Journal: Viruses

Article Title: Role of T Cells in Vaccine-Mediated Immunity against Marek's Disease.

doi: 10.3390/v15030648

Figure Lengend Snippet: Figure 1. T Cells depletion. Flow cytograms show the percentage of CD4+ and CD8+ T Cells in the control (Panel A), CD4+ T Cell depleted birds (Panel B), CD8+ T Cell depleted birds (Panel C), and CD4+/CD8+ T Cell depleted birds (Panel D) 11 days post-treatment. Blood samples from three birds per group were pooled, PBMC isolated, and 1 × 106 cells/100 µL was used for cell surface antigen analysis. The CD4+ T Cells were stained with CD4-PE, and CD8+ T Cells were stained with CD8α-FITC,

Article Snippet: Anti-CD4 mononuclear cell binding specificity. (A): Histopaque 1077-treated PBMC (1 × 106 cells) with no added antibodies (negative control); (B): PBMC stained with mouse anti-chicken CD4-PE antibody (Southern Biotech, positive control); (C): PBMC stained with rat anti-mouse IgMPE/CY7 antibody (secondary antibody only); (D): PBMC stained with primary monoclonal antibody isolated from hybridoma cell line (IgM, at 1.5 μg per 1 × 106 cells) and the secondary rat anti-mouse IgM-PE/CY7 antibody; (E): PBMC stained with primary monoclonal antibody isolated from hybridoma cell line (IgM, at 0.298 μg per 1 × 106) and the secondary rat anti-mouse IgM-PE/CY7 antibody.

Techniques: Control, Isolation, Staining

Journal: Viruses

Article Title: Role of T Cells in Vaccine-Mediated Immunity against Marek's Disease.

doi: 10.3390/v15030648

Figure Lengend Snippet: Figure 2. Recovery of CD4+ and CD8+ T Cells 13 days post-termination of antibody treatment. The percentage population of CD4+ and CD8+ T Cells in the control birds (Panel A), CD4+ T Cell depleted birds (Panel B), and CD8+ T Cell depleted group (Panel C) are depicted 13 days post-termination of antibody treatment. Blood samples from three birds per group were pooled, PBMC isolated, and 1 × 106 cells/100 µL was used for cell surface antigen analysis. The CD4+ T Cells were stained with CD4-PE, and CD8+ T Cells were stained with CD8α -FITC, 11–39 monoclonal antibodies. (Panel D) Bar graphs showing the percentage of B and T Cell populations 13 days after termination of antibody treatment. Comparative analysis was made between the untreated control and the T Cell depleted birds. Same total blood samples were used for the staining of B cells and double staining of CD4+, and CD8+ T Cells. B cells, CD4+ T Cells, and CD8+ T Cells were stained with monoclonal antibodies Bu1-RPE, CD4-PE, and CD8α -FITC, respectively. V: vaccinated; C: challenged.

Article Snippet: Anti-CD4 mononuclear cell binding specificity. (A): Histopaque 1077-treated PBMC (1 × 106 cells) with no added antibodies (negative control); (B): PBMC stained with mouse anti-chicken CD4-PE antibody (Southern Biotech, positive control); (C): PBMC stained with rat anti-mouse IgMPE/CY7 antibody (secondary antibody only); (D): PBMC stained with primary monoclonal antibody isolated from hybridoma cell line (IgM, at 1.5 μg per 1 × 106 cells) and the secondary rat anti-mouse IgM-PE/CY7 antibody; (E): PBMC stained with primary monoclonal antibody isolated from hybridoma cell line (IgM, at 0.298 μg per 1 × 106) and the secondary rat anti-mouse IgM-PE/CY7 antibody.

Techniques: Control, Isolation, Staining, Bioprocessing, Double Staining

Journal: Viruses

Article Title: Role of T Cells in Vaccine-Mediated Immunity against Marek's Disease.

doi: 10.3390/v15030648

Figure Lengend Snippet: Figure 3. PCR-based analysis of viral DNA in spleen samples of control and treated birds at 5 days post-inoculation (dpi, Panel A), 10 dpi (Panel B), 20 dpi (Panel C), and 57 dpi (Panel D). The viral genome detection in the non-vaccinated challenged birds (Lanes 14, 15, and 16) is depicted by green arrows. The detection of pp38 in the T Cell depleted, vaccinated, and challenged birds (lanes 2–13) is shown by red arrows. Lanes: M, DNA ladder, 1: Control bird, 2–4: Birds with intact T Cell, vaccinated, challenged, 5–7: Birds with CD4+ T Cell depleted, vaccinated, challenged, 8–10: Birds with CD8+ T Cell depleted, vaccinated, challenged, 11–13: Birds with CD4+/CD8+ T Cell depleted, vaccinated, challenged, 14–16: Birds with intact T Cells, non-vaccinated, challenged, 17: Positive control for pp38 amplification using MDV DNA isolated from infected birds (blue arrow), 18: GAPDH (blue arrow), M: DNA ladder.

Article Snippet: Anti-CD4 mononuclear cell binding specificity. (A): Histopaque 1077-treated PBMC (1 × 106 cells) with no added antibodies (negative control); (B): PBMC stained with mouse anti-chicken CD4-PE antibody (Southern Biotech, positive control); (C): PBMC stained with rat anti-mouse IgMPE/CY7 antibody (secondary antibody only); (D): PBMC stained with primary monoclonal antibody isolated from hybridoma cell line (IgM, at 1.5 μg per 1 × 106 cells) and the secondary rat anti-mouse IgM-PE/CY7 antibody; (E): PBMC stained with primary monoclonal antibody isolated from hybridoma cell line (IgM, at 0.298 μg per 1 × 106) and the secondary rat anti-mouse IgM-PE/CY7 antibody.

Techniques: Control, Positive Control, Isolation, Infection

Journal: Viruses

Article Title: Role of T Cells in Vaccine-Mediated Immunity against Marek's Disease.

doi: 10.3390/v15030648

Figure Lengend Snippet: Figure 4. Anti-CD4 mononuclear cell binding specificity. (A): Histopaque 1077-treated PBMC (1 × 106 cells) with no added antibodies (negative control); (B): PBMC stained with mouse anti- chicken CD4-PE antibody (Southern Biotech, positive control); (C): PBMC stained with rat anti-mouse IgM-PE/CY7 antibody (secondary antibody only); (D): PBMC stained with primary monoclonal antibody isolated from hybridoma cell line (IgM, at 1.5 µg per 1 × 106 cells) and the secondary rat anti-mouse IgM-PE/CY7 antibody; (E): PBMC stained with primary monoclonal antibody isolated from hybridoma cell line (IgM, at 0.298 µg per 1 × 106) and the secondary rat anti-mouse IgM-PE/CY7 antibody. The gated green cells in the middle of panels (D,E) are staining the same population of cells as in the middle of panel B.

Article Snippet: Anti-CD4 mononuclear cell binding specificity. (A): Histopaque 1077-treated PBMC (1 × 106 cells) with no added antibodies (negative control); (B): PBMC stained with mouse anti-chicken CD4-PE antibody (Southern Biotech, positive control); (C): PBMC stained with rat anti-mouse IgMPE/CY7 antibody (secondary antibody only); (D): PBMC stained with primary monoclonal antibody isolated from hybridoma cell line (IgM, at 1.5 μg per 1 × 106 cells) and the secondary rat anti-mouse IgM-PE/CY7 antibody; (E): PBMC stained with primary monoclonal antibody isolated from hybridoma cell line (IgM, at 0.298 μg per 1 × 106) and the secondary rat anti-mouse IgM-PE/CY7 antibody.

Techniques: Binding Assay, Negative Control, Staining, Positive Control, Isolation

Journal: Viruses

Article Title: Role of T Cells in Vaccine-Mediated Immunity against Marek's Disease.

doi: 10.3390/v15030648

Figure Lengend Snippet: Figure 6. Immunohistochemical analysis of MDV antigen in the skin samples of all vaccinated and challenged groups with intact or depleted T Cells. Anti-gB monoclonal antibody was used for detection of virus particles in the skin tissues of challenged groups. (Panel A) depicts skin sample from an unvaccinated, challenged bird with intact T Cells showing significant viral replication in the FFE (blue arrow). (Panel B) represents the skin sample from a vaccinated/challenged bird with intact T Cells showing minor MDV antigen in the FFE (arrows). (Panel C) depicts skin sample from a CD4+ T Cell depleted, vaccinated/challenged bird that exhibits minor viral replication in the FFE (blue arrow). The replication rate of MDV in the skin of a CD8+ T Cell depleted bird is depicted in (Panel D) (arrows). (Panel E) shows the replication rate of MDV in the skin sample of a CD4+/CD8+

