Journal: International Archives of Allergy and Immunology

Article Title: Importance of NK Cells in Cellular and Humoral Responses Triggered by Pneumococcus Vaccination

doi: 10.1159/000535562

Figure Lengend Snippet: Gating strategy to identify T H populations of interest. Lymphocyte populations were gated based on FSC/SSC, excluding aggregates. T H cells were then gated from CD3+CD4+ cells, and T H 1, T H 2, and T H 17 cell populations were identified based on IFN-γ, IL-4, and IL-17 positivity, respectively.

Article Snippet: Of those cells, 5 × 10 6 were removed and added into 15 mL falcon tubes for flow cytometry analysis of CD4+ T-cell subset levels after 24-h treatment with 5 μg/mL PPV-23 vaccine antigen and addition of 5 μg/mL brefeldin A (BioLegend, 420601) at 16th hour of incubation.

Techniques:

Journal: International Archives of Allergy and Immunology

Article Title: Importance of NK Cells in Cellular and Humoral Responses Triggered by Pneumococcus Vaccination

doi: 10.1159/000535562

Figure Lengend Snippet: Comparison of CD4+ T-cell subset levels. Splenocytes collected 4 weeks after the PPV-23 vaccination were stimulated by PPV-23 vaccine antigen for 24 h. Antibodies against IFN-γ, IL-4, and IL-17 were utilized to detect the frequencies of T H 1 ( a ), T H 2 ( b ), and T H 17 ( c ) cells among CD4+ T-cell population, respectively, by flow cytometry analysis. While T H 1 percentages were reduced, T H 2 and T H 17 cell ratios stayed unaltered in NK cell-depleted (NK−) mice in comparison to the control mice (NK+). ** represents p < 0.01.

Article Snippet: Of those cells, 5 × 10 6 were removed and added into 15 mL falcon tubes for flow cytometry analysis of CD4+ T-cell subset levels after 24-h treatment with 5 μg/mL PPV-23 vaccine antigen and addition of 5 μg/mL brefeldin A (BioLegend, 420601) at 16th hour of incubation.

Techniques: Comparison, Flow Cytometry

Journal: International Archives of Allergy and Immunology

Article Title: Importance of NK Cells in Cellular and Humoral Responses Triggered by Pneumococcus Vaccination

doi: 10.1159/000535562

Figure Lengend Snippet: Gating strategy to identify T H populations of interest. Lymphocyte populations were gated based on FSC/SSC, excluding aggregates. T H cells were then gated from CD3+CD4+ cells, and T H 1, T H 2, and T H 17 cell populations were identified based on IFN-γ, IL-4, and IL-17 positivity, respectively.

Article Snippet: Splenocytes were then incubated with fluorochrome-conjugated monoclonal antibodies against CD3 (Cytek Biosciences; for total T-cells), CD4 (Cytek Biosciences; for T H cells), IFN-γ (Biolegend; for T H 1 cells), IL-4 (Biolegend; for T H 2 cells), and IL-17 (Biolegend; for T H 17 cells) for 20 min in the dark at room temperature.

Techniques:

Journal: International Archives of Allergy and Immunology

Article Title: Importance of NK Cells in Cellular and Humoral Responses Triggered by Pneumococcus Vaccination

doi: 10.1159/000535562

Figure Lengend Snippet: Comparison of CD4+ T-cell subset levels. Splenocytes collected 4 weeks after the PPV-23 vaccination were stimulated by PPV-23 vaccine antigen for 24 h. Antibodies against IFN-γ, IL-4, and IL-17 were utilized to detect the frequencies of T H 1 ( a ), T H 2 ( b ), and T H 17 ( c ) cells among CD4+ T-cell population, respectively, by flow cytometry analysis. While T H 1 percentages were reduced, T H 2 and T H 17 cell ratios stayed unaltered in NK cell-depleted (NK−) mice in comparison to the control mice (NK+). ** represents p < 0.01.

Article Snippet: Splenocytes were then incubated with fluorochrome-conjugated monoclonal antibodies against CD3 (Cytek Biosciences; for total T-cells), CD4 (Cytek Biosciences; for T H cells), IFN-γ (Biolegend; for T H 1 cells), IL-4 (Biolegend; for T H 2 cells), and IL-17 (Biolegend; for T H 17 cells) for 20 min in the dark at room temperature.

Techniques: Comparison, Flow Cytometry