Journal: Molecular Therapy Advances

Article Title: Efficacy and safety of SENS-501, a dual-AAV otoferlin gene therapy, for DFNB9 congenital deafness

doi: 10.1016/j.omta.2026.201762

Figure Lengend Snippet: Intracochlear injection of SENS-501 in NHPs results in a mild humoral and an undetectable cellular response to the capsid (A) AAV8 anti-drug antibody (ADA) titers measured in serum at pre-dose and at 16, 29, and 92 days post-vector administration in NHPs. (B) Anti-AAV8 neutralizing antibodies (NAb) titers measured in serum at pre-dose and at 16, 29, and 92 days post-vector administration in NHPs. (C and D) IFN-γ spot forming units (SFUs) measured by ELISpot assay at 29 (C; left) and 92 (D; right) days post-injection. Peripheral blood mononuclear cells (PBMCs) from the indicated groups were stimulated with three different AAV8 peptide pools and a positive control (PMA/ionomycin). The dotted line represents the assay-specific positivity threshold. Each dot represents one animal. Bars represent the mean ± SEM.

Article Snippet: After the incubation, detection was performed with a monoclonal anti-monkey IFN-γ antibody (Monkey IFN-γ ELISpot Pro Kit, Mabtech) coupled with alkaline phosphatase and incubated with BCIP/NBT (5-bromo-4-chloro-3-indolyl-1-phosphate / nitroblue tetrazolium) substrate to detect secreted IFN-γ.

Techniques: Injection, Plasmid Preparation, Enzyme-linked Immunospot, Positive Control

Journal: Poultry Science

Article Title: A novel self-amplified RNA vaccine co-expressing NA and HA1 delivered by Salmonella confers potent protection against H9N2 influenza in chickens

doi: 10.1016/j.psj.2026.107072

Figure Lengend Snippet: Intracellular cytokine production. Chicken splenic lymphocytes were isolated for analysis. Cell proliferation was assessed using CCK-8 analysis with ConA (A), mixed HA1 peptides (B), and NA protein (C). Additionally, the production of IFN-γ by splenic T lymphocytes was measured via an ELISpot assay, utilizing NA and HA1 proteins as stimulators for 36 h (D).

Article Snippet: IFN-γ production was assessed using a commercial Chicken IFN-γ ELISpot kit (Mabtech, Sweden).

Techniques: Isolation, CCK-8 Assay, Enzyme-linked Immunospot

Journal: Poultry Science

Article Title: A novel self-amplified RNA vaccine co-expressing NA and HA1 delivered by Salmonella confers potent protection against H9N2 influenza in chickens

doi: 10.1016/j.psj.2026.107072

Figure Lengend Snippet: Intracellular cytokine production. The intracellular mRNA expression levels of IL-4 (B, D) and IFN-γ (A, C)—as well as the relative concentrations of these cytokines in cell culture supernatants stimulated by the NA peptide (E, F) or HA1 protein (G, H) for 48 h—were determined using qRT-PCR and ELISA, respectively. Data are expressed as the mean ± SEM and analyzed using one-way ANOVA (* P < 0.05, ** P < 0.01, and *** P < 0.001; n = 4).

Article Snippet: IFN-γ production was assessed using a commercial Chicken IFN-γ ELISpot kit (Mabtech, Sweden).

Techniques: Expressing, Cell Culture, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: Molecular Therapy Oncology

Article Title: Induced pluripotent stem cell-derived dendritic cells potentiate the efficacy of interleukin-12-expressing oncolytic HSV-1

doi: 10.1016/j.omton.2026.201252

Figure Lengend Snippet: Immunological assessments of the combination of T-mfIL12 and immature BMDCs (A) Immunohistochemical analysis of representative MB49 tumors from the indicated treatment groups. Increased intratumoral infiltration of CD8 + lymphocytes was most prominent in the T-mfIL12+BMDC group, followed by the T-01+BMDC group. In both groups, elevated levels of CD4 + lymphocyte infiltration were also observed in the same tumor sections. Other groups, except for the mock group, showed moderate levels of CD4 + infiltration. Scale bars: 100 μm. (B) ELISpot assay for IFN-γ expression revealed the highest number of IFN-γ-producing splenocytes in the T-mfIL12+BMDC group, followed by the T-01+BMDC group. Monotherapies with BMDC, T-01, or T-mfIL12, as well as the mock group, did not result in significant increases. n = 3 per group; all samples were analyzed in triplicate. Error bars represent standard deviation (SD). ∗ p < 0.05, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: Cells were plated in triplicates at a density of 1 × 10 5 cells per well in 50 μL of RPMI-1640 medium supplemented with 10% FCS, 2% penicillin-streptomycin, and 50 μM 2-ME, using a MultiScreen 96-well plate (Millipore, USA) pre-coated with anti-mouse IFN-γ monoclonal antibodies from the Mouse IFN-γ ELISpot Kit (AN18; Mabtech AB, cat. #3321-2A).

Techniques: Immunohistochemical staining, Enzyme-linked Immunospot, Expressing, Standard Deviation

Journal: Molecular Therapy Oncology

Article Title: Induced pluripotent stem cell-derived dendritic cells potentiate the efficacy of interleukin-12-expressing oncolytic HSV-1

doi: 10.1016/j.omton.2026.201252

Figure Lengend Snippet: Immunological assessments of the combination of T-mfIL12 and immature iPSDCs (A) Immunohistochemical analysis of representative MB49 tumors from the indicated treatment groups. Increased intratumoral infiltration of CD8 + lymphocytes was observed in both the T-mfIL12+BMDC and T-mfIL12+iPSDC groups. In both groups, CD4 + lymphocyte infiltration was also more prominently observed in the same tumor sections. Other treatment groups, excluding the mock group, exhibited moderate levels of CD4 + T cell infiltration. Scale bars: 100 μm. (B) ELISpot assay showed a high and comparable increase in IFN-γ-producing splenocytes in both the T-mfIL12+BMDC and T-mfIL12+iPSDC groups. n = 3 mice per group; all samples were analyzed in triplicate. Error bars represent standard deviation (SD). ∗∗∗∗ p < 0.0001; NS, not significant.

Article Snippet: Cells were plated in triplicates at a density of 1 × 10 5 cells per well in 50 μL of RPMI-1640 medium supplemented with 10% FCS, 2% penicillin-streptomycin, and 50 μM 2-ME, using a MultiScreen 96-well plate (Millipore, USA) pre-coated with anti-mouse IFN-γ monoclonal antibodies from the Mouse IFN-γ ELISpot Kit (AN18; Mabtech AB, cat. #3321-2A).

Techniques: Immunohistochemical staining, Enzyme-linked Immunospot, Standard Deviation

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