Journal: Technology and Health Care

Article Title: Prediction and identification of epitopes in the Echinococcus multilocularis thrombospondin 3 antigen

doi: 10.3233/THC-212983

Figure Lengend Snippet: The enzyme-linked immunosorbent assay test on the cytokines of mouse spleen lymphocytes immunized with thrombospondin 3. A: The interleukin-2 test; B: the interferon-y test; C: the tumor necrosis factor- β test; D: the interleukin-4 test; E: the interleukin-17 test. P * ⁣ * * < 0.001.

Article Snippet: PE-labeled anti-mouse IL-4 flow antibody , BD, USA.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Technology and Health Care

Article Title: Prediction and identification of epitopes in the Echinococcus multilocularis thrombospondin 3 antigen

doi: 10.3233/THC-212983

Figure Lengend Snippet: The enzyme-linked immune absorbent spot test of the cytokines of mouse spleen lymphocytes immunized with thrombospondin 3. A: The interleukin-2 test; B: the interferon- γ test; C: the tumor necrosis factor- β test; D: the interleukin-4 test. P * ⁣ * * < 0.001.

Article Snippet: PE-labeled anti-mouse IL-4 flow antibody , BD, USA.

Techniques: Spot Test

Journal: Technology and Health Care

Article Title: Prediction and identification of epitopes in the Echinococcus multilocularis thrombospondin 3 antigen

doi: 10.3233/THC-212983

Figure Lengend Snippet: Experimental reagents

Article Snippet: PE-labeled anti-mouse IL-4 flow antibody , BD, USA.

Techniques: Cell Culture, Labeling, Enzyme-linked Immunosorbent Assay, Enzyme-linked Immunospot

Journal:

Article Title: Role of Natural Killer Cells in Resistance against Friend Retrovirus-Induced Leukemia

doi: 10.1128/JVI.75.7.3152-3163.2001

Figure Lengend Snippet: Changes in the numbers of TER-119+ erythroid cells (a), F-MuLV gp70-expressing cells detected with MAb 720 (b), and CD4+ (c) and CD8+ (d) cells in the spleens of mice inoculated with FV. CB6F1 mice were either immunized with peptide i (○) or given CFA without a peptide (●). Four weeks later, they were inoculated with 150 SFFU of FV. A group of three or four animals were killed at each indicated point, and their spleen cells were subjected to flow-cytometric analyses. Data presented here are means ± SEM. The dashed line in panel b indicates the limit of detection by the flow-cytometric analysis.

Article Snippet: Antibodies and their final concentrations used in the present study were as follows: fluorescein isothiocyanate (FITC)-conjugated anti-mouse CD4 (rat immunoglobulin G2b [IgG2b]; Seikagaku Corporation, Tokyo, Japan) at 0.5 μg/10 6 cells, phycoerythrin (R-PE)-conjugated anti-mouse CD8 (rat IgG2a; Caltag Laboratories, Burlingame, Calif.) at 1 μg/10 6 cells, FITC-conjugated anti-mouse CD69 (hamster IgG; PharMingen, San Diego, Calif.) at 1 μg/10 6 cells, R-PE-conjugated anti-mouse B220 (rat IgG2a; Coulter Immunology, Hialeah, Fla.) at 0.5 μg/10 6 cells, FITC-conjugated anti-NK1.1 (mouse IgG2a; PharMingen) at 2 μg/10 6 cells, biotin-conjugated anti-mouse Pan-NK (DX5, rat IgM; PharMingen) at 1 μg/10 6 cells, and allophycocyanin-conjugated anti-mouse TER-119 (PharMingen) at 0.2 μg/10 6 cells.

Techniques: Expressing

Journal:

Article Title: Role of Natural Killer Cells in Resistance against Friend Retrovirus-Induced Leukemia

doi: 10.1128/JVI.75.7.3152-3163.2001

Figure Lengend Snippet: Detection of cytotoxic effector cells in FV-infected CB6F1 mice. Mice were either immunized with 10 μg of peptide i/mouse or given CFA emulsion without a peptide. B220− spleen cells were separated into CD8+, CD4+, and CD4− CD8− populations, and their cytotoxic activities against FBL-3 (○), Y57-2C (□), and EL-4 (●) cells were tested by incubating the effector and labeled target cells for 12 h. Representative data obtained from a set of experiments performed at PID 9 are shown here, and the results obtained from the six repeated experiments were consistent with these charts.

