mouse ifn γ elispot kit  (Mabtech Inc)

Journal: Smart Molecules

Article Title: Endocytosis‐independent cytosolic entry of messenger RNA via fluorous bilayer zippering attenuating Toll‐like receptor signaling and enables ischemic tissue salvage

doi: 10.1002/smo2.70085

Figure Lengend Snippet: Biocompatibilities and immunogenicity of mRNA‐encapsulating polyplexes. (a) Cell viabilities of HUVECs upon 24 h incubation in presence of a variety of mRNA‐encapsulating polyplexes (mean ± s.d., n = 4, ** p < 0.01, student t test). (b) Hemolytic activities of sheep red blood cells upon 2 h incubation with varied concentrated delivery materials (*** p < 0.005; Student's t ‐test, mean ± s.d., n = 4). (c) confocal laser scanning microscopy (CLSM) measurement for assessment of overall cellular internalization of mRNA‐encapsulating polyplexes into RAW264.7 cells. (d) Interferon‐β (IFN‐β): at 4 h post incubation, the expression levels of inflammatory molecules were measured with qRT‐PCR. (e) Interleukin‐8 (IL‐8): at 4 h post incubation, the expression levels of inflammatory molecules were measured with qRT‐PCR. (** p < 0.01, *** p < 0.005; Student's t ‐test, n = 4).

Article Snippet: Culture supernatants (100 μL/well) were harvested and assayed for IFN‐α using the Mouse IFN‐α ELISA Kit (PBL Interferon Source) according to the manufacturer's instructions.

Techniques: Immunopeptidomics, Incubation, Confocal Laser Scanning Microscopy, Expressing, Quantitative RT-PCR

Journal: Smart Molecules

Article Title: Endocytosis‐independent cytosolic entry of messenger RNA via fluorous bilayer zippering attenuating Toll‐like receptor signaling and enables ischemic tissue salvage

doi: 10.1002/smo2.70085

Figure Lengend Snippet: Revascularization in hindlimbs by local dosage of mVEGF‐encapsulating polyplexes. (a) Therapeutic scheme. (b) Anatomy of the established hindlimb ischemia model. Ligations were made in the femoral artery at the proximal and distal sites. (c) Angiogenesis in mouse hindlimbs post ligation. (d) Visualization of blood flow by Laser Speckle Flowgraphy on Day 28 post‐dosage of mVEGF therapeutics (mVEGF: 10 μg). The magnified inset images captured by intravital confocal laser scanning microscopy (CLSM), revealing vasculature details by intravenous dosage of FITC‐dextran (MW: 10 kDa). (e) Estimation of blood perfusion volume based on quantification by laser speckle flowgraphy on Day 28 post‐dosage of mVEGF therapeutics (mVEGF: 10 μg). The data were represented as the mean ± standard deviations (s.d.) ( n = 5). (* p < 0.05, ** p < 0.01, student t test). (f) Quantification of the expressed VEGF protein on day 4 post dosage by ELISA.

Article Snippet: Culture supernatants (100 μL/well) were harvested and assayed for IFN‐α using the Mouse IFN‐α ELISA Kit (PBL Interferon Source) according to the manufacturer's instructions.

Techniques: Ligation, Confocal Laser Scanning Microscopy, Enzyme-linked Immunosorbent Assay

Journal: Bioactive Materials

Article Title: Splenic dendritic cell-targeting mRNA transfection of H-type ionizable lipid-based LNPs for enhancing tumor immunotherapy

doi: 10.1016/j.bioactmat.2026.02.018

Figure Lengend Snippet: In vitro effects of mOVA/H 18 NPs on activation of BMDCs. BMDCs were incubated with different formulations for 24 h and analyzed by flow cytometry. Quantification analysis for CD80 + CD86 + cells (A) and CD40 + cells (C) in BMDCs. Representative flow cytometry contour plots (B) for CD80 + CD86 + cells and histograms (D) for CD40 + cells in BMDCs. Concentrations of IL-4 (E), TNF-α (F), IFN-γ (G) and IL-12 (H) in BMDCs medium detected using ELISA. Data were shown as mean ± SD (n = 3).

