Journal: Nature Communications
Article Title: A vaccine platform targeting lung-resident memory CD4 + T-cells provides protection against heterosubtypic influenza infections in mice and ferrets
doi: 10.1038/s41467-024-54620-4
Figure Lengend Snippet: a , b WT (n = 5) and IFNAR1 −/− mice (n = 5) were immunized i.n . with NVT + H3N2. One day later, lung lysates and serum were collected. a Pooled lung lysates and serum samples were analyzed using a mouse cytokine kit. The signals were normalized to the pixel density of the indicated reference spots. b Concentrations of CCL2, CCL5, CXCL9, and CXCL10 in lung lysates and sera were measured via ELISA. c BALB/c Mice (n = 5 per group) were immunized i.n . with H3N2 or NVT. One day later, the levels of CCL2, CCL5, CXCL9, and CXCL10 in lung lysates and sera were assessed via ELISA. b , c Data are expressed as dot plots, with horizontal lines representing the medians. The two-sided Mann–Whitney U test was used for statistical analysis of differences between two groups. d , e BALB/c mice were immunized i . m . with H3N2 (n = 20), H3N2 + NVT (n = 10), or i . n . with H3N2 + NVT (n = 10) twice at two-week intervals. Two weeks later, mice injected i . m . with H3N2 (n = 10) or H3N2 + NVT (n = 10) were boosted i.n . with NVT. d On day 42, the proportion of lung CD4 + T RM cells from vaccinated mice (n = 5 per group) was determined via flow cytometry. e The other vaccinated mice (n = 5 per group) were infected i.n . with 4 LD 50 of H1N1 virus, and survival was monitored daily. Survival data are represented as Kaplan‒Meier survival curves, and the significant differences in survival rates were calculated by the log-rank test. f BALB/c mice were immunized i.n . with H3N2 + NVT (n = 15) or i.m . with H3N2 (n = 20). Two weeks later, the mice were immunized i.m . with H3N2 or i.n . with H3N2 + NVT, H3N2, or NVT, and the proportion of lung CD4 + T RM cells was determined via flow cytometry on day 42. d , f Box and whisker plots showing the median (center), 25th and 75th percentiles (box), and lowest and highest values (whiskers). Statistical analyses were performed using one-way ANOVA with the Tukey’s multiple comparisons test. This study was performed once. ns, not significant. Source data are provided as a Source Data file.
Article Snippet: Relative levels of cytokines and chemokines in lung lysates and serum from vaccinated mice were determined by a Proteome Profiler Mouse Cytokine Array Kit (Panel A, R&D Systems, Minneapolis, MN, USA) in accordance with the manufacturer’s recommendations.
Techniques: Enzyme-linked Immunosorbent Assay, MANN-WHITNEY, Injection, Flow Cytometry, Infection, Virus, Whisker Assay
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