Journal: Scientific Reports

Article Title: Unraveling the role of shrimp hydrolysate as a food supplement in the immune function and fecal microbiota of beagle dogs

doi: 10.1038/s41598-025-09942-8

Figure Lengend Snippet: T lymphocyte proliferation evaluated by flow cytometry. ( A ) Percentage of CD3 + CD4 + cells; ( B ) Percentage of CD3 + CD8 + cells, from dogs fed control diet and experimental diet, that proliferated at least once when non-stimulated (None) or in response to recombinant antigen from Leptospira interrogans (LipL32) and concanavalin A (ConA). Bars correspond to mean plus standard error of the mean. * P < 0.05.

Article Snippet: The PBMC were washed with PBS and stained with anti-dog CD3 FITC-conjugate (Bio-Rad), anti-dog CD4 PE-Cy7-conjugate (eBioscience), and anti-dog CD25 Super Bright 436-conjugate (clone P4A10, 62-0250-42, eBioscience), protected from light, for 25 min at 4 °C.

Techniques: Flow Cytometry, Control, Recombinant

Journal: Scientific Reports

Article Title: Unraveling the role of shrimp hydrolysate as a food supplement in the immune function and fecal microbiota of beagle dogs

doi: 10.1038/s41598-025-09942-8

Figure Lengend Snippet: Intracellular cytokine measurement by flow cytometry. ( A ) Percