anti chicken cd25 antibody  (Bio-Rad)

Journal: Poultry Science

Article Title: Depletion of CD25 + cells restores Th1, Th2 and Th17 responses and mitigates Eimeria maxima infection in chickens

doi: 10.1016/j.psj.2026.106467

Figure Lengend Snippet: Chicken CD25-specific antiserum preparation, purification, and western blot analysis . (A) The amplification of chicken CD25 gene lane M: DNA Marker DL 2000; lane 1: the amplification products of CD25 gene. (B) Purification of chicken pET-28a-CD25 protein lane M: standard molecular marker for protein; lane 1: purified chicken pET-28a-CD25 protein. (C) Western blot identification of His-tag in chicken pET-28a-CD25 protein lane M: standard molecular marker for protein; lane 1: identification of His-tag in pET-28a-CD25 protein by mouse anti His-tag monoclonal antibody. (D) Western blot identification of chicken CD25 protein from spleen by rat anti-chicken CD25 polyclonal antibody lane M: standard molecular marker for protein; line 1: identification of CD25 protein by rat anti-chicken CD25 polyclonal antibody (∼23.5 kDa); line 2: identification of CD25 protein by negative rat serum.

Article Snippet: FITC-conjugated human anti-chicken CD25 antibody was sourced from BIO-RAD (Hercules, CA, USA), while PE-conjugated mouse anti-chicken CD4 antibody was obtained from Southern Biotechnology Associates (Birmingham, AL, USA).

Techniques: Purification, Western Blot, Amplification, Marker

Journal: Poultry Science

Article Title: Depletion of CD25 + cells restores Th1, Th2 and Th17 responses and mitigates Eimeria maxima infection in chickens

doi: 10.1016/j.psj.2026.106467

Figure Lengend Snippet: Dynamic changes in chicken CD4 + CD25 + T cells in PBMCs and splenocytes of E. maxima -infected chickens. The flow cytometry serial gating strategy for detection of the percentage of CD4 + CD25 + T cells/CD4 + T cells in PBMCs (A) and splenocytes (B). (C) The percentage of CD4 + CD25 + T cells/CD4 + T cells in PBMCs at 0, 3, 7, 11, 14 and 21 days post-infection. (D) The percentage of CD4 + CD25 + T cells/CD4 + T cells in splenocytes at 0, 3, 7, 11, 14 and 21 days post-infection. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001.

Article Snippet: FITC-conjugated human anti-chicken CD25 antibody was sourced from BIO-RAD (Hercules, CA, USA), while PE-conjugated mouse anti-chicken CD4 antibody was obtained from Southern Biotechnology Associates (Birmingham, AL, USA).

Techniques: Infection, Flow Cytometry

Journal: Poultry Science

Article Title: Depletion of CD25 + cells restores Th1, Th2 and Th17 responses and mitigates Eimeria maxima infection in chickens

doi: 10.1016/j.psj.2026.106467

Figure Lengend Snippet: Effect of CD25 + cells depletion in vitro on the immunomodulatory function of PBMCs infected with E. maxima . (A) The percentage of CD25 + cells in chicken PBMCs before depletion in vitro . (B) The percentage of CD25 + cells in chicken PBMCs after depletion in vitro . (C) The effect of E. maxima sporozoite protein stimulation on the cell proliferation in PBMCs depleted with CD25 + cells in vitro . (D-J) The effects of E. maxima sporozoite protein stimulation on the mRNA levels of cytokines and CTLA-4 in PBMCs depleted with CD25 + cells in vitro . * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001.

Article Snippet: FITC-conjugated human anti-chicken CD25 antibody was sourced from BIO-RAD (Hercules, CA, USA), while PE-conjugated mouse anti-chicken CD4 antibody was obtained from Southern Biotechnology Associates (Birmingham, AL, USA).

