrogen; Waltham, MA).

Techniques: Injection, Clinical Proteomics, Staining, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

Journal: Redox Report : Communications in Free Radical Research

Article Title: Bilirubin reduces mortality in sepsis models by inhibiting NOX2-mediated formation of neutrophil extracellular traps

doi: 10.1080/13510002.2026.2664962

Figure Lengend Snippet: Effects of bilirubin against LPS-induced sepsis in mice. (A) Mice were intraperitoneally injected with 40 mg/kg of LPS and intravenously injected with bilirubin (40 mg/kg) or vehicle. Mice were monitored at 12-h interval to check the survival rates. The survival rates were statistically analyzed using the Mantel-Cox test ( N = 15). (B, C) NETosis markers were measured in the plasma of PBS control or LPS-treated mice 12 h after bilirubin or vehicle injection. The plasma levels of cell-free DNAs and H3cit were analyzed using PicoGreen (B) and the ELISA kit (C), respectively. (D, E) Pro-inflammatory cytokines TNF-α (D) and IFN-γ (E) were analyzed in the plasma using specific ELISA kits. Each bar represents the mean ± SEM ( N = 5–8). Asterisks and numbers represent statistical significance and p- values by Mann–Whitney U test, respectively.

Article Snippet: Biochemical analyses were performed using the Human IFN-γ Quantikine ELISA Kit (R&D Systems; Minneapolis, MN), the Human TNF-α Quantikine ELISA Kit (R&D Systems; Minneapolis, MN), the Citrullinated Histone H3 ELISA Kit (Cayman Chemical; Ann Arbor, MI), and the Quant-iTTM PicoGreenTM dsDNA Assay Kit (Invitrogen; Waltham, MA).

Techniques: Injection, Clinical Proteomics, Control, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

Journal: iScience

Article Title: Single-cell transcriptomic analysis reveals intra-tumoral heterogeneity and immunotherapy strategies in high-grade serous ovarian cancer

doi: 10.1016/j.isci.2026.115266

Figure Lengend Snippet: Validation of tumor-associated marker genes (A) Expression of tumor-associated markers in epithelial cells from different cancers. (B) Scoring for major cell types based on tumor-associated markers. (C) Scoring of tumor diagnostic marker-related genes for major cell types. (D) Scoring of EV marker genes for major cell types. (E) Expression levels of co-stimulatory molecules and MHC molecules in different epithelial cell clusters. (F) Expression levels of top 2 genes in different epithelial cell clusters. (G and H) mIHC images displaying the expression of IFIT1 , epithelial marker (pan-CK), MHC class Ⅰ marker (B2M), and CD8 + T cell markers (CD3 + /CD8 + ) in HGSOC patients with (G) unfavorable prognosis and (H) favorable prognosis; n = 50. (I–K) CD8 + T cells (effectors) were incubated with IFIT1 + or IFIT1 − tumor cells (targets) at a 10:1 effector-to-target (E:T) ratio for 72 h. (I) Schematic of the in vitro culture model. (J) Representative flow cytometry plots of the results of the apoptosis assays and the quantification of apoptosis assays based on their Annexin V + percentages. (K) Levels of IFN-γ and granzyme B in the supernatant, measured via ELISA. Data are presented as the mean ± SEM, n = 5 pairs, including 5 patients and 5 healthy volunteers (J and K). ∗ p < 0.05, ∗∗ p < 0.01, two-tailed Student’s t test.

Article Snippet: The conditioned medium (CM) of co-cultured CD8 + T cells and CD133 + IFIT1 + or CD133 + IFIT1 - cells were analyzed to measure IFN-γ or GZMB levels using ELISA kits (R&D Systems, QK285, DY2906-05).

Techniques: Biomarker Discovery, Marker, Expressing, Diagnostic Assay, Incubation, In Vitro, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: Scientific Reports

Article Title: Diagnostic and immunological roles of leptin gene rs7799039 polymorphism and cytokines in COVID-19, HCV, and dual infection

doi: 10.1038/s41598-026-35418-4

Figure Lengend Snippet: Flowchart of participant recruitment and study design. A total of 228 participants were enrolled and categorized equally into four groups: HCV-COVID-19 co-infected ( n = 57), COVID-19 only ( n = 57), HCV only ( n = 57), and healthy controls ( n = 57). Each participant provided a nasopharyngeal swab and a blood sample for molecular (RT-PCR), serological, cytokine (ELISA), and genetic (TaqMan SNP genotyping) analyses.

