mouse ifn-gamma elispot kit  (Bio-Techne corporation)

Journal: bioRxiv

Article Title: Coordinated Tuning of Ionizable Lipids and Formulation Redirects mRNA Vaccines Toward Lymphoid-Specific CD4 + T Cell Immunity

doi: 10.64898/2026.04.16.719092

Figure Lengend Snippet: (a) Vaccination scheme. BALB/c mice were immunized intramuscularly with SARS-CoV-2 spike mRNA-loaded LNPs (0.25 mg/kg) following a prime–boost regimen (day 0 and day 21). Blood samples and splenocytes were collected three weeks after the booster dose for antibody and T cell analysis. (b) Antigen-specific IgG levels at three weeks post-boost (n = 5 per group). (c) Neutralizing antibody titers assessed by PRNT₅₀ at three weeks post-boost (n = 3 per group). (d) Antigen-specific T cell responses assessed by IFN-γ ELISpot (n= 2–3 per group). (e) Viral challenge scheme. K18-hACE2 transgenic mice were immunized following the same prime–boost regimen and challenged with SARS-CoV-2 three weeks after the booster dose. (f) Body weight changes following viral challenge (n = 5 per group). (g) Survival curves following viral challenge (n = 5 per group). Statistical significance for (b–d) was determined by one-way ANOVA followed by Tukey’s multiple comparisons test. Survival curves were compared using the log-rank (Mantel-Cox) test. Data are presented as mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: The assay was performed using a mouse IFN-γ ELISpot kit (XEL485, R&D Systems) according to the manufacturer’s instructions.

Techniques: Cell Analysis, Enzyme-linked Immunospot, Transgenic Assay

Journal: Vaccines

Article Title: Peptide Coacervates Promote Cytosolic Delivery of STING Agonists for Cancer Immunotherapy

doi: 10.3390/vaccines14040329

Figure Lengend Snippet: Peptide and cGAMP coacervates vaccination induce epitope-specific T cell response in vivo. ( a ) Schematic illustration of the experimental design for peptide and cGAMP coacervates vaccination. C57BL/6 mice were subcutaneously injected with OVA-SIIN (40 nmol), SIIN-4A (40 nmol) and cGAMP (8 nmol) in indicated formulations at days 0, 7, and 14 at the tail base. ( b , c ) The percentage of SIINFEKL-specific CD8 + T cells (Tetramer + CD8 + T cells) ( b ) and statistics ( c ) in PBMCs at day 7 post-immunization after three immunizations ( n = 3). ( d , e ) IFN-γ ELISpot analysis ( d ) and spot statistics ( e ) of splenocytes (5 × 10 5 ) after re-stimulation with OVA peptide SIINFEKL at day 7 post-immunization after three immunizations ( n = 5). ( f , g ) C57BL/6 mice were subcutaneously injected with Cy5.5-OVA-SIIN (40 nmol), Cy5.5-SIIN-4A (40 nmol) or their combinatorial regimen with cGAMP (8 nmol) at the tail base for 24 h, then the inguinal lymph nodes were isolated and the fluorescent imaging were recorded by IVIS Spectrum imager ( f ) and the statistics of total photons ( g ). Data are presented as mean ± SD, n = 3 independent samples in ( b , c ); n = 5 independent samples in ( d , e ); n = 4 independent samples in ( f , g ), except Ctrl group ( n = 2); ** p = 0.0017, **** p < 0.0001 using one-way ANOVA with Tukey’s test ( c , e , g ).

Article Snippet: Mouse IFN-γ ELISpot kit (3321-4APT-2) was purchased from MABTECH (Stockholm, Sweden).

