Journal: Human Vaccines & Immunotherapeutics

Article Title: Intranasal immunization with a single dose of the fusion protein formulated with a combination adjuvant induces long-term protective immunity against respiratory syncytial virus

doi: 10.1080/21645515.2017.1349584

Figure Lengend Snippet: RSV ΔF-specific systemic humoral immune responses in mice. (A) Serum IgG1 and IgG2a titers before challenge (B/C), (B) Serum IgG1 and IgG2a titers after challenge (A/C) with RSV, (C) Serum VN antibody titers determined before (B/C) and after (A/C) RSV challenge, and (D) Numbers of IFN-γ and IL-5 secreting splenocytes determined in response to in vitro restimulation with ΔF protein. BALB/c mice were immunized once IN with ΔF formulated with TriAdj and challenged 3 weeks later with RSV. Control groups were immunized with PBS and challenged with RSV (Placebo) or mock-challenged (Placebo/mock). ELISA titers are expressed as the reciprocal of the highest dilution resulting in a value of two standard deviations above the negative control serum. Virus neutralization titers are expressed as the highest dilution of serum that resulted in <50% of cells displaying cytopathic effects. Cytokine secreting cell numbers are expressed as the difference in the number of spots between ΔF-stimulated wells and medium-control wells. Bars indicate median values with interquartile ranges. **P < 0.01; ***P < 0.001.

Article Snippet: Briefly, splenocytes and LN cells were stimulated with ΔF protein (2 µg/ml) and spots were developed with biotinylated IFN-γ- or IL-5-specific goat anti-mouse IgG (BD Biosciences, 554410, 554397) or biotinylated goat anti-mouse IgA-specific antibody, followed by AP-conjugated streptavidin and BCIP/NBT (Sigma-Aldrich, B5655) as the substrate.

Techniques: In Vitro, Enzyme-linked Immunosorbent Assay, Negative Control, Neutralization

Journal: Human Vaccines & Immunotherapeutics

Article Title: Intranasal immunization with a single dose of the fusion protein formulated with a combination adjuvant induces long-term protective immunity against respiratory syncytial virus

doi: 10.1080/21645515.2017.1349584

Figure Lengend Snippet: Mucosal immune responses to RSV ΔF protein in mice after challenge with RSV. The IgA titers (A), virus titers (B), percentages of ΔF-specific CD8 + T cells (C), and ΔF-specific IFN-γ secreting CD8 + T cells (D) were measured in the lung after RSV challenge. Mice were immunized and challenged as described in the legend for Fig. 1 . ELISA titers are expressed as the reciprocal of the highest dilution resulting in a value of two standard deviations above the negative control serum. Virus replication in the lungs is expressed as pfu per gram of lung tissue. The proportion of RSV F-specific CD8 + T cells was determined in the lung by KYKNAVTEL-MHC I pentamer staining. IFN-γ expression by CD8 + T cells was determined by intracellular cytokine staining. Bars represent median values with interquartile ranges. *P < 0.05; **P < 0.01; ***P < 0.001.

Article Snippet: Briefly, splenocytes and LN cells were stimulated with ΔF protein (2 µg/ml) and spots were developed with biotinylated IFN-γ- or IL-5-specific goat anti-mouse IgG (BD Biosciences, 554410, 554397) or biotinylated goat anti-mouse IgA-specific antibody, followed by AP-conjugated streptavidin and BCIP/NBT (Sigma-Aldrich, B5655) as the substrate.

Techniques: Enzyme-linked Immunosorbent Assay, Negative Control, Staining, Expressing

Journal: Human Vaccines & Immunotherapeutics

Article Title: Intranasal immunization with a single dose of the fusion protein formulated with a combination adjuvant induces long-term protective immunity against respiratory syncytial virus

doi: 10.1080/21645515.2017.1349584

Figure Lengend Snippet: Systemic immune responses to RSV ΔF protein. (A) Serum IgG1 and IgG2a titers before challenge (B/C), (B) Serum IgG1 and IgG2a titers after RSV challenge (A/C), (C) Serum VN antibody titers after RSV challenge (A/C), and (D) Numbers of IFN-γ and IL-5 secreting splenocytes determined in response to in vitro restimulation with ΔF protein. BALB/c mice were immunized once IN with ΔF formulated with TriAdj and challenged with RSV on day 150. Control groups were immunized with PBS and challenged with RSV (Placebo) or mock-challenged (Placebo/mock). ELISA titers are expressed as the reciprocal of the highest dilution resulting in a value of two standard deviations above the negative control serum. Virus neutralization titers are expressed as the highest dilution of serum that resulted in <50% of cells displaying cytopathic effects. Cytokine secreting cell numbers are expressed as the difference in the number of spots between ΔF-stimulated wells and medium-control wells. Bars indicate median values with interquartile ranges. ***P < 0.001.

Article Snippet: Briefly, splenocytes and LN cells were stimulated with ΔF protein (2 µg/ml) and spots were developed with biotinylated IFN-γ- or IL-5-specific goat anti-mouse IgG (BD Biosciences, 554410, 554397) or biotinylated goat anti-mouse IgA-specific antibody, followed by AP-conjugated streptavidin and BCIP/NBT (Sigma-Aldrich, B5655) as the substrate.

Techniques: In Vitro, Enzyme-linked Immunosorbent Assay, Negative Control, Neutralization