cd25 cell depletion  (Miltenyi Biotec)

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: CD25 + cell depletion was achieved by incubating the PBMCs with 20 μL of FITC-conjugated human anti-chicken CD25 antibody for 20 min at 4°C, followed by an additional 20-min incubation at 4°C with 20 μL of anti-FITC MultiSort MicroBeads antibodies (Miltenyi Biotec, Bergisch Gladbach, North Rhine-Westphalia, Germany).

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: CD25 + cell depletion was achieved by incubating the PBMCs with 20 μL of FITC-conjugated human anti-chicken CD25 antibody for 20 min at 4°C, followed by an additional 20-min incubation at 4°C with 20 μL of anti-FITC MultiSort MicroBeads antibodies (Miltenyi Biotec, Bergisch Gladbach, North Rhine-Westphalia, Germany).

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: CD25 + cell depletion was achieved by incubating the PBMCs with 20 μL of FITC-conjugated human anti-chicken CD25 antibody for 20 min at 4°C, followed by an additional 20-min incubation at 4°C with 20 μL of anti-FITC MultiSort MicroBeads antibodies (Miltenyi Biotec, Bergisch Gladbach, North Rhine-Westphalia, Germany).

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: CD25 + cell depletion was achieved by incubating the PBMCs with 20 μL of FITC-conjugated human anti-chicken CD25 antibody for 20 min at 4°C, followed by an additional 20-min incubation at 4°C with 20 μL of anti-FITC MultiSort MicroBeads antibodies (Miltenyi Biotec, Bergisch Gladbach, North Rhine-Westphalia, Germany).

Techniques: In Vivo, Infection, Injection

Journal: Journal of Clinical Apheresis

Article Title: Suppressive CD8 + T‐Cells Are Key Cellular Mediators of Extracorporeal Photopheresis

doi: 10.1002/jca.70094

Figure Lengend Snippet: Suppressive CD8+ T cells are increased in patients undergoing ECP. For all experiments PBMCs were isolated from whole blood obtained from healthy donors or patients undergoing ECP. (A) Flow cytometry was performed, and CD8+ suppressor cells were identified as indicated. (B–E) CD4+ CD25− cells were isolated from PBMCs, stained with CFSE, and incubated with CD8+ cells at the indicated ratios for 5–7 days. Flow cytometry was performed to identify CFSE dilute CD4+ cells (divided) and the % suppression was calculated by comparing the divided populations to CD4+ cells that were incubated in the absence of CD8+ cells (more details in the materials and methods). (B) CD8‐suppression assays in healthy donors versus ECP patients. (C) CD8‐suppression assays in paired samples obtained prior to the start of ECP (Baseline) or after 1 month of therapy. (D, E) Allo‐suppression assays. CD4+ CD25− cells were isolated from healthy donors or ECP patients. Each donor was paired with an ECP patient to allow for direct comparison between autologous and allogenic suppression results. (D) CD4+ CD25− cells from the healthy donors were incubated with CD8+ cells from the same healthy donor (auto) or a paired ECP patient (allo). (E) CD4+ CD25− cells from the ECP patients were incubated with CD8+ cells from a paired healthy donor (allo) or the same ECP patient (auto). Suppression of cell proliferation was evaluated as described above. Statistics were performed using Student's t ‐test with * indicating p < 0.05, ** indicating p < 0.01, and *** indicating p < 0.001.

Article Snippet: CD4+ CD25− cells were subsequently isolated from the CD8‐depleted cell population by negative selection using the CD4+ T Cell Isolation Kit (Miltenyi Biotec, 130‐096‐533) and CD25 Microbeads (Miltenyi Biotec, 130‐092‐983).

Techniques: Isolation, Flow Cytometry, Staining, Incubation, Comparison