Journal: bioRxiv

Article Title: Immunogenicity and protective efficacy of an intranasal neuraminidase-based influenza virus vaccine adjuvanted with bacterial cell membrane-derived adjuvants

doi: 10.1101/2025.02.26.640278

Figure Lengend Snippet: a. Representative flow cytometry plots showing the percentage of IL-2- and TNFα-producing CD4 + EM T-lymphocytes in mouse lungs and spleens 10 days after the booster immunization (gated on live CD3 + CD19 ‒ CD4 + CD62L ‒ CD44 + cells). Cells were incubated with overlapping peptides, covering the whole sequence of N1 NA-protein for 6h in the presence of Brefeldin A and co-stimulatory anti-CD28 antibodies. b. Percentage of different cytokine-producing cell populations within the total CD4 + EM T-cell subset after background subtraction. Average and SE values are shown. Statistical analyses were performed using one-way ANOVA with Tukey posthoc test. P-values for the groups with statistically significant differences are shown on the plots.

Article Snippet: For the analysis of T- and B-cell populations in lungs, spleen, and NALTs, cells were stained with the antibody cocktail, including the following antibodies: CD38-AF488 (0.125 μl, BioLegend), PD-1-PE/Dazzle (0.5 μl, BioLegend), CD273-RB744 (0.25 μl, BD Biosciences), CD138-PE (0.25 μl, BioLegend), CD103-PE-Cy5.5 (0.5 μl, BioLegend), CD44-PE/Cy7 (0.25 μl, Tonbo Biosciences), GL7-APC (0.25 μl, BioLegend), CD69-AF700 (1 μl, BioLegend), CD62L-APC/Cy7 (0.125 μl, BioLegend), CD185-BV421 (0.5 μl, BioLegend), CD80-BV605 (1 μl, BioLegend), IgD-BV650 (0.25 μl, BioLegend), CD3-BV711 (0.5 μl, BioLegend), CD8-BV785 (0.25 μl, BioLegend), CD4-BUV496 (0.125 μl, BD Biosciences), IgM-BUV737 (1 μl, BD Biosciences), CD19-BUV805 (0.125 μl, BD Biosciences).

Techniques: Flow Cytometry, Incubation, Sequencing

Journal: bioRxiv

Article Title: Immunogenicity and protective efficacy of an intranasal neuraminidase-based influenza virus vaccine adjuvanted with bacterial cell membrane-derived adjuvants

doi: 10.1101/2025.02.26.640278

Figure Lengend Snippet: a. Body weight dynamics and survival after the heterologous challenge with 5 × LD 50 of A/bald eagle/Florida/W22-134-OP/2022 (H5N1 reassortant with A/Puerto Rico/8/1934 vaccine backbone) and A/New Caledonia/20/1999 (H1N1) (n = 8). b. Viral titers in lungs and nasal turbinates on day 4 after the challenge (n = 4). c. Representative flow cytometry plots showing the percentage of IL-2- and TNFα-producing CD4 + EM T-lymphocytes (gated on live CD3 + CD19 ‒ CD4 + CD62L ‒ CD44 + cells) in mouse lungs and spleens 5 days after the challenge with 0.1 × LD 50 of A/bald eagle/Florida/W22-134-OP/2022 (H5N1). Cells were incubated with overlapping peptides, covering the whole sequence of N1 NA-protein for 6h in the presence of Brefeldin A and co-stimulatory anti-CD28 antibodies. d. Percentage of different cytokine-producing cell populations within the total CD4 + EM T-cell subset after background subtraction. Average and SE values are shown. Statistical analyses were performed using one-way ANOVA with Tukey posthoc test. P-values for the groups with statistically significant differences are shown on the plots.

Article Snippet: For the analysis of T- and B-cell populations in lungs, spleen, and NALTs, cells were stained with the antibody cocktail, including the following antibodies: CD38-AF488 (0.125 μl, BioLegend), PD-1-PE/Dazzle (0.5 μl, BioLegend), CD273-RB744 (0.25 μl, BD Biosciences), CD138-PE (0.25 μl, BioLegend), CD103-PE-Cy5.5 (0.5 μl, BioLegend), CD44-PE/Cy7 (0.25 μl, Tonbo Biosciences), GL7-APC (0.25 μl, BioLegend), CD69-AF700 (1 μl, BioLegend), CD62L-APC/Cy7 (0.125 μl, BioLegend), CD185-BV421 (0.5 μl, BioLegend), CD80-BV605 (1 μl, BioLegend), IgD-BV650 (0.25 μl, BioLegend), CD3-BV711 (0.5 μl, BioLegend), CD8-BV785 (0.25 μl, BioLegend), CD4-BUV496 (0.125 μl, BD Biosciences), IgM-BUV737 (1 μl, BD Biosciences), CD19-BUV805 (0.125 μl, BD Biosciences).

Techniques: Flow Cytometry, Incubation, Sequencing