mouse anti chicken cd4 mab  (SouthernBiotech)

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: Frontiers in Immunology

Article Title: Marek’s Disease Virus Modulates T Cell Proliferation via Activation of Cyclooxygenase 2-Dependent Prostaglandin E2

doi: 10.3389/fimmu.2021.801781

Figure Lengend Snippet: MDV infected CEFs and 265L tumour cells produce high PGE2 level resulting in downregulation of chIL2 and transferrin uptake in CD3+ T cells. Culture supernatants were harvested from control CEF (non-infected),CEF infected with virulent MDV (MDV supernatant) or vaccine strain of MDV (vaccine supernatant) at 72 hrs post-infection. Culture supernatant harvested from CD4+ T cells of naïve splenocytes and MDV-transformed CD4+ T cells (265L supernatant). (A) The levels of PGE2 were determined using an ELISA assay. (B) chIL2 expression level was determined in Con-A stimulated splenocytes treated with PGE2 (10 µg/ml), or the supernatants as described above using qRT-PCR, and fold change are shown. Inhibition of chIL-2 expression by (C) PGE2, (D) MDV supernatant, (E) 265L supernatant was rescued in the presence of the inhibitors of the COX-2/PGE2 pathway; TG4-155 (4 μM), ER-819762 (8 μM) and SC-236 (5 μg/mL). (F) Representative confocal images showing uptake of transferrin (red color) by CD3+ T cells in the Con-A stimulated splenocytes treated with control supernatant, PGE2, MDV supernatant, vaccine supernatant or 265L supernatant using confocal microscopy. (G) Graph shows mean fluorescent intensity (MFI) of transferrin uptake using confocal microscopy. (H) Bar graph shows the effects of a COX-2 inhibitor (SC-236) on transferrin uptake in CD3+ T cells treated with PGE2, MDV supernatant or 265L supernatant using confocal microscopy. Graphs are representative of three independent experiments, each performed with three biological replicates. Grey bars are used in the experimental groups in which no significant difference are found, while red bars are used to represent significant differences. Each dot in ELISA represents the average of three biological replicates from six individual culture groups. Error bar represents the mean ± standard deviation. Statistical significance was estimated as p value calculated by ANOVA test. NS, not significant.

Article Snippet: Briefly, chicken splenocytes (1 × 10 7 ) were incubated (30 min at 4°C) with PE-conjugated mouse anti-chicken CD3 mAbs or PE-conjugated mouse anti-chicken CD4 mAb (10 μl/10 7 cells) (Cambridge Bioscience, Cambridge, UK).

Techniques: Infection, Transformation Assay, Enzyme-linked Immunosorbent Assay, Expressing, Quantitative RT-PCR, Inhibition, Confocal Microscopy, Standard Deviation

Journal: Veterinary World

Article Title: Development of lymphocyte subpopulations in local breed chickens

doi: 10.14202/vetworld.2021.1846-1852

Figure Lengend Snippet: Thymus and bursa of Fabricius sections showing (a) isotype control. (b) Bursa of Fabricius cell showing lymphocytes expressing Bu-1 marker (brown in color). (c) Thymocytes expressing CD4 marker (brown-stained cells). (d) Thymocytes expressing CD8 marker (brown-stained cells). Primary antibody and biotinylated horse anti-mouse IgG secondary antibody were the first to treat the section. A ”charged” DAB (Abcam, USA) was the primary stain to develop color and methyl green was the counterstain.

Article Snippet: Alternatively, mouse anti-chicken CD4-FITC-conjugated mAb (mouse IgG1) (Southern Biotech) and mouse anti-chicken CD8-PE-conjugated mAb (mouse IgG1) (Southern Biotech) were used in the two-color, direct immunofluorescent staining procedure to identify the CD4 + and/or CD8 + markers, respectively, on the T lymphocytes.

Techniques: Control, Expressing, Marker, Staining

Journal: Veterinary World

Article Title: Development of lymphocyte subpopulations in local breed chickens

doi: 10.14202/vetworld.2021.1846-1852

Figure Lengend Snippet: Spleen section showing: (a) Isotype control. (b) CD8 (CD4 – CD8 + ) lymphocyte stained in brown color especially in red bulb (RP) 40×. (c) Lymphocytes expressing CD3 marker 100×. (d) Lymphocytes expressing Bu-1 marker 100×. Primary antibody and biotinylated horse anti-mouse IgG secondary antibody were the first to treat the section. A ”charged” DAB (Abcam, USA) was the primary stain to develop color and methyl green was the counterstain.

Article Snippet: Alternatively, mouse anti-chicken CD4-FITC-conjugated mAb (mouse IgG1) (Southern Biotech) and mouse anti-chicken CD8-PE-conjugated mAb (mouse IgG1) (Southern Biotech) were used in the two-color, direct immunofluorescent staining procedure to identify the CD4 + and/or CD8 + markers, respectively, on the T lymphocytes.

Techniques: Control, Staining, Expressing, Marker

Journal: Veterinary World

Article Title: Development of lymphocyte subpopulations in local breed chickens

doi: 10.14202/vetworld.2021.1846-1852

Figure Lengend Snippet: Flow cytometry results of cell suspension from spleen, thymus, and bursa of Fabricius. T lymphocytes (CD8 in particular with CD4 – CD8 + marker) were the dominant cells in the spleen. Thymocytes show that the CD3 marker is denser than that in bursa. Conversely, the Bu-1 marker is highly expressed by the lymphocytes in bursa. All the lymphocytes in the thymus were expressing both CD4 and CD8 markers (CD4 + CD8 + ) with traces of CD4 – CD8 - , CD4 + CD8 – , and CD4 – CD8 + . In the bursa, almost all the lymphocytes are expressing no marker (CD4 – CD8 + ).

Article Snippet: Alternatively, mouse anti-chicken CD4-FITC-conjugated mAb (mouse IgG1) (Southern Biotech) and mouse anti-chicken CD8-PE-conjugated mAb (mouse IgG1) (Southern Biotech) were used in the two-color, direct immunofluorescent staining procedure to identify the CD4 + and/or CD8 + markers, respectively, on the T lymphocytes.

Techniques: Flow Cytometry, Suspension, Marker, Expressing

Journal: Veterinary World

Article Title: Development of lymphocyte subpopulations in local breed chickens

doi: 10.14202/vetworld.2021.1846-1852

Figure Lengend Snippet: Determination of lymphocyte population subsets in lymphoid organs.

Article Snippet: Alternatively, mouse anti-chicken CD4-FITC-conjugated mAb (mouse IgG1) (Southern Biotech) and mouse anti-chicken CD8-PE-conjugated mAb (mouse IgG1) (Southern Biotech) were used in the two-color, direct immunofluorescent staining procedure to identify the CD4 + and/or CD8 + markers, respectively, on the T lymphocytes.

Techniques: