mouse ifn γ elispot basic kit  (Mabtech Inc)

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. Nucleic Acids

Article Title: Comparative analysis of expression, immunogenicity, and safety profiles between linear and circular RNA vaccine platforms

doi: 10.1016/j.omtn.2026.102954

Figure Lengend Snippet: Cell-mediated immune responses to different RNA platforms Each group includes a negative control (DPBS), unmodified linear mRNA (Lin-WT), modified linear mRNA (Lin-m1Ψ), and unmodified circular RNA (Circ-WT). (A) Immunization schedule schematic diagram for assessing T cell responses. Balb/c mice were intramuscularly immunized twice, 2 weeks apart, with 10 μg of LNP-encapsulated HA-encoding mRNA (Lin-WT, Lin-m1Ψ, or Circ-WT). (B and C) IFN-γ ELISPOT assay showing the number of antigen-specific IFN-γ-secreting splenocytes following immunization with HA-encoding mRNA. (D–F) Flow cytometry analysis of cytokine-producing CD8+ T cells. The frequencies of IFN-γ+, IL-2+, and TNF-α + CD8+ T cells were assessed to evaluate antigen-specific T cell activation. (G–I) Cytokine-producing CD4+ T cells, with frequencies of IFN-γ+, IL-2+, and TNF-α+ CD4+ T cells measured by intracellular cytokine staining. (J) Analysis of double-positive cytokine-expressing CD8+ T cells, indicating polyfunctional T cell responses. (K) Analysis of double-positive cytokine-expressing CD4+ T cells, indicating helper T cell activation. Data represent mean ± SD ( n = 5 per group). Statistical significance was determined by ordinary one-way ANOVA with Tukey’s test or the Kruskal-Wallis test with Dunn’s multiple comparison test, depending on the normality of the data. ∗ p ≤ 0.05, ∗∗ p ≤ 0.01, ∗∗∗∗ p ≤ 0.0001.

Article Snippet: IFN-γ secreting T cells were detected using the ELISpot assay and the mouse IFN-γ ELISpot BASIC kit from Mabtech (Stockholm, Sweden), following the manufacturer’s instructions.

Techniques: Negative Control, Modification, Enzyme-linked Immunospot, Flow Cytometry, Activation Assay, Staining, Expressing, Comparison

Journal: Molecular Therapy. Nucleic Acids

Article Title: Comparative analysis of expression, immunogenicity, and safety profiles between linear and circular RNA vaccine platforms

doi: 10.1016/j.omtn.2026.102954

Figure Lengend Snippet: Differential innate immune responses induced by different RNA platforms Groups comprise the negative control (DPBS), unmodified linear mRNA (Lin-WT), modified linear mRNA (Lin-m1Ψ), and unmodified circular RNA (Circ-WT). (A) Schematic presentation of in vivo cytokine analysis. Mice were injected intramuscularly with LNP-encapsulated mRNA, and cytokine levels were assessed in serum and lymph nodes at 6- and 24-h post-inoculation. (B–G) In vivo cytokine responses measured in serum and lymph nodes at different time points. Levels of (B-C) IFN-γ, (D) RIG-I, (E) IFN-β, (F) TNF-α, (G) IL-6. mRNA Fold change calculated by the 2 −ΔΔCt method and normalized to GAPDH, with values expressed as fold change relative to the DPBS group. Data represent mean ± SD ( n = 5 per group). Statistical significance was determined by ordinary one-way ANOVA with Tukey’s test or the Kruskal-Wallis test with Dunn’s multiple comparison test, depending on the normality of the data. ∗ p ≤ 0.05, ∗∗ p ≤ 0.01, ∗∗∗ p ≤ 0.001, ∗∗∗∗ p ≤ 0.0001.

Article Snippet: IFN-γ secreting T cells were detected using the ELISpot assay and the mouse IFN-γ ELISpot BASIC kit from Mabtech (Stockholm, Sweden), following the manufacturer’s instructions.

Techniques: Negative Control, Modification, In Vivo, Injection, Comparison

Journal: Virus Research

Article Title: Development and protective evaluation of mRNA vaccines encoding the glycoprotein precursor of lymphocytic choriomeningitis virus

doi: 10.1016/j.virusres.2026.199735

Figure Lengend Snippet: Cellular immune responses mapped by peptide pools. A IFN-γ ELISpot responses of splenocytes collected at Day 45 post-prime from BALB/c mice immunized with Arm-Vax, Cl13-Vax, the bivalent Arm+Cl13-Vax, or mock control. Cells were restimulated ex vivo with the indicated GPC peptide pools (Shared, Arm-full, Cl13-full, Arm-only, and Cl13-only), and responses are presented as spot-forming units (SFUs) per 10 6 splenocytes. B Representative ELISpot well images showing responses to a positive control stimulus (PHA), the Shared peptide pool, and a negative control for each immunization group.

Article Snippet: The frequency of antigen-specific IFN-γ-secreting splenocytes was determined using the Mouse IFN-γ Precoated ELISpot Kit (Mabtech, Sweden; Cat# 3321–4HST) according to the manufacturer’s instructions.

Techniques: Enzyme-linked Immunospot, Control, Ex Vivo, Positive Control, Negative Control

Journal: Redox Biology

Article Title: A novel photosensitizer-based photodynamic therapy reprograms the Kynurenine–AhR axis to boost antitumor immunity in breast cancer

doi: 10.1016/j.redox.2026.104171

Figure Lengend Snippet: DTP-PDT attenuates the IDO–Kyn–AhR axis and relieves immune-suppression in the TME. (A-D) Targeted metabolomics analysis of Trp, Kyn, QA, and 5-HT in cell samples. (E) Targeted metabolomics of Kyn in tumor tissue samples. (F) Levels of Kyn in culture supernatants after the indicated various treatments under IFN-γ priming (n = 3). (G) Representative immunofluorescence images of tumor sections stained for CD3 (green), AhR (red), and DAPI (blue). Scale bar = 20 μm. (H) RT-qPCR analysis of Cyp1a1, Cyp1b1, and Ahrr mRNA expression in tumor-infiltrating CD3 + T cells (n = 4). (I-J) Proportion of intratumoral CD8 + T cells (gated on CD3 + T cells, n = 5). (K-L) Proportion of intratumoral Treg cells (gated on CD3 + CD4 + Foxp3 + T cells, n = 5). (M) Representative immunofluorescence images of tumor sections stained for CD3 (green), AhR (red), and DAPI (blue) in the Kyn rescue experiment. Scale bar = 20 μm. (N) RT-qPCR analysis of Cyp1a1, Cyp1b1, and Ahrr mRNA expression in tumor-infiltrating CD3 + T cells from the Kyn rescue experiment (n = 4). (O–P) Proportion of intratumoral CD8 + T cells in the Kyn rescue experiment (gated on CD3 + T cells, n = 5). (Q-R) Proportion of intratumoral Treg cells in the Kyn rescue experiment (gated on CD3 + CD4 + Foxp3 + T cells, n = 5). Data are shown as mean ± SD. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: 4T1 cells (5 × 10 5 cells/well) were seeded in 6-well plates and allowed to adhere for 24 h. To activate IDO1 activity, cells were pretreated with recombinant mouse interferon-γ (IFN-γ) (50 ng/mL, 485-MI, R&D System) for 24 h. After IFN-γ priming, cells were treated with DTP-PDT and/or the IDO1 inhibitor (NLG919, Aladdin).

Techniques: Immunofluorescence, Staining, Quantitative RT-PCR, Expressing