Article Snippet: Mouse IFN-γ precoated ELISPOT kit was purchased from DAKEWE (Beijing, China).

Techniques: In Vitro, Activation Assay, Incubation, Flow Cytometry, Enzyme-linked Immunosorbent Assay

Journal: Bioactive Materials

Article Title: Splenic dendritic cell-targeting mRNA transfection of H-type ionizable lipid-based LNPs for enhancing tumor immunotherapy

doi: 10.1016/j.bioactmat.2026.02.018

Figure Lengend Snippet: In vivo DC maturation and T cell activation in C57BL/6J mice induced by mOVA/H 18 NPs through intravenous injection. C57BL/6J mice were vaccinated with different formulations on Day 0 and Day 5. On Day 10, the spleens of mice were collected and analyzed by flow cytometry. Quantification analysis of (A) CD80 + CD86 + DCs and (B) CD40 + DCs in the spleen. Quantification analysis of (C) CD3 + CD4 + T cells and (D) CD3 + CD8 + T cells in the spleen. (E) Quantification analysis and (F) representative flow cytometry contour plots of OVA-specific CD8 + T cells among all cell populations in the spleen. (G) Quantification analysis and (H) representative flow cytometry contour plots of IFN-γ + CD8 + T cells among all cell populations in the spleen. (I) Quantification results and (J) representative images of IFN- γ -secreting immune cells in the spleen of mice analyzed by enzyme-linked immunospot (ELISpot) assay. Data were shown as mean ± SD (n = 3).

Article Snippet: Mouse IFN-γ precoated ELISPOT kit was purchased from DAKEWE (Beijing, China).

Techniques: In Vivo, Activation Assay, Injection, Flow Cytometry, Enzyme-linked Immunospot

Journal: Bioactive Materials

Article Title: Splenic dendritic cell-targeting mRNA transfection of H-type ionizable lipid-based LNPs for enhancing tumor immunotherapy

doi: 10.1016/j.bioactmat.2026.02.018

Figure Lengend Snippet: Anti-tumor effects of mTrp2/H 18 NPs as therapeutic vaccines in vivo and inhibitory effects of mOVA/H 18 NPs on lung metastasis of B16-OVA. (A) Schematic illustration of experiment design. B16F10 cells were inoculated subcutaneously on C57BL/6J mice on Day 0. The B16F10 bearing mice were vaccinated on Day 5 and Day 10 through intravenous injection. On Day 16, the mice were sacrificed for further flow cytometry analysis. mTrp2/MC3-LNP was administered at a mTrp2 dose of 0.75 mg kg −1. As for mTrp2/H 18 NPs, the doses were set at 0.25 mg kg −1 for mTrp2/H 18 NPs (L), 0.5 mg kg −1 for mTrp2/H 18 NPs (M), and 0.75 mg kg −1 for mTrp2/H 18 NPs (H). (B) Tumor growth curves of B16F10-bearing mice after treatment with different formulations (n = 6). (C) Survival curves of B16F10-bearing mice treated with different formulations (n = 6). The survival rates of the two groups were analyzed using a log-rank test. Quantification analysis of IFN-γ + cells among CD3 + CD8 + T cells (D) in the spleen, (E) in tumor tissues, and (F) in the blood (n = 3).

Article Snippet: Mouse IFN-γ precoated ELISPOT kit was purchased from DAKEWE (Beijing, China).