Article Snippet: Anti-CD4 mononuclear cell binding specificity. (A): Histopaque 1077-treated PBMC (1 × 106 cells) with no added antibodies (negative control); (B): PBMC stained with mouse anti-chicken CD4-PE antibody (Southern Biotech, positive control); (C): PBMC stained with rat anti-mouse IgMPE/CY7 antibody (secondary antibody only); (D): PBMC stained with primary monoclonal antibody isolated from hybridoma cell line (IgM, at 1.5 μg per 1 × 106 cells) and the secondary rat anti-mouse IgM-PE/CY7 antibody; (E): PBMC stained with primary monoclonal antibody isolated from hybridoma cell line (IgM, at 0.298 μg per 1 × 106) and the secondary rat anti-mouse IgM-PE/CY7 antibody.

Techniques: Immunohistochemical staining, Virus

Journal: Viruses

Article Title: Role of T Cells in Vaccine-Mediated Immunity against Marek's Disease.

doi: 10.3390/v15030648

Figure Lengend Snippet: Figure 7. The picture depicts the chest bone (keeled sternum) of a CD4+/CD8+ T Cell depleted bird that is severely emaciated (Panel A). These birds exhibit no clinical signs of MD during the experiment and no T Cell lymphoma at termination. The birds experienced breathing difficulties. (Panel B) shows the spleen of a CD4+/CD8+ T Cell depleted bird at termination. Left: spleen from a CD4+ T Cell depleted bird; right: spleen from CD4+/CD8+ T Cell depleted bird. This contrasts with MDV-infected birds where the spleen is enlarged (splenomegaly), and the thymus and bursa are atrophied. (Panel C) depicts the bursa of a CD4+/CD8+ T Cell depleted bird. Although the spleen tissues from these birds were negative for MDV genome, the bursas, like the spleens, were severely atrophied. Left: bursa from a CD4+ T Cell depleted bird; right: bursa from a CD4+/CD8+ T Cell depleted bird.

Article Snippet: Anti-CD4 mononuclear cell binding specificity. (A): Histopaque 1077-treated PBMC (1 × 106 cells) with no added antibodies (negative control); (B): PBMC stained with mouse anti-chicken CD4-PE antibody (Southern Biotech, positive control); (C): PBMC stained with rat anti-mouse IgMPE/CY7 antibody (secondary antibody only); (D): PBMC stained with primary monoclonal antibody isolated from hybridoma cell line (IgM, at 1.5 μg per 1 × 106 cells) and the secondary rat anti-mouse IgM-PE/CY7 antibody; (E): PBMC stained with primary monoclonal antibody isolated from hybridoma cell line (IgM, at 0.298 μg per 1 × 106) and the secondary rat anti-mouse IgM-PE/CY7 antibody.

Techniques: Infection

Journal: Vaccines

Article Title: Protective Efficacy Induced by the Common Eimeria Antigen Elongation Factor 2 against Challenge with Three Eimeria Species in Chickens.

doi: 10.3390/vaccines12010018

Figure Lengend Snippet: Figure 4. The proportion of the T cell subpopulation in chickens immunized with PBS, pET-32a tag protein and rEmEF2 was determined by flow cytometry 7 days after the first and second immu- nization. (A) Detection of CD3+CD4+ T lymphocytes in immunized chickens 7 days after the first immunization. (B) Detection of CD3+CD4+ T lymphocytes in immunized chickens 7 days after the second immunization. (C) Detection of CD3+CD8+ T lymphocytes in immunized chickens 7 days after the first immunization. (D) Detection of CD3+CD8+ T lymphocytes in immunized chickens 7 days after the second immunization. 1: PBS control group. 2: pET-32a tag protein control group. 3: rEmEF2 group.

Article Snippet: Whereafter, CD3 mouse anti-chicken FITC antibody (Southern Biotechnology Associates, Birmingham, AL, USA), CD4 mouse anti-chicken PE antibody (Southern Biotechnology Associates, Birmingham, AL, USA) and CD8 mouse anti-chicken PE antibody (Southern Biotechnology Associates, Birmingham, AL, USA) were used to detect the T cell subpopulations with a FACS Calibur flow cytometer (BD Biosciences, Franklin Lakes, NJ, USA).

Techniques: Flow Cytometry, Control