Article Snippet: IFN-γ levels were determined using the R&D Systems Quantikine ® ELISA kit (Catalog No. DIF50), which provides a detection range of 15.6–1000 pg/mL, a sensitivity of < 8 pg/mL, intra-assay CVs of 2.6–4.7%, and inter-assay CVs of 3.7–7.8%.

Techniques: Infection, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

Journal: The Journal of Clinical Investigation

Article Title: Bispecific antibodies and CAR T cells targeting a TP53 mutation–associated neoantigen show discordant affinity requirements

doi: 10.1172/JCI192885

Figure Lengend Snippet: ( A and B ) Variants were expressed as scDbs. 5 × 10 4 primary human T cells were cocultured overnight with 2.5 × 10 4 T2A3 target cells pulsed with ( A ) 10 nM or ( B ) varying concentrations of the p53 R175H or R175WT 9mer peptide in the presence of 1 nM scDb. ( A ) IFN-γ production and ( B ) percent cytotoxicity compared with no scDb control by CellTiterGlo assay ( n = 3 biological replicates, single experiment). ( C and D ) 5 × 10 4 T cells were cocultured with 2.5 × 10 4 luciferase + KMS26 target cells (p53 R175H or p53 KO) and scDb for 18 hours. ( C ) IFN-γ production. ( D ) Percent cytotoxicity measured by SteadyGlo assay ( n = 3 biological replicates, single experiment). All data are shown as the mean ± SD. ( E and F ) Top variants were combined into double-mutant scDbs. ( E ) scDb was applied to plates coated with CD3ε/δ heterodimer, or R175H/A2 or R175WT/A2 monomer. Relative binding was calculated by normalizing mean monomer binding to mean CD3 binding ( n = 3 technical replicates, single experiment). ( F ) 3 × 10 4 T cells were cocultured with 1.5 × 10 4 KMS26 target cells in the presence of scDb for 19 hours. Percent cytotoxicity by SteadyGlo assay ( n = 3 biological replicates per condition, single experiment).

Article Snippet: IFN-γ production was quantified with the Quantikine Human IFN-γ ELISA Kit (DIF50C, R&D Systems).

Techniques: Control, Luciferase, Mutagenesis, Binding Assay

Journal: The Journal of Clinical Investigation

Article Title: Bispecific antibodies and CAR T cells targeting a TP53 mutation–associated neoantigen show discordant affinity requirements

doi: 10.1172/JCI192885

Figure Lengend Snippet: ( A ) Single-cycle binding kinetics for each H2 variant scDb binding to the p53 R175H/HLA-A*02:01 monomer measured by SPR. Dashed line indicates curve fit. ( B ) Coculture design testing scDb cytotoxicity against isogenic p53 R175H and p53 –/– cell lines or p53 R175WT cell lines. ( C and D ) 5 × 10 4 T cells were cocultured with 2.5 × 10 4 KMS26 target cells in the presence of 0–3 nM scDb for 20 hours. ( C ) Percent cytotoxicity compared with no scDb was measured by SteadyGlo. ( D ) IFN-γ production ( n = 3, representative of 3 independent experiments, curve fitting and EC 50 s determined by 5-parameter log fit). ( E ) 1 × 10 4 GFP + TYK-nu target cells were incubated with 2 × 10 4 T cells and 0.01 nM scDb and counted by live-cell imaging ( n = 3, representative of 2 independent experiments). ( F ) Mean GFP + target cells at 120 hours (ordinary 1-way ANOVA with Tukey’s multiple comparisons). ( G and H ) 5 × 10 4 T cells were cocultured with 2.5 × 10 4 target cells and 0.01 nM scDb for 20 hours. Percent cytotoxicity compared with no scDb was measured by SteadyGlo ( n = 3, representative of 3 independent experiments, 1-way ANOVA with Holm-Šidák multiple comparisons). All data are shown as the mean ± SD.

Article Snippet: IFN-γ production was quantified with the Quantikine Human IFN-γ ELISA Kit (DIF50C, R&D Systems).