Techniques: In Vivo, Injection, Enzyme-linked Immunospot, Isolation, Imaging

Journal: eBioMedicine

Article Title: Nasal RNA-scaffold-protein vaccine protects mice from human H5N1 clade 2.3.4.4b virus lethal infection and safeguards against vaccine-unmatched viruses

doi: 10.1016/j.ebiom.2026.106228

Figure Lengend Snippet: Mutant proteins impacted RSPVac immunisation. (A) Schematic showing the disrupted formation of RSPVac using an RNA-binding-deficient mutant. Female BALF/c mice (n = 6) were nasally immunised with two doses of RSPVac-H5, 14-days apart. The RSPVac was produced with either the WT or mutant protein. Sera, BALF, and BALF-flushed cells were harvested 10 days post-2nd dose and analysed. (B) Fluorescence polarisation of WT and Mutant protein. After mutating major arginine residues, the RNA-binding affinity was reduced by 7-fold. (C–F) ELISA for anti-HA serum IgG, BALF IgA, Serum IgG subtypes, and BALF IgG subtypes. (G) IFNγ ELISpot analysis of BALF-flushed cells, stimulated with the immunogen (WT protein used to generate RSPVac-H5-1194). (A) Schematic of the phase separation mutant of the SARS2-RSPVac. To explore whether phase separation is related to RSPVac nasal immunisation, phase separation mutants (Psmut) were generated and used to generate SARS2-RSPVac. Female BALB/c mice were immunised using the same regimen with WT or Psmut RSPVac. Sera and BALF were harvested for downstream analysis. (B–F) ELISA for anti-spike RBD serum IgG, BALF IgG, BALF IgA, serum IgG1/IgG2a, BALF IgG1/IgG2a. (G) IFNγ ELISpot analysis of BALF-flushed cells, stimulated with the immunogen (protein used to generate WT SARS2-RSPVac). Statistical significance was determined by the Mann–Whitney test. p < 0.05 was considered significant. P-values were shown, and those that were considered statistically not significant were labelled not significant (ns). Figures A were created in BioRender.

Article Snippet: BALF cells were stimulated with recombinant proteins and IFN-secreting cells were detected by ELISpot Flex Mouse IFN-γ (ALP) kit (Mabtech Cat# 3321-2A).

Techniques: Mutagenesis, RNA Binding Assay, Produced, Fluorescence, Enzyme-linked Immunosorbent Assay, Enzyme-linked Immunospot, Generated, MANN-WHITNEY

Journal: eBioMedicine

Article Title: Nasal RNA-scaffold-protein vaccine protects mice from human H5N1 clade 2.3.4.4b virus lethal infection and safeguards against vaccine-unmatched viruses

doi: 10.1016/j.ebiom.2026.106228

Figure Lengend Snippet: Mouse antibody and T cell responses following RSPVac nasal vaccination. (A) Vaccination regimen. Female BALB/c mice (n = 6–9) were intranasally vaccinated with 2 doses of RSPVac, 14 days apart. Sera, BALF, and BALF cells were harvested at day 10 post-2nd dose for analysis. Control mice received the protein component without RNA (protein-only). Data was shown as box plots with respective colours indicating vaccine given, showing all values. Error bars represented the highest and lowest values of each box. ELISA was used to measure antibody responses for (B) anti-HA serum IgG, (C) anti-HA BALF IgG, (D) anti-HA BALF IgA, (E) anti-NP, and (F) serum anti-HA IgG subtypes for each of the RSPVac, respectively. (G) Mucosal T cell responses measured by IFNγ+ ELISpot using the immunogen (the protein used to generate RSPVac) stimulation of BALF cells. (H) To analyse lung T cells, female BALB/c mice (n = 6) were intranasally vaccinated with 2 doses of RSPVac, 14 days apart. Lungs were harvested at day 7 post-2nd dose. Single cell suspensions were prepared, and the NP peptide pool of H5N1, H1N1, and H7N9 was used to stimulate lung cells. Resident T cell responses were analysed by intracellular staining and FACS analysis. NP-reactive (I) CD4+ and (J) CD8+ T cells were shown, comparing protein-only and RSPVac-vaccinated. Statistical significance was determined by the Mann–Whitney test. p < 0.05 was considered significant. P-values were shown, and those that were considered insignificant were labelled not significant (ns). Figures A and H were created in BioRender.

Article Snippet: BALF cells were stimulated with recombinant proteins and IFN-secreting cells were detected by ELISpot Flex Mouse IFN-γ (ALP) kit (Mabtech Cat# 3321-2A).

Techniques: Control, Enzyme-linked Immunosorbent Assay, Enzyme-linked Immunospot, Single Cell, Staining, MANN-WHITNEY