Techniques: Vaccines, In Vivo, Injection, Flow Cytometry

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: Endothelial injury is a central driver of systemic IFN-β toxicity and is reversible through Jak inhibition

doi: 10.1016/j.omton.2026.201226

Figure Lengend Snippet: Elevated transaminases and toxicity from high plasma mIFNβ levels after hydrodynamic injection of plasmid DNA encoding mIFNβ in Balb/c mice (A) Schematic representation of the sleeping beauty transposon system with a transposon plasmid pKT2/mIFNβ-CLP PGK-Luc encoding mIFNβ, and sleeping beauty SB100X transposase. (B) Plasma IFN-β levels from Balb/c mice [0.0037 μg DNA ( n = 5), 0.011 μg DNA ( n = 3), 0.033 μg DNA ( n = 3), 0.05 μg DNA ( n = 3), 0.1 μg DNA ( n = 4), 0.2 μg DNA ( n = 3), 0.4 μg DNA ( n = 6), 0.8 μg DNA ( n = 3), and 1.6 μg DNA ( n = 1)] at 6–10 days after hydrodynamic injection of increasing amounts of plasmid DNA. Mean ± SEM. (C) AST and (D) ALT levels of individual mice with the corresponding plasma mIFNβ are plotted. The respective Pearson correlation factor and p values are shown. High systemic IFN-β is associated with elevated AST and ALT.

Article Snippet: Murine IFN-β was analyzed using VeriKine Mouse IFNβ ELISA kit (42400, PBL Assay Science, Piscataway, NJ).

Techniques: Clinical Proteomics, Injection, Plasmid Preparation

Journal: Molecular Therapy Oncology

Article Title: Endothelial injury is a central driver of systemic IFN-β toxicity and is reversible through Jak inhibition

doi: 10.1016/j.omton.2026.201226

Figure Lengend Snippet: Lack of toxicity in the absence of IFNR signaling and toxicity is reversible after IFN-β production is terminated Transgenic mice knockout for type I IFN receptor (IFNRko) and C57/B6 mice received one intramuscular injection of saline or 2 × 10 11 vg AAV1-mIFNβ and were monitored over time. (A) Plasma mIFNβ, (B) Kaplan-Meier survival curves, (C) ALT, (D) AST from saline (black, n = 6), or 2 × 10 11 vg AAV1-mIFNβ-treated IFNRko mice (red, n = 10), and C57B/6 mice (blue, n = 7) were plotted at regular intervals. Line denotes mean. Half of the mice were bled for each time point for blood analysis. C57/B6 mice ( n = 25) were given one intramuscular injection of AAV-mIFNβ-iCas9 and 14 days later, given one intraperitoneal dose of AP1903 a prodrug to activate iCas9 to initiate apoptosis of AAV-transduced muscle cells expressing IFN-β. (E) Plasma mIFNβ, (F) ALT, (G) AST, and (H) platelet counts in C57B/6 mice before (day 0) and after termination of mIFNβ production using AP1903. Mean ± SEM.

Article Snippet: Murine IFN-β was analyzed using VeriKine Mouse IFNβ ELISA kit (42400, PBL Assay Science, Piscataway, NJ).

Techniques: Transgenic Assay, Knock-Out, Injection, Saline, Clinical Proteomics, Expressing

Journal: Molecular Therapy Oncology

Article Title: Endothelial injury is a central driver of systemic IFN-β toxicity and is reversible through Jak inhibition

doi: 10.1016/j.omton.2026.201226

Figure Lengend Snippet: Concomitant treatment with ruxolitinib protects VSV-IFNβ-NIS treated MPC-11 tumor-bearing Balb/c mice from toxicity (A) Kaplan-Meier survival, (B) individual tumor volumes, (C) predicted tumor volumes using mixed effects modeling, (D) plasma IFNβ, (E) ALT, (F) AST, and (G) platelet count of mice at scheduled bleeds on day 3 and day 14, or at necropsy (at various time points). Treatment groups are color coded, saline (black, n = 5), VSV alone (red, n = 10), ruxolitinib (gray, n = 5), and VSV plus ruxolitinib (blue, n = 10). One intravenous dose of VSV at 10 7 TCID 50 was given per mouse, and/or ruxolitinib at 2 mg twice a day by oral gavage for 10 days, starting on the same day as VSV therapy.

Article Snippet: Murine IFN-β was analyzed using VeriKine Mouse IFNβ ELISA kit (42400, PBL Assay Science, Piscataway, NJ).

Techniques: Clinical Proteomics, Saline