Article Snippet: Antibodies and their final concentrations used in the present study were as follows: fluorescein isothiocyanate (FITC)-conjugated anti-mouse CD4 (rat immunoglobulin G2b [IgG2b]; Seikagaku Corporation, Tokyo, Japan) at 0.5 μg/10 6 cells, phycoerythrin (R-PE)-conjugated anti-mouse CD8 (rat IgG2a; Caltag Laboratories, Burlingame, Calif.) at 1 μg/10 6 cells, FITC-conjugated anti-mouse CD69 (hamster IgG; PharMingen, San Diego, Calif.) at 1 μg/10 6 cells, R-PE-conjugated anti-mouse B220 (rat IgG2a; Coulter Immunology, Hialeah, Fla.) at 0.5 μg/10 6 cells, FITC-conjugated anti-NK1.1 (mouse IgG2a; PharMingen) at 2 μg/10 6 cells, biotin-conjugated anti-mouse Pan-NK (DX5, rat IgM; PharMingen) at 1 μg/10 6 cells, and allophycocyanin-conjugated anti-mouse TER-119 (PharMingen) at 0.2 μg/10 6 cells.

Techniques: Infection, Labeling

Journal:

Article Title: Role of Natural Killer Cells in Resistance against Friend Retrovirus-Induced Leukemia

doi: 10.1128/JVI.75.7.3152-3163.2001

Figure Lengend Snippet: In vivo depletion of NK cell activity by injection of anti-asialo-GM1 Ab. (a and b) CB6F1 mice immunized with peptide i were injected either with 60 μg of anti-asialo-GM1 Ab each (b) or with normal rabbit serum (a) and were infected with FV. Spleen cells were obtained at PID 9, and the NK cell activity of the B220− population was tested by using YAC-1 (▵) and EL-4 (●) target cells. Data from two separate experiments are shown together here. Injection of higher doses of anti-asialo-GM1 Ab gave the same results when B200− cells were similarly tested for their YAC-1-killing activities. (c and d) Flow cytometric analyses for the expression of the NK cell markers on spleen cells obtained from mice injected with normal rabbit serum (c) or anti-asialo-GM1 Ab (d). Experiments were performed twice and gave essentially the same results as those shown here. (e through j) Cytotoxicity assays using different cell populations isolated from spleen B220− cells of peptide-immunized, FV-infected mice. CD8+, CD4+, and CD4− CD8− populations were purified as described for the experiments shown in Fig. ​Fig.33 from CB6F1 mice injected with anti-asialo-GM1 Ab (f, h, and j) or from those injected with control rabbit serum (e, g, and i). The experiments were performed twice at PID 7 and 9, and the results from the repeated experiments were consistent with the representative data shown here. Target cells used were YAC-1 (Δ), FBL-3 (○), and EL-4 (●).

Article Snippet: Antibodies and their final concentrations used in the present study were as follows: fluorescein isothiocyanate (FITC)-conjugated anti-mouse CD4 (rat immunoglobulin G2b [IgG2b]; Seikagaku Corporation, Tokyo, Japan) at 0.5 μg/10 6 cells, phycoerythrin (R-PE)-conjugated anti-mouse CD8 (rat IgG2a; Caltag Laboratories, Burlingame, Calif.) at 1 μg/10 6 cells, FITC-conjugated anti-mouse CD69 (hamster IgG; PharMingen, San Diego, Calif.) at 1 μg/10 6 cells, R-PE-conjugated anti-mouse B220 (rat IgG2a; Coulter Immunology, Hialeah, Fla.) at 0.5 μg/10 6 cells, FITC-conjugated anti-NK1.1 (mouse IgG2a; PharMingen) at 2 μg/10 6 cells, biotin-conjugated anti-mouse Pan-NK (DX5, rat IgM; PharMingen) at 1 μg/10 6 cells, and allophycocyanin-conjugated anti-mouse TER-119 (PharMingen) at 0.2 μg/10 6 cells.

Techniques: In Vivo, Activity Assay, Injection, Infection, Expressing, Isolation, Purification

Journal:

Article Title: Role of Natural Killer Cells in Resistance against Friend Retrovirus-Induced Leukemia

doi: 10.1128/JVI.75.7.3152-3163.2001

Figure Lengend Snippet: Cytotoxic activity of a CD4+ T-cell clone, SB14-31, specific for an F-MuLV env-encoded epitope. (a) SB14-31 cells were incubated with various target cells with or without preincubation with peptide fn. 51Cr release during 4 h of incubation at an E:T ratio of 20 was measured. (b) LB 27.4 target cells were either incubated with the indicated Ab-binding peptides after 51Cr labeling or infected with the indicated recombinant vaccinia virus for 16 h at a multiplicity of infection of 10 and then labeled. Pretreated LB 27.4 cells were then incubated with SB14-31 cells for 4 h at an E:T ratio of 20:1. Experiments were performed at least twice at various E:T ratios, and the results were consistent with the representative data shown here.

Article Snippet: Antibodies and their final concentrations used in the present study were as follows: fluorescein isothiocyanate (FITC)-conjugated anti-mouse CD4 (rat immunoglobulin G2b [IgG2b]; Seikagaku Corporation, Tokyo, Japan) at 0.5 μg/10 6 cells, phycoerythrin (R-PE)-conjugated anti-mouse CD8 (rat IgG2a; Caltag Laboratories, Burlingame, Calif.) at 1 μg/10 6 cells, FITC-conjugated anti-mouse CD69 (hamster IgG; PharMingen, San Diego, Calif.) at 1 μg/10 6 cells, R-PE-conjugated anti-mouse B220 (rat IgG2a; Coulter Immunology, Hialeah, Fla.) at 0.5 μg/10 6 cells, FITC-conjugated anti-NK1.1 (mouse IgG2a; PharMingen) at 2 μg/10 6 cells, biotin-conjugated anti-mouse Pan-NK (DX5, rat IgM; PharMingen) at 1 μg/10 6 cells, and allophycocyanin-conjugated anti-mouse TER-119 (PharMingen) at 0.2 μg/10 6 cells.

Techniques: Activity Assay, Incubation, Binding Assay, Labeling, Infection, Recombinant

Journal:

Article Title: Role of Natural Killer Cells in Resistance against Friend Retrovirus-Induced Leukemia

doi: 10.1128/JVI.75.7.3152-3163.2001

Figure Lengend Snippet: Cytotoxic activities of four different CD4+ T-cell clones specific for peptide i. T-cell clones F5-5 (a), FP3-10 (b), FP8-7 (c), and FP10-16 (d) were tested for their ability to lyse LB 27.4 target cells by incubation at the indicated E:T ratios for 3 h. LB 27.4 cells were incubated either with peptide i (□, ○, ▵) or with the control peptide of the same length, ie (●). Killing assays were performed in the absence (○, ●) or presence of anti-CD4 (□) or anti-CD8 (▵) MAb. Assays were performed at least twice, and the results were consistent with the representative data shown here.

Article Snippet: Antibodies and their final concentrations used in the present study were as follows: fluorescein isothiocyanate (FITC)-conjugated anti-mouse CD4 (rat immunoglobulin G2b [IgG2b]; Seikagaku Corporation, Tokyo, Japan) at 0.5 μg/10 6 cells, phycoerythrin (R-PE)-conjugated anti-mouse CD8 (rat IgG2a; Caltag Laboratories, Burlingame, Calif.) at 1 μg/10 6 cells, FITC-conjugated anti-mouse CD69 (hamster IgG; PharMingen, San Diego, Calif.) at 1 μg/10 6 cells, R-PE-conjugated anti-mouse B220 (rat IgG2a; Coulter Immunology, Hialeah, Fla.) at 0.5 μg/10 6 cells, FITC-conjugated anti-NK1.1 (mouse IgG2a; PharMingen) at 2 μg/10 6 cells, biotin-conjugated anti-mouse Pan-NK (DX5, rat IgM; PharMingen) at 1 μg/10 6 cells, and allophycocyanin-conjugated anti-mouse TER-119 (PharMingen) at 0.2 μg/10 6 cells.

Techniques: Clone Assay, Incubation

Journal:

Article Title: Role of Natural Killer Cells in Resistance against Friend Retrovirus-Induced Leukemia

doi: 10.1128/JVI.75.7.3152-3163.2001

Figure Lengend Snippet: In vivo depletion of asialo-GM1+ cells and its effect on T cells and protective immunity against FV infection induced by peptide immunization. (a through d) Mice used for the experiments whose results are shown in Fig. ​Fig.77 were also analyzed for the presence of CD4+ and CD8+ T cells in the spleen and their ability to mount viral-antigen-specific CD4+ T-cell responses. Flow-cytometric analyses for the expression of CD4 and CD8 were performed by using pooled whole spleen cells obtained from the mice injected with anti-asialo-GM1 Ab (b) or normal rabbit serum (a). Experiments were performed twice at PID 7 and 9, and results obtained from the repeated experiments were consistent with the representative data shown here. Numbers indicate percentages of CD4+ and CD8+ cells among live nucleated spleen cells. B220− CD8− CD4+ T cells purified for the experiments whose results are shown in Fig. ​Fig.7g7g and h were also tested for their proliferative activities in response to stimulation with peptide i. CD4+ T cells purified from the mice injected with anti-asialo-GM1 Ab (d) and those purified from control mice given normal rabbit serum (c) were incubated with X-irradiated syngeneic spleen cells and the indicated amount of peptide i (○). As controls, the CD4+ T cells purified from the anti-asialo-GM1 Ab-injected mice were also stimulated with an endogenous retroviral env-derived peptide ie (●) and the influenza virus nucleoprotein-derived peptide NP366–374 (▵). Experiments were performed twice, and results obtained from the repeated experiments were consistent with the representative data shown here. (e) Development of FV-induced leukemia in CB6F1 mice immunized with peptide i. Mice were either immunized with 10 μg of peptide i each (○, ▵, □) or given CFA alone (●). Two groups of the immunized mice were then injected with anti-asialo-GM1 Ab (▵) or control rabbit serum (□), while the remaining group (○) was not injected with any Ab. All mice were inoculated with 150 SFFU of FV.

Article Snippet: Antibodies and their final concentrations used in the present study were as follows: fluorescein isothiocyanate (FITC)-conjugated anti-mouse CD4 (rat immunoglobulin G2b [IgG2b]; Seikagaku Corporation, Tokyo, Japan) at 0.5 μg/10 6 cells, phycoerythrin (R-PE)-conjugated anti-mouse CD8 (rat IgG2a; Caltag Laboratories, Burlingame, Calif.) at 1 μg/10 6 cells, FITC-conjugated anti-mouse CD69 (hamster IgG; PharMingen, San Diego, Calif.) at 1 μg/10 6 cells, R-PE-conjugated anti-mouse B220 (rat IgG2a; Coulter Immunology, Hialeah, Fla.) at 0.5 μg/10 6 cells, FITC-conjugated anti-NK1.1 (mouse IgG2a; PharMingen) at 2 μg/10 6 cells, biotin-conjugated anti-mouse Pan-NK (DX5, rat IgM; PharMingen) at 1 μg/10 6 cells, and allophycocyanin-conjugated anti-mouse TER-119 (PharMingen) at 0.2 μg/10 6 cells.

Techniques: In Vivo, Infection, Expressing, Injection, Purification, Incubation, Irradiation, Derivative Assay

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 m onuclear cell binding spe ficity. (A): Histopaque 1077-treated PBMC (1 × 106 cells) with no added antibodies (negative control); (B): PBMC stained with mouse antichicken 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.

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 m onuclear cell binding spe ficity. (A): Histopaque 1077-treated PBMC (1 × 106 cells) with no added antibodies (negative control); (B): PBMC stained with mouse antichicken 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.

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 m onuclear cell binding spe ficity. (A): Histopaque 1077-treated PBMC (1 × 106 cells) with no added antibodies (negative control); (B): PBMC stained with mouse antichicken 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.

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 m onuclear cell binding spe ficity. (A): Histopaque 1077-treated PBMC (1 × 106 cells) with no added antibodies (negative control); (B): PBMC stained with mouse antichicken 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.

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 m onuclear cell binding spe ficity. (A): Histopaque 1077-treated PBMC (1 × 106 cells) with no added antibodies (negative control); (B): PBMC stained with mouse antichicken 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.

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 m onuclear cell binding spe ficity. (A): Histopaque 1077-treated PBMC (1 × 106 cells) with no added antibodies (negative control); (B): PBMC stained with mouse antichicken 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.

Techniques: Infection

Journal: ImmunoHorizons

Article Title: Characterization of Canine Peyer’s Patches by Multidimensional Analysis: Insights from Immunofluorescence, Flow Cytometry, and Single-Cell RNA Sequencing

doi: 10.4049/immunohorizons.2300091

Figure Lengend Snippet: Structural organization of canine Peyer’s patches, according to immunofluorescence data. ( A ) Sample collection and preparation. Intestinal tissues from two dogs were sampled, and regions of ∼2 cm in length were identified as Peyer’s patches (PPs). The PPs were isolated, sectioned on a cryostat, and prepared for microscopy, with the various structures within the intestine identified before DAPI staining. ( B ) Visualization of PPs with H&E show different staining intensities within the follicles. The staining with Ki67, for proliferating cells, and CD21, for B cells, allows the visualization of proliferating B cells within the GC. ( C ) Immunofluorescence staining of PPs. PPs were stained with DAPI in combination with Abs against CD8 and one of the following markers: CD21, CD4, or FOXP3. Epifluorescence microscopy is shown at original magnification ×10, visualizing specific structures, including the subepithelial dome (SED), T cell areas, and B cell follicles. ( D ) Myeloid cell staining. Immunostaining of the CD14 and CD11c markers, along with CD21, was used to identify dendritic cells within the SED. Different images from the lower and upper parts of the sample were captured to adequately show the CD11c staining in proximity to both the epithelium and the follicle.

Article Snippet: We used the following Abs: mouse anti-dog CD3 Alexa Fluor 700 (Bio-Rad, MCA1774A700), rat anti-dog CD4 PE-Cy7 (Bio-Rad, MCA1038PeCy7), rat anti-dog CD8 Pacific Blue (Bio-Rad, MCA1039PB), and mouse anti-dog CD21 Alexa Fluor 647 (Bio-Rad, MCA1781A647).

Techniques: Immunofluorescence, Isolation, Microscopy, Staining, Epifluorescence Microscopy, Immunostaining

Journal: ImmunoHorizons

Article Title: Characterization of Canine Peyer’s Patches by Multidimensional Analysis: Insights from Immunofluorescence, Flow Cytometry, and Single-Cell RNA Sequencing

doi: 10.4049/immunohorizons.2300091

Figure Lengend Snippet: Characterization of T and B cell populations in canine Peyer’s patches by flow cytometry. ( A ) Peyer’s patches (PPs) from two dogs were dissociated, and single-cell suspensions were stained for flow cytometry in a Cytek Northern Lights flow cytometer. ( B ) Proportions of T and B cells. T cells accounted for 41.9% and B cells for 22.2% of the cell population analyzed. Within the B cell population, 11.6% of the cells were positive for Bcl-6. T cells were further separated into CD4 + , CD8 + , and CD4 + CD8 + subsets, revealing high levels of heterogeneity in the expression of the regulatory marker FOXP3. ( C ) FOXP3 expression in T cell subsets. The percentage of FOXP3 + cells was significantly higher among CD3 + CD4 + CD8 + cells than among CD3 + CD4 + or CD3 + CD8 + cells. Statistical analysis was performed by two-way ANOVA. * p ≤ 0.05, **** p ≤ 0.001. dp, double-positive.

Article Snippet: We used the following Abs: mouse anti-dog CD3 Alexa Fluor 700 (Bio-Rad, MCA1774A700), rat anti-dog CD4 PE-Cy7 (Bio-Rad, MCA1038PeCy7), rat anti-dog CD8 Pacific Blue (Bio-Rad, MCA1039PB), and mouse anti-dog CD21 Alexa Fluor 647 (Bio-Rad, MCA1781A647).

Techniques: Flow Cytometry, Staining, Northern Blot, Expressing, Marker

Journal: ImmunoHorizons

Article Title: Characterization of Canine Peyer’s Patches by Multidimensional Analysis: Insights from Immunofluorescence, Flow Cytometry, and Single-Cell RNA Sequencing

doi: 10.4049/immunohorizons.2300091

Figure Lengend Snippet: Gene expression distribution, B and T cell subpopulations, and the Peyer’s patch atlas. ( A ) Marker expression. The expression of markers such as ICOS, TOP2A, JCHAIN, CCR10, and RORC was visualized across the clusters, facilitating cell subtype identification. ( B ) T cell subpopulations. The distribution of different T cell subpopulations is presented as a pie chart, with the corresponding percentages. ( C ) B cell subpopulations. The distribution of different B cell subpopulations is shown as a pie chart, with the corresponding percentages. ( D ) We propose an annotation of the immune cells identified within clusters, based on the specific expression patterns of marker genes. Bcl-6, B cell lymphoma 6; CD62L, CD62 L-selectin; DZ, dark zone; GC, germinal center; ILC3, innate lymphoid cells group 3; LZ, light zone; PD-1, programmed cell death protein 1; Tfh, T follicular helper cell.

Article Snippet: We used the following Abs: mouse anti-dog CD3 Alexa Fluor 700 (Bio-Rad, MCA1774A700), rat anti-dog CD4 PE-Cy7 (Bio-Rad, MCA1038PeCy7), rat anti-dog CD8 Pacific Blue (Bio-Rad, MCA1039PB), and mouse anti-dog CD21 Alexa Fluor 647 (Bio-Rad, MCA1781A647).

Techniques: Gene Expression, Marker, Expressing

Journal: Animals : an Open Access Journal from MDPI

Article Title: Evaluation of a Novel Precision Biotic on Enterohepatic Health Markers and Growth Performance of Broiler Chickens under Enteric Challenge

doi: 10.3390/ani12192502

Figure Lengend Snippet: Target phenotypes used in analysis of intestinal T cells.

Article Snippet: CD4 , T-helper cells , MCA2164F Biorad.

Techniques: Marker