Techniques: Binding Assay, Variant Assay, Incubation, Live Cell Imaging

Journal: The Journal of Clinical Investigation

Article Title: Bispecific antibodies and CAR T cells targeting a TP53 mutation–associated neoantigen show discordant affinity requirements

doi: 10.1172/JCI192885

Figure Lengend Snippet: ( A ) Illustration of the patient-derived TCR AV6/BV11 compared with the H2 STAR, where the H2 variable heavy (VH) and variable light (VL) chains are linked to the TCRα constant domain (TRAC) and TCRβ constant domain (TRBC), respectively. ( B ) Transgenic T cells were generated by CRISPR knock-out at TRBC1/TRBC2 and knockin at TRAC using dsDNA homology-directed repair templates (HDRTs). HA, homology arm. ( C ) T cells expressing STARs were characterized by flow cytometry. Representative histograms of p53 R175H/HLA-A2 tetramer bound to edited (CD3 + tetramer) CD4 + and CD8 + T cells are shown. ( D ) Tetramer staining on CD4 + and CD8 + T cells was compared across n = 5 independent experiments by normalizing tetramer MFI to the tetramer MFI for CD4 + AV6/BV11 TCR T cells for each experiment (2-way ANOVA with Dunnett’s multiple comparisons to H2 29.5 ). ( E – H ) T cell activation at varying antigen densities was tested in a peptide-pulsing coculture. 8 × 10 3 edited T cells were cocultured with 4 × 10 4 peptide-pulsed T2 cells for 24 hours. ( E ) T cell–mediated cytotoxicity was measured by CellTiterGlo viability assay. ( F ) IFN-γ production. ( G ) Cytotoxicity and ( H ) IFN-γ were compared at selected concentrations from E and F (1-way ANOVA with Dunnett’s multiple comparisons, n = 3 biological replicates, representative of 3 independent experiments). All data are shown as the mean ± SD.

Article Snippet: IFN-γ production was quantified with the Quantikine Human IFN-γ ELISA Kit (DIF50C, R&D Systems).

Techniques: Derivative Assay, Transgenic Assay, Generated, CRISPR, Knock-Out, Knock-In, Expressing, Flow Cytometry, Staining, Activation Assay, Viability Assay

Journal: The Journal of Clinical Investigation

Article Title: Bispecific antibodies and CAR T cells targeting a TP53 mutation–associated neoantigen show discordant affinity requirements

doi: 10.1172/JCI192885

Figure Lengend Snippet: ( A ) 5 × 10 4 edited T cells were cocultured with 1 × 10 5 GFP-expressing NALM6 R175H or NALM6 WT target cells for 2 days. ( B and C ) 1.2 × 10 4 edited T cells were cocultured with 2.4 × 10 4 KMS26 target cells for 24 hours. ( B ) Percent cytotoxicity by SteadyGlo assay compared with the TRAC/TRBC-KO condition. ( C ) IFN-γ production ( n = 3 per condition, 2 independent experiments, 2-way ANOVA with Dunnett’s multiple comparisons). ( D – F ) STAR T cell multiple stimulation assay. 4 × 10 3 edited T cells were cocultured with 1.6 × 10 4 KMS26 target cells in replicate plates and rechallenged with 3.2 × 10 4 target cells or assayed by flow cytometry on days 2, 4, and 6. ( D ) Live GFP + target cells; ( E ) CellTrace Violet (CTV) dilution, indicative of cell division, in NGFR + -edited T cells relative to day 2; and ( F ) counts of NGFR + T cells ( n = 3 per condition, 3 independent experiments). ( G – I ) T cell killing of KLE target cells was observed by live-cell imaging (aggregate of 3 independent experiments per T cell type, Brown-Forsythe and Welch’s ANOVA with Dunnett’s T3 multiple comparisons to H2 3.3 ). ( G ) Time from T cell interaction to target cell blebbing (arrows indicate T cells). ( H ) Time from target cell blebbing to T cell detachment. ( I ) Total duration of interaction. ( J – L ) 5 × 10 4 edited T cells were cocultured with 1 × 10 5 NALM6 R175H or NALM6 WT target cells for 17 hours and assayed by flow cytometry ( n = 3 per condition, 2 independent experiments). ( J ) Percent CD69 + CD137 + of live T cells (2-way ANOVA with Dunnett’s multiple comparisons to H2 29.5 ). ( K ) Percent IFN-γ + of live T cells (2-way ANOVA with Dunnett’s multiple comparisons to H2 29.5 ). ( L ) MFI of IFN-γ in CD8 + T cells (ordinary 1-way ANOVA with Šidák’s multiple-comparison test to H2 29.5 ). All data are shown as the mean ± SD.

Article Snippet: IFN-γ production was quantified with the Quantikine Human IFN-γ ELISA Kit (DIF50C, R&D Systems).

Techniques: Expressing, Flow Cytometry, Live Cell Imaging, Comparison