apc conjugated mouse anti chicken cd3  (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: APC-conjugated mouse anti-chicken CD3+, PE-conjugated mouse anti-chicken CD8α+, FITC-conjugated mouse anti-chicken CD4+, (SouthernBiotech, Birmingham, USA) were used in this study.

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

Journal: The Journal of Biological Chemistry

Article Title: Revealing novel and conservative T-cell epitopes with MHC B2 restriction on H9N2 avian influenza virus (AIV)

doi: 10.1016/j.jbc.2024.107395

Figure Lengend Snippet: Detection of immune response in B2 haplotype chickens after H9N2 AIV infected. A , gating strategy of T lymphocyte from PBMCs after H9N2 AIV infection. B – D , four chickens from the infected and control groups were randomly selected for sampling to detect the percentage of CD3 + CD8α + T cells ( B ), the percentage of CD3 + CD4 + T cells ( C ), and the ratio of CD3 + CD4 + CD8α + T cells ( D ). 1.5 × 10 5 cells per sample were collected for flow cytometric analysis. Expressions of immune-related genes in PBMCs were detected by quantitative real-time polymerase chain reaction (qRT-PCR). In the H9N2 AIV-infected experiments, the total RNA of PBMC was extracted from chickens at 5 DPI. E – G , the transcriptional expressions of immune-related genes, including cytotoxicity-associated genes ( E ), innate immunity-related genes ( F ), and Th2 genes ( G ) were analyzed. Furthermore, data were collected from three biological samples in each group, and each sample performed in triplicate. The results were presented as means ± SEM, and the unpaired t test was used for statistical comparison of T lymphocyte percentage, and the paired t test was used for statistical comparison of expressions of immune-related genes. ∗p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. AIV, avian influenza virus; DPI, days postinfection; IFN-γ, interferon-γ; PBMCs, peripheral blood mononuclear cells.

Article Snippet: 1 × 10 6 PBMCs were incubated with allophycocyanin-conjugated mouse anti-chicken CD3 + , FITC-conjugated mouse anti-chicken CD4 + , and PE-conjugated mouse anti-chicken CD8α + monoclonal antibodies (SouthernBiotech) in the dark at 4 °C for 30 min. After three washes with PBS, the ratios of CD3 + CD4 + T cells, CD3 + CD8α + T cells, and CD3 + CD4 + CD8α + T cells were analyzed by flow cytometry (CytoFLEX, Beckman Coulter).

Techniques: Infection, Control, Sampling, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Comparison, Virus