themouse ifn g elispot kit  (R&D Systems)

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