pe conjugated mouse anti chicken cd8α  (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: 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: Journal of Advanced Veterinary and Animal Research

Article Title: Humoral, cellular immunity and efficacy of bioreactor propagated and inactivated Fowl adenovirus 8b adjuvanted with Montanide 71VG in broiler chickens

doi: 10.5455/javar.2024.k819

Figure Lengend Snippet: Dot-plots of the flow cytometric analysis of CD3+, CD4+, and CD8+ sub-population in the liver, spleen, and thymus of challenged chickens inoculated with inactivated FAdV 8b UPM11142P5B1 at 35 dpi.

Article Snippet: After being aliquoted to a Falcon tube (FALCON-Corning, NC, USA), the cells corresponding to 1×10 6 /ml from each sample were stained with mouse anti-chicken CD3-FITC, mouse anti-chicken CD4-APC, and mouse anti-chicken CD8α-PE antibodies (SouthernBiotech, Birmingham, AL, USA).

Techniques:

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

Journal: Poultry Science

Article Title: Inhibitory effect of Eimeria maxima IFN-γ inhibitory molecules on the immune function of T cell subsets in chickens

doi: 10.1016/j.psj.2023.103098

Figure Lengend Snippet: Detection of the sorting purity of chicken CD8 + , CD4 + , CD4 + CD25 − and CD4 + CD25 + T cells by flow cytometry. (A) Magnetic bead sorting of chicken CD8 + T cells A1: blank cells (total PBMC); A2: CD8 + T cells before cell sorting; A3: CD8 + T cells after cell sorting. (B) Magnetic bead sorting of chicken CD4 + T cells B1: blank cells (total PBMC); B2: CD4 + T cells before cell sorting; B3: CD4 + T cells after cell sorting. (C) Magnetic bead sorting of chicken CD4 + CD25 − T cells C1: blank cells (total PBMC); C2: CD4 + CD25 − T cells before cell sorting; C3: CD4 + CD25 − T cells after cell sorting. (D) Magnetic bead sorting of chicken CD4 + CD25 + T cells D1: blank cells (total PBMC); D2: CD25 + cells before cell sorting D3: CD25 + cells after cell sorting D4: CD4 + CD25 + T cells before cell sorting; D5: CD4 + CD25 + T cells after cell sorting.

Article Snippet: The primary phycoerythrin ( PE )-conjugated mouse anti-chicken CD8α or CD4 antibody (Southernbiotech, Birmingham, AL) was added according to 1 × 10 8 cells.

Techniques: Flow Cytometry, FACS

Journal: Poultry Science

Article Title: Inhibitory effect of Eimeria maxima IFN-γ inhibitory molecules on the immune function of T cell subsets in chickens

doi: 10.1016/j.psj.2023.103098

Figure Lengend Snippet: Effects of rEmHPSP-2 and rEmHPSP-3 on the transcription of IFN-γ and IL-2 in chicken CD8 + T cells (A) Effects of rEmHPSP-2 on the transcription of IFN-γ and IL-2 in chicken CD8 + T cells. (B) Effects of rEmHPSP-3 on the transcription of IFN-γ and IL-2 in chicken CD8 + T cells. The differences between groups were analyzed using Duncan method of 1-way ANOVA, where the same letter indicates no significant difference ( P > 0.05) and different letter indicates significant difference ( P < 0.05).

Article Snippet: The primary phycoerythrin ( PE )-conjugated mouse anti-chicken CD8α or CD4 antibody (Southernbiotech, Birmingham, AL) was added according to 1 × 10 8 cells.

Techniques:

Journal: Vaccines

Article Title: Construction and Evaluation of the Immunogenicity and Protective Efficacy of Recombinant Replication-Deficient Human Adenovirus-5 Expressing Genotype VII Newcastle Disease Virus F Protein and Infectious Bursal Disease Virus VP2 Protein

doi: 10.3390/vaccines11061051

Figure Lengend Snippet: Comparison of cellular immunity in different groups: ( A ) Statistical analysis of the peripheral blood lymphocyte stimulation index in the ND groups. Under ConA stimulation, the SI values of peripheral blood lymphocytes in the PBS group were significantly lower than those of the rDHN3-mF, rAd5-F, and rAd5-VP2-F2A-F groups ( p < 0.001). The SI values of peripheral blood lymphocytes in the rAd5-EGFP group were significantly lower than those in the rDHN3-mF, rAd5-F, and rAd5-VP2-F2A-F groups ( p < 0.01). However, there was no significant difference between the rDHN3-mF, rAd5-F, and rAd5-VP2-F2A-F groups ( p > 0.05), and no significant differences between the PBS and rAd5-EGFP groups ( p > 0.05). Under inactivated NDV stimulation, the SI values of peripheral blood lymphocytes in the PBS and rAd5-EGFP groups were significantly lower than those in the rDHN3-mF, rAd5-F, and rAd5-VP2-F2A-F groups ( p < 0.0001). SI values in the rAd-VP2-F2A-F group were significantly different from those of the rDHN3-mF and rAd5-F groups ( p < 0.05); no significant differences between the rDHN3-mF and rAd5-F groups ( p > 0.05), and there was no significant difference between the PBS and rAd5-EGFP groups ( p > 0.05) (ns—non-significant; * p < 0.05); ( B ) Statistical analysis of the peripheral blood lymphocyte stimulation index in IBD groups. From ( D ) above, it can be seen that the SI values of peripheral blood lymphocytes in the HVT-VP2 vector vaccines group, rAd5-VP2 group, and rAd5-VP2-F2A-F group were not significantly different from each other regardless of ConA stimulation or inactivated IBDV stimulation in the IBD group ( p > 0.05). However, the SI values of the above three groups were significantly higher than those of the rAd5- EGFP and the PBS groups ( p < 0.05). There was no significant difference between the PBS and rAd5-EGFP groups ( p > 0.05) (ns—non-significant); ( C , D ) Statistical analysis of the percentage of CD4+ and CD8+ T lymphocytes in the peripheral blood in ND groups. It can be seen from these two pictures that the percentages of CD4+ and CD8+ T lymphocytes in the rAd5-F, rDHN3-mF, and rAd5-VP2-F2A-F groups were significantly higher than those in the PBS group ( p < 0.05). There was no significant difference between the above three vaccine groups ( p > 0.05); ( E , F ) Statistical analysis of the percentage of CD4+ and CD8+ T lymphocytes in the peripheral blood in the IBD groups. It can be seen from these two pictures that the percentage of CD4+ and CD8+ T lymphocytes in the rAd5-VP2 group, the HVT-VP2 vector vaccines, and rAd5-VP2-F2A-F groups were significantly higher than in the PBS group ( p < 0.05). There was no significant difference between the three vaccine groups ( p > 0.05).

Article Snippet: The T-lymphocyte subpopulations were analyzed by flow cytometry (Beckman Coulter, Carlsbad, CA, USA) with the following antibodies: mouse anti-chicken CD3, mouse anti-chicken CD4, and mouse anti-chicken CD8α (Southern Biotech, Birmingham, AL, USA).

Techniques: Comparison, Plasmid Preparation, Vaccines