pre coated ifn γ  (Mabtech Inc)

Journal: International Journal of Pharmaceutics: X

Article Title: Engineering a safe and potent LNP-mRNA delivery system by leveraging the dual activities of α-tocopherol

doi: 10.1016/j.ijpx.2026.100553

Figure Lengend Snippet: Humoral and cellular immunogenicity. (A) Timeline of vaccination and sample collection. (B) The RSV pre-F protein-specific IgG antibody levels of modified LNP-mRNA complexes after the boost immunization, as detected by ELISA. (C) The RSV pre-F protein-specific IgG1 and IgG2a antibody levels and the corresponding ratio of LogIgG2a/LogIgG1 of modified LNP-mRNA complexes after the boost immunization, as detected by ELISA. (D) The neutralizing antibody titers against live virus across all treatment groups. (E) The quantification of restimulated IFN-γ-secreting and IL-17 A-secreting splenocytes was verified in the FluoroSpot test. (F) Results of TEM and TCM in splenic CD4 + T cells. (G) The percentage of naïve CD4 + T cells and CD8 + T cells. (H) Results of TEM and TCM in splenic CD8 + T cells. Data are presented as mean ± SEM. Statistical significance was determined using one-way ANOVA followed by Dunnett's multiple comparisons test, with all comparisons made against the Std-LNP group. All data showed no statistically significant differences and were not labeled in the figure.

Article Snippet: IFN-γ and IL-17 A FluoroSpot assay: To quantify antigen-specific T cells, splenocytes (2.5 × 10 5 cells/well) were stimulated with an overlapping peptide pool (length = 15 aa, overlap = 10 aa) spanning the RSV pre-F protein (2 μg/mL per peptide) in pre-coated IFN-γ and IL-17 A FluoroSpot plates (Mabtech, Sweden).

Techniques: Immunopeptidomics, Modification, Enzyme-linked Immunosorbent Assay, Virus, Labeling

Journal: Journal of Neuroinflammation

Article Title: Sez6L2 autoimmunity induces cerebellar ataxia in mice

doi: 10.1186/s12974-025-03637-7

Figure Lengend Snippet: Sez6L2 immunized mice generate Sez6L2-specific T cells. A and B Splenocyte and lymph node mixed cell cultures from Sez6L2 or sham immunized mice were left unstimulated or were stimulated with H-Sez6L2 or M-Sez6L2 protein as indicated for 24 h in an IFNγ ELISPOT assay. N = 8–10 mice (4–5 male and 4–5 female); one mouse = one culture. Cells from mice immunized with M-Sez6L2 were only stimulated with M-Sez6L2 (not H-Sez6L2) as only the reaction to M-Sez6L2 is relevant to the autoimmunity model for that group. A Representative images of ELISPOT wells. B Quantification of the number of spots per 200,000 splenocytes/lymph node cells plated. C - E Splenocyte/lymph node mixed cell cultures from Sez6L2 or sham immunized mice were stimulated with M-Sez6L2 protein for 24–72 h followed by cell surface and intracellular cytokine staining and analysis by flow cytometry. C Graphs show the percent of CD4 + cells positive for IFNγ or TNFα after stimulation in culture for 24 h. N = 8–10 (4–5 male and 4–5 female). D Graphs show the percent of CD4 + cells positive for IFNγ, TNFα, IL-4, or IL-17 A after stimulation in culture for 72 h. N = 4–5 males only. E Graphs show the percent of CD8 + cells positive for IFNγ, TNFα after stimulation in culture for 24 h. N = 8–10 (4–5 male and 4–5 female). Statistics for all graphs = Welch-ANOVAs with Dunnett’s T3 MCTs

Article Snippet: Mixed splenocyte and lymph node cells were plated on IFNγ ELISPOT pre-coated plates (Cellular Technology Limited, mIFNgp-2 M) in serum-free media at a cell density of 200,000 cells per well with or without 8 μg/mL mouse or human recombinant Sez6L2 protein.

Techniques: Enzyme-linked Immunospot, Staining, Flow Cytometry

Journal: Journal of Neuroinflammation

Article Title: Sez6L2 autoimmunity induces cerebellar ataxia in mice

doi: 10.1186/s12974-025-03637-7

Figure Lengend Snippet: Immunodominant T cell epitopes and antibody peptide epitopes from Sez6L2-immunized mice. A Splenocytes from H-Sez6L2 or sham-immunized mice were stimulated with 15-mer/10aa overlapping peptide library containing peptides from the extracellular domain of mouse and human Sez6L2 and assayed in an IFNγ ELISPOT assay. The 15-mer immunodominant peptide sequences are shown with the MHC-II core binding region predicted by IEDB Resource highlighted in red. For the peptide ELISPOT, H-Sez6L2-immunized N=5 (3 male, 2 female). B ELISA absorbance from serum of 6-week post-immunization mice tested for binding to the Sez6L2 15-mer peptide library. 12 H-Sez6L2-immunized mice were tested. For both A and B) Circle data points represent results from peptides that are identical in human and mouse Sez6L2, diamond data points are from peptides unique to humans, and triangle data points are from peptides unique to mouse. The colors represent individual mice. A schematic of Sez6L2’s domain structure is aligned to the A and B graphs. The red lines highlight the location of immunodominant T cell epitopes and the green lines highlight the antibody epitopes. C Summary table from 15-mer experiments showing peptide sequences that had positive hits in the ELISPOT and/or ELISA experiments. MHC-II core binding region predicted by IEDB Resource are highlighted in red. Overlapping peptides sharing 10 amino acids in common are underlined.

Article Snippet: Mixed splenocyte and lymph node cells were plated on IFNγ ELISPOT pre-coated plates (Cellular Technology Limited, mIFNgp-2 M) in serum-free media at a cell density of 200,000 cells per well with or without 8 μg/mL mouse or human recombinant Sez6L2 protein.

Techniques: Enzyme-linked Immunospot, Binding Assay, Enzyme-linked Immunosorbent Assay