Journal: Frontiers in Immunology

Article Title: Generation and characterization of chicken monocyte-derived dendritic cells

doi: 10.3389/fimmu.2025.1517697

Figure Lengend Snippet: List of antibodies.

Article Snippet: Mouse anti-chicken MHCII (Clone 2G11) , IgG1 , FITC , 8350-02 , Southern Biotech , 10 μg/mL.

Techniques: Control

Journal: Frontiers in Immunology

Article Title: Generation and characterization of chicken monocyte-derived dendritic cells

doi: 10.3389/fimmu.2025.1517697

Figure Lengend Snippet: Phenotype analysis of chicken adherent PBMCs cultured 5 days in presence of GM-CSF and IL-4. (A) Gating strategy for flow cytometry analysis of chicken MoDCs based on FSC and SSC properties and singlets were selected from the FSC-A versus FSC-H. Dead cells were excluded using a viability dye. (B) Representative histograms of putative CD11c expression in adherent PBMCs incubated for 5 days only with non-supplemented complete medium (white) or in the presence of GM-CSF and IL-4 (blue). The background staining was evaluated using an isotype control (grey). Numbers on histograms represent the percentage of putative CD11c+ among live cells. (C) Boxplot of the percentage of putative CD11c+ cells among viable cells. Each symbol (circle or square) represents an individual chicken in the corresponding condition (n = 8). **p < 0.005, two-tailed paired t-test. (D) Representative contour plots of MHCII expression in adherent PBMCs incubated for 5 days with non-supplemented complete medium or in the presence of GM-CSF and IL-4. Numbers represent the percentage of MHCII+ cells among viable cells. (E) Boxplot of the percentage of MHCII+ cells among viable cells. Each symbol (circle or square) represents an individual chicken in the corresponding condition (n=8). A non-parametric Wilcoxon matched-pairs signed rank test was used to determine statistical differences (**p < 0.005).

Article Snippet: Mouse anti-chicken MHCII (Clone 2G11) , IgG1 , FITC , 8350-02 , Southern Biotech , 10 μg/mL.

Techniques: Cell Culture, Flow Cytometry, Expressing, Incubation, Staining, Control, Two Tailed Test

Journal: Frontiers in Immunology

Article Title: Generation and characterization of chicken monocyte-derived dendritic cells

doi: 10.3389/fimmu.2025.1517697

Figure Lengend Snippet: LPS stimulation increased MHCII and co-stimulatory molecules expressions, and IL-12p40 secretion. Flow cytometry was used to analyze unstimulated and LPS-stimulated MoDCs for their expression of MHCII, CD80, and CD40 after 6 and 24 h (A) Representative contour plots of MHCII with numbers on plots representing the percentage of MHCII+ cells gated on live cells and the MFI of MHCII+ cells. (B) The upper panel represents the percentage of MHCII+ cells among viable cells. Each circle represents an individual chicken in the corresponding condition. A non-parametric Wilcoxon matched-pairs signed rank test was used to determine statistical differences (ns, not significant). The lower panel represents MHCII MFI determined on viable MHCII+ cells. Each circle represents an individual chicken in the corresponding condition. A two-tailed paired t-test was used to determine statistical differences (****p<0.0001). (C) Representative flow cytometry histograms of CD80 (left panel), and CD40 (right panel) expressions in unstimulated cells (blue), LPS-stimulated MoDCs (orange), and isotype control (grey) after 6 and 24 h Numbers on histograms indicate the MFI values of the corresponding marker. (D) Boxplot of CD80 (upper panel) and CD40 (bottom panel) expression measured as MFI normalized to the isotype control. Each circle represents an individual chicken in the corresponding condition. A two-tailed paired t-test and a non-parametric Wilcoxon matched-pairs signed rank test were used to determine statistical differences (**p<0.01; ***p<0.001). The data from 3 independent experiments are displayed (n=14). (E) The production of IL-12p40 was quantified in the supernatants of unstimulated or LPS-stimulated MoDCs at 6 and 24 h by ELISA. Data are presented as boxplots with individual chicken values represented by circles. A two-tailed paired t-test was used to determine statistical differences (***p<0.001; ****p<0.0001). Data from two independent experiments (n=8) are displayed.

Article Snippet: Mouse anti-chicken MHCII (Clone 2G11) , IgG1 , FITC , 8350-02 , Southern Biotech , 10 μg/mL.

Techniques: Flow Cytometry, Expressing, Two Tailed Test, Control, Marker, Enzyme-linked Immunosorbent Assay

Journal: Veterinary research

Article Title: Revealing novel and conservative CD8 + T-cell epitopes with MHC B2 restriction on ALV-J.

doi: 10.1186/s13567-024-01426-3

Figure Lengend Snippet: Figure 1 Detection of ALV-J shedding, ALV-J viremia, ALV-J antibody and T lymphocyte percentage of B2 haplotype chickens after ALV-J infection. Six chickens were randomly selected for sampling every 7 days post-infection (dpi). A ALV-J shedding was monitored via detecting the p27 expression levels in cloacal swabs. S/P value below 0.2 indicated negative ALV-J shedding. B ALV-J viremia was monitored via virus isolation. An S/P value above 0.2 indicated positive ALV-J viremia. C The ALV-J antibody level in the serum was monitored using the commercial ALV-J antibody test kit. An S/P value above 0.6 was considered ALV-J antibody positive. Five days before infection (dbi) and each week after infection, PBMCs derived from five chickens of infected and control groups were isolated to detect the T lymphocyte percentage, including the percentage of the CD4+CD3+T cell (D), the CD8α+CD3+T cell (E), and the CD3+CD4+CD8α+T cell (F). Each sample collected 1 × 105 cells for flow cytometric analysis. The one-way test was used for statistical comparison among A, B and C. And the unpaired Student t test was used for statistical comparison among D, E and F. * P < 0.05, ** P < 0.01.

Article Snippet: PBMCs (1 × 106) were incubated with mouse anti-chicken CD3 mAb (SouthernBiotech, Birmingham, USA), mouse anti-chicken CD4 mAb (SouthernBiotech, Birmingham, USA) and mouse anti-chicken CD8α mAb (SouthernBiotech, Birmingham, USA), respectively, at 4 °C for 30 min in the dark.

Techniques: Infection, Sampling, Expressing, Virus, Isolation, Derivative Assay, Control, Comparison

Journal: Veterinary research

Article Title: Revealing novel and conservative CD8 + T-cell epitopes with MHC B2 restriction on ALV-J.

doi: 10.1186/s13567-024-01426-3

Figure Lengend Snippet: Figure 1 Detection of ALV-J shedding, ALV-J viremia, ALV-J antibody and T lymphocyte percentage of B2 haplotype chickens after ALV-J infection. Six chickens were randomly selected for sampling every 7 days post-infection (dpi). A ALV-J shedding was monitored via detecting the p27 expression levels in cloacal swabs. S/P value below 0.2 indicated negative ALV-J shedding. B ALV-J viremia was monitored via virus isolation. An S/P value above 0.2 indicated positive ALV-J viremia. C The ALV-J antibody level in the serum was monitored using the commercial ALV-J antibody test kit. An S/P value above 0.6 was considered ALV-J antibody positive. Five days before infection (dbi) and each week after infection, PBMCs derived from five chickens of infected and control groups were isolated to detect the T lymphocyte percentage, including the percentage of the CD4+CD3+T cell (D), the CD8α+CD3+T cell (E), and the CD3+CD4+CD8α+T cell (F). Each sample collected 1 × 105 cells for flow cytometric analysis. The one-way test was used for statistical comparison among A, B and C. And the unpaired Student t test was used for statistical comparison among D, E and F. * P < 0.05, ** P < 0.01.

Article Snippet: PBMCs (1 × 106) were incubated with mouse anti-chicken CD3 mAb (SouthernBiotech, Birmingham, USA), mouse anti-chicken CD4 mAb (SouthernBiotech, Birmingham, USA) and mouse anti-chicken CD8α mAb (SouthernBiotech, Birmingham, USA), respectively, at 4 °C for 30 min in the dark.

Techniques: Infection, Sampling, Expressing, Virus, Isolation, Derivative Assay, Control, Comparison

Journal: Veterinary research

Article Title: Revealing novel and conservative CD8 + T-cell epitopes with MHC B2 restriction on ALV-J.

doi: 10.1186/s13567-024-01426-3

Figure Lengend Snippet: Figure 1 Detection of ALV-J shedding, ALV-J viremia, ALV-J antibody and T lymphocyte percentage of B2 haplotype chickens after ALV-J infection. Six chickens were randomly selected for sampling every 7 days post-infection (dpi). A ALV-J shedding was monitored via detecting the p27 expression levels in cloacal swabs. S/P value below 0.2 indicated negative ALV-J shedding. B ALV-J viremia was monitored via virus isolation. An S/P value above 0.2 indicated positive ALV-J viremia. C The ALV-J antibody level in the serum was monitored using the commercial ALV-J antibody test kit. An S/P value above 0.6 was considered ALV-J antibody positive. Five days before infection (dbi) and each week after infection, PBMCs derived from five chickens of infected and control groups were isolated to detect the T lymphocyte percentage, including the percentage of the CD4+CD3+T cell (D), the CD8α+CD3+T cell (E), and the CD3+CD4+CD8α+T cell (F). Each sample collected 1 × 105 cells for flow cytometric analysis. The one-way test was used for statistical comparison among A, B and C. And the unpaired Student t test was used for statistical comparison among D, E and F. * P < 0.05, ** P < 0.01.

Article Snippet: PBMCs (1 × 106) were incubated with mouse anti-chicken CD3 mAb (SouthernBiotech, Birmingham, USA), mouse anti-chicken CD4 mAb (SouthernBiotech, Birmingham, USA) and mouse anti-chicken CD8α mAb (SouthernBiotech, Birmingham, USA), respectively, at 4 °C for 30 min in the dark.

Techniques: Infection, Sampling, Expressing, Virus, Isolation, Derivative Assay, Control, Comparison