mouse ifn γ elispot kit  (Cellular Technology Ltd)

Journal: Advanced Science

Article Title: Sustained Vaccine Exposure Elicits More Rapid, Consistent, and Broad Humoral Immune Responses to Multivalent Influenza Vaccines

doi: 10.1002/advs.202404498

Figure Lengend Snippet: Mechanistic investigation into breadth of humoral response provided by adjuvanted PNP‐hydrogel vaccines. a) Hemagglutination inhibition assay performed against homologous (H3 – A/Darwin/9/2021, B/Yamagata lineage – B/Phuket/3073/2013) or heterologous (H1 – A/Arkansas/08/2020, H3 – A/Tasmania/503/2020) influenza viruses on timepoint‐pooled sera. (n = 6) b) Schedule of vaccination with Fluzone formulations as described in Figure for additional immune assays. c) Homologous HA‐specific IFNγ‐producing T cells measured via splenocyte ELISpot at 3 weeks post vaccination. ELISpot well plate images show samples from one representative mouse in the Fluzone and PNP‐1‐5/3M‐052/Fluzone groups in wells coated with each of the peptide antigens. (n = 6) d) Schematic of influenza mAbs FluA‐20 and MEDI 8852 binding to head and stem regions, respectively, on HA. e) Competitive EILSA dilution curves of week 8 purified IgG against two mAbs, MEDI8852 (stem binder) and FluA‐20 (head binder), binding to A/Darwin/9/2021 HA. (n = 3, replicates of sera pooled from 9 mice) f) EC50 of MEDI dilution curves in (e). g) Area under the curve (AUC) of dilution curves in (e). Dotted lines indicate lower limit of detection. All error bars are mean ± SEM, P values determined by one‐way ANOVA with Tukey's multiple comparisons test (a) or a two‐tailed Student's t ‐test (c, f, g). Statistical significance was considered as P < 0.05.

Article Snippet: The frequency of antigen‐specific IFN‐γ producing splenocytes was determined by measuring antigen‐specific IFN‐γ producing CD8+ T cells using a Mouse IFN‐γ Single‐Color ELISPOT kit (CTL ImmunoSpot).

Techniques: Vaccines, HI Assay, Enzyme-linked Immunospot, Binding Assay, Purification, Two Tailed Test

Journal: Advanced Science

Article Title: Muco‐Penetrating Lipid Nanoparticles Having a Liquid Core for Enhanced Intranasal mRNA Delivery

doi: 10.1002/advs.202407383

Figure Lengend Snippet: Evaluation of mucosal immunity and inflammatory reaction following intranasal administration of iLLN‐2/mRNA complexes and ALC‐LNP. A) Illustration of prime‐boost intranasal immunization and downstream assay procedures. Created with BioRender.com. B) Levels of anti‐SARS‐CoV‐2 spike IgA and IgG in NLF. The dotted line indicates the limit of detection. C) IFN‐γ‐producing spot‐forming units (SFU) per 10 5 cells in the lung harvested on day 28. The dotted line indicates the limit of detection. Mean ± SD ( n = 3 for Ctrl, n = 5 for iLLN‐2/mRNA, n = 4 for ALC‐LNP). D) Kaplan–Meier survival curves of the immunized mice. E) Serum levels of albumin, total protein, and globulin on day 28. Mean ± SD ( n = 4–5); **** p < 0.0001; *** p < 0.001; ** p < 0.01 (one‐way ANOVA with Tukey's post hoc test). F) Serum levels of tumor necrosis factor‐α (TNF‐α) on day 7. Mean ± SD ( n = 8–10); * p < 0.05 (one‐way ANOVA with Tukey's post hoc test). G) H&E and Ly6G (neutrophil marker) staining of lung tissues harvested on day 28. Ly6G‐positive cells were marked by white arrowheads. The area labeled “F” indicates the fibrous tissue. Scale bar: 100 µm. H) Neutrophil counts in the lung were analyzed using ImageJ software. Mean ± SD ( n = 10); *** p < 0.001; ** p < 0.01 (one‐way ANOVA with Tukey's post hoc test).

Article Snippet: SARS‐CoV‐2 spike protein ELISA kit (GeneTex, USA), OptEIA mouse TNF ELISA kit (BD Biosciences, USA), and mouse interferon‐γ (IFN‐γ) single‐color ELISPOT kit (Cellular Technology Ltd, USA) were used as per manufacturer instructions.

Techniques: Marker, Staining, Labeling, Software