Techniques: In Vitro, Infection

Journal: Poultry Science

Article Title: Depletion of CD25 + cells restores Th1, Th2 and Th17 responses and mitigates Eimeria maxima infection in chickens

doi: 10.1016/j.psj.2026.106467

Figure Lengend Snippet: Interference impact of CD25 + cells depletion in vivo on the immunomodulatory function of PBMCs infected with E. maxima . (A) Dynamic changes of the percentage of CD4 + CD25 + T cells/CD4 + T cells in PBMCs injected intravenously with rat anti-chicken CD25 polyclonal antibody. (B) The effects of E. maxima sporozoite protein stimulation on the cell proliferation in PBMCs blocked with CD25 molecule in vivo . (C-I) The effects of E. maxima sporozoite protein stimulation on the mRNA levels of cytokines and CTLA-4 in PBMCs blocked with CD25 molecule in vivo . * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001.

Article Snippet: FITC-conjugated human anti-chicken CD25 antibody was sourced from BIO-RAD (Hercules, CA, USA), while PE-conjugated mouse anti-chicken CD4 antibody was obtained from Southern Biotechnology Associates (Birmingham, AL, USA).

Techniques: In Vivo, Infection, Injection

Journal: BMC Veterinary Research

Article Title: Immune regulation and transcriptomic profiling of chicken CD8⁺ T cells in response to infection with the IBV strain K047-12 during in vitro culture

doi: 10.1186/s12917-026-05284-3

Figure Lengend Snippet: Immune cell composition in the chicken splenocytes during ConA + IL-2 stimulation and IBV infection. Chicken splenocytes were stimulated with Concanavalin A (ConA) and chicken IL-2 for 48 h, followed by IBV infection for an additional 48 h. A Flow cytometric analysis of immune cell phenotypes in splenocytes before and after ConA + IL-2 stimulation. Populations of CD3⁺ T cells, CD8⁺ T cells, CD4⁺ T cells, B cells (Bu-1⁺), and macrophages (Mac/Mono⁺ and MHC II⁺) are shown (n = 5). Statistical significance was determined with a paired t-test (*p < 0.05). B Cell morphology was monitored from 48 h (0 h post-IBV infection) to 96 h (48 h post- infection), and representative graphs show changes in cell confluency recorded every 12 h after IBV infection. The experiments are performed in duplicate. C Morphological changes in splenocytes were assessed at three time points—0 h (unstimulated), 48 h (stimulated with ConA + IL-2), and 96 h (stimulated with ConA + IL-2 for 48 h followed by IBV infection for 48 h) using Diff-Quik staining. Representative images are shown (n = 5) ( D ) Cell viability following IBV infection for 48 h, evaluated by flow cytometry using LIVE/DEAD™ fixable dye. ( E , F ) The cultured cells in the absence (Control) or presence of infectious bronchitis virus (IBV) were harvested and analyzed by flow cytometry for immune cell phenotypic changes. The frequencies of CD4⁺, CD8⁺, CD44⁺ CD4⁺, and CD44⁺ CD8⁺ cells within live cells were assessed in IBV-infected and uninfected control groups. (G) The frequencies of CD25high populations were assessed within CD4⁺ and CD44⁺ CD4⁺ T cells, as well as within CD8⁺ and CD44⁺ CD8⁺ T cells. The mean fluorescence intensity (MFI) of CD25 within CD44⁺ CD4⁺ and CD44⁺ CD8⁺ T cells was also evaluated. Representative flow cytometry plots are shown. Data are pooled from 5–6 independent experiments. Statistical significance was determined with a paired t-test (* p < 0.05, **p < 0.001)

Article Snippet: Chicken CD25 , FITC , AbD13504 , HCA173F , Bio-Rad (USA).

Techniques: Infection, Diff-Quik, Staining, Flow Cytometry, Cell Culture, Control, Virus, Fluorescence

Journal: Veterinary research

Article Title: Immunomodulatory effects of Eimeria maxima surface antigen (EmSAG) as an IFN-γ inhibitory molecule on peripheral blood mononuclear cells (PBMCs) and T cell subsets in chickens.

doi: 10.1186/s13567-025-01535-7

Figure Lengend Snippet: Figure 6 Detection of cell sorting purity of T cell subsets in chicken by flow cytometry. A: Magnetic bead sorting of chicken CD8+ T cells. B: Magnetic bead sorting of chicken CD4+ T cells. C: Magnetic bead sorting of chicken CD4+CD25− T cells. D: Magnetic bead sorting of chicken CD4+CD25+ T cells. 1: Lymphocytes in chicken peripheral blood. 2: Singlet cells. 3: Blank cells incubated without fluorescent antibody. 4: CD8+, CD4+, CD4+CD25−, and CD25+ cells before cell sorting. 5: CD8+, CD4+, CD4+CD25−, and CD25+ cells after cell sorting. 6: CD4+CD25+ T cells before cell sorting. 7: CD4+CD25+ T cells after cell sorting.

Article Snippet: In a similar manner, chicken PBMCs were incubated with FITC-conjugated human anti-chicken CD25 antibodies (Bio-Rad, Hercules, CA, USA) and anti-FITC MultiSort MicroBeads antibodies (Miltenyi Biotec, Bergisch Gladbach, North Rhine-Westphalia, Germany) for 30 min at 4 °C.

Techniques: FACS, Flow Cytometry, Incubation

Journal: Veterinary research

Article Title: Immunomodulatory effects of Eimeria maxima surface antigen (EmSAG) as an IFN-γ inhibitory molecule on peripheral blood mononuclear cells (PBMCs) and T cell subsets in chickens.

doi: 10.1186/s13567-025-01535-7

Figure Lengend Snippet: Figure 7 Effects of rEmSAG on cytokines transcription of T cell subsets in chicken. A: Relative fold changes in IFN-γ, IL-2, IL-4, IL-10, and TGF-β1 transcription were defined in chicken CD4+ T cells following stimulation with 20 μg/mL rEmSAG. B: Relative fold changes in IFN-γ, IL-2, and TNF-α transcription were defined in chicken CD8+ T cells following stimulation with 20 μg/mL rEmSAG. C: Relative fold changes in IL-10, TGF-β1, and CTLA-4 transcription were defined in chicken CD4+CD25+ T cells following stimulation with 20 μg/mL rEmSAG.

Article Snippet: In a similar manner, chicken PBMCs were incubated with FITC-conjugated human anti-chicken CD25 antibodies (Bio-Rad, Hercules, CA, USA) and anti-FITC MultiSort MicroBeads antibodies (Miltenyi Biotec, Bergisch Gladbach, North Rhine-Westphalia, Germany) for 30 min at 4 °C.

Techniques:

Journal: Veterinary research

Article Title: Immunomodulatory effects of Eimeria maxima surface antigen (EmSAG) as an IFN-γ inhibitory molecule on peripheral blood mononuclear cells (PBMCs) and T cell subsets in chickens.

doi: 10.1186/s13567-025-01535-7

Figure Lengend Snippet: Figure 8 Effects of rEmSAG on cytokines transcription of chicken CD4+CD25− T cells following CD25+ cell depletion. qPCR was used to detect the transcription of cytokines including IFN-γ, IL-2, IL-4, IL-10, and TGF-β1 in chicken CD4+CD25− T cells stimulated with 20 μg/mL rEmSAG following the depletion of CD25+ cells from CD4+ T cells.

Article Snippet: In a similar manner, chicken PBMCs were incubated with FITC-conjugated human anti-chicken CD25 antibodies (Bio-Rad, Hercules, CA, USA) and anti-FITC MultiSort MicroBeads antibodies (Miltenyi Biotec, Bergisch Gladbach, North Rhine-Westphalia, Germany) for 30 min at 4 °C.

Techniques: