mouse ifn γ elispot basic kit  (Mabtech Inc)

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: iScience

Article Title: Incorporation of noncanonical base Z yields modified mRNA with minimal immunogenicity and improved translational capacity in mammalian cells

doi: 10.1016/j.isci.2023.107739

Figure Lengend Snippet: Development and evaluation of Z-mRNA-based COVID-19 vaccine in vivo (A) Schematic representation of the A-to-Z substitution in Z-mRNA and its advantages over unmodified synthetic mRNA. (B) Synthesis of mRNA LNPs via microfluidic mixing. Ionizable lipid: D-Lin-MC3-DMA. DSPC: distearoylphosphatidylcholine. DMG-PEG2000: 1,2-dimyristoyl-rac-glycero-3-methoxypolyethylene glycol-2000. (C) Characterization of Z- and m1ψ-mRNA LNPs. EE: encapsulation efficiency (top left). PDI: polydispersity index (top left). The size distribution was detected by dynamic light scattering and the average of three technical replicates was shown in the figure. (D) Mouse vaccination scheme adopted in this study. (E) SARS-CoV-2 RBD-specific IgG antibodies in sera determined by ELISA. Mice sera was collected 7 days or 14 days after second immunization and used for RBD-specific IgG antibody detection by ELISA. N = 6. Error bars represent means ± SEM. p value was calculated by two-tailed unpaired t test. (F) Pseudovirus neutralization curve. Mice sera collected 14 days after second immunization was used for pseudovirus neutralization assay. N = 6. Error bars represent means ± SEM. Dashed line: 0.5. (G) Representative well images of ELISPOT assays. Mouse splenocytes were isolated 35 days after second immunization for ELISPOT assay. Image here shows the spot forming units (based on IL-2 or IFN-γ secretion) from mouse splenocytes stimulated in vitro with the antigen RBD peptide-mix pool. (H and I) ELISPOT assays of IL-2 (H) and IFN-γ (I) secretion. Error bars represent means ± SEM. N = 6 × 3 = 18 in (H) and (I) (three ELISPOT replicates per mouse splenocyte sample). p value was calculated by two-tailed unpaired t test, ∗: p < 0.05, ∗∗: p < 0.01, ∗∗∗: p < 0.001, ∗∗∗∗: p < 0.0001. n.s. = not significant. See also Figures S10–S12 .

Article Snippet: ELISPOT assays were performed with Mabtech Mouse IL-2 ELISpot (plus) Kit (#3441-4HPW-2) and Mabtech Mouse IFN-γ ELISpot (basic) Kit (#3321-2H) following manufacturer’s protocols.

Techniques: In Vivo, Encapsulation, Enzyme-linked Immunosorbent Assay, Two Tailed Test, Neutralization, Enzyme-linked Immunospot, Isolation, In Vitro

Journal: iScience

Article Title: Incorporation of noncanonical base Z yields modified mRNA with minimal immunogenicity and improved translational capacity in mammalian cells

doi: 10.1016/j.isci.2023.107739

Figure Lengend Snippet:

Article Snippet: ELISPOT assays were performed with Mabtech Mouse IL-2 ELISpot (plus) Kit (#3441-4HPW-2) and Mabtech Mouse IFN-γ ELISpot (basic) Kit (#3321-2H) following manufacturer’s protocols.

Techniques: Virus, Isolation, Recombinant, Enzyme-linked Immunosorbent Assay, Enzyme-linked Immunospot, Software

Journal: iScience

Article Title: Incorporation of noncanonical base Z yields modified mRNA with minimal immunogenicity and improved translational capacity in mammalian cells

doi: 10.1016/j.isci.2023.107739

Figure Lengend Snippet: Development and evaluation of Z-mRNA-based COVID-19 vaccine in vivo (A) Schematic representation of the A-to-Z substitution in Z-mRNA and its advantages over unmodified synthetic mRNA. (B) Synthesis of mRNA LNPs via microfluidic mixing. Ionizable lipid: D-Lin-MC3-DMA. DSPC: distearoylphosphatidylcholine. DMG-PEG2000: 1,2-dimyristoyl-rac-glycero-3-methoxypolyethylene glycol-2000. (C) Characterization of Z- and m1ψ-mRNA LNPs. EE: encapsulation efficiency (top left). PDI: polydispersity index (top left). The size distribution was detected by dynamic light scattering and the average of three technical replicates was shown in the figure. (D) Mouse vaccination scheme adopted in this study. (E) SARS-CoV-2 RBD-specific IgG antibodies in sera determined by ELISA. Mice sera was collected 7 days or 14 days after second immunization and used for RBD-specific IgG antibody detection by ELISA. N = 6. Error bars represent means ± SEM. p value was calculated by two-tailed unpaired t test. (F) Pseudovirus neutralization curve. Mice sera collected 14 days after second immunization was used for pseudovirus neutralization assay. N = 6. Error bars represent means ± SEM. Dashed line: 0.5. (G) Representative well images of ELISPOT assays. Mouse splenocytes were isolated 35 days after second immunization for ELISPOT assay. Image here shows the spot forming units (based on IL-2 or IFN-γ secretion) from mouse splenocytes stimulated in vitro with the antigen RBD peptide-mix pool. (H and I) ELISPOT assays of IL-2 (H) and IFN-γ (I) secretion. Error bars represent means ± SEM. N = 6 × 3 = 18 in (H) and (I) (three ELISPOT replicates per mouse splenocyte sample). p value was calculated by two-tailed unpaired t test, ∗: p < 0.05, ∗∗: p < 0.01, ∗∗∗: p < 0.001, ∗∗∗∗: p < 0.0001. n.s. = not significant. See also Figures S10–S12 .

Article Snippet: Mouse IFN-γ ELISpot (basic) Kit , Mabtech , Cat#3321-2H.

Techniques: In Vivo, Encapsulation, Enzyme-linked Immunosorbent Assay, Two Tailed Test, Neutralization, Enzyme-linked Immunospot, Isolation, In Vitro

Journal: iScience

Article Title: Incorporation of noncanonical base Z yields modified mRNA with minimal immunogenicity and improved translational capacity in mammalian cells

doi: 10.1016/j.isci.2023.107739

Figure Lengend Snippet:

Article Snippet: Mouse IFN-γ ELISpot (basic) Kit , Mabtech , Cat#3321-2H.

Techniques: Virus, Isolation, Recombinant, Enzyme-linked Immunosorbent Assay, Enzyme-linked Immunospot, Software