monkey ifn γ elispot pro kit  (Mabtech Inc)

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. 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: Human Mutation

Article Title: Whole‐Exome Sequencing to Screen Personal Neoantigens With High Immunogenicity in Patients With Microsatellite Stability (MSS)–Advanced Colorectal Cancer

doi: 10.1155/humu/3876230

Figure Lengend Snippet: Evaluating the immunogenicity of neoantigens from patients with CRC. Autologous PBMCs were stimulated with candidate mutated peptides every 3 days in the presence of IL‐2. On Day 10, T‐cell responses to each antigen were measured by an IFN‐ γ ELISpot assay. The PBMCs were obtained from P1 with CRC. No peptide (medium only) or VSV‐NP 43-69 (STKVALNDLRAYVYQGIKSGNPSILHI) stimulation was used as a control. Data are presented as mean ± SD of three independent experiments. ∗ p < 0.05 and ∗∗ p < 0.01 compared with IFN‐ γ production by PBMCs stimulated without peptide or with VSV‐NP 43-69 .

Article Snippet: After 10 days, the IFN‐ γ response from the prestimulated T cells against neoantigens was assessed using ELISpot assays with a Human IFN‐ γ ELISpot kit (Mabtech).

Techniques: Immunopeptidomics, Enzyme-linked Immunospot, Control

Journal: Human Mutation

Article Title: Whole‐Exome Sequencing to Screen Personal Neoantigens With High Immunogenicity in Patients With Microsatellite Stability (MSS)–Advanced Colorectal Cancer

doi: 10.1155/humu/3876230

Figure Lengend Snippet: Cytotoxicity of NRTs raised by in vitro stimulation of PBLs of P1. NRTs were induced with autologous ZNF169‐A275S– and CDH4‐V456M–pulsed DCs derived from the PBLs of P1. On Day 7, after the third stimulation, the NRTs were harvested for analysis. (A) IFN‐ γ secretion by NRT lines in response to mutated and WT peptides. IFN‐ γ ‐positive SFCs/10 5 NRTs were detected by cytokine‐specific ELISpot. (B–F) Cytotoxicity at the indicated E:T ratios measured by a CCK‐8 kit. Peptide‐specific targets were mutated protein–pulsed T2 cells and minimally nucleated SW480 cells, whereas VSV‐NP 43-69 –pulsed T2 cells, T2 cells alone, and SW480 cells alone were used as controls. Data are expressed as mean ± SEM ( ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001).

Article Snippet: After 10 days, the IFN‐ γ response from the prestimulated T cells against neoantigens was assessed using ELISpot assays with a Human IFN‐ γ ELISpot kit (Mabtech).

Techniques: In Vitro, Derivative Assay, Enzyme-linked Immunospot, CCK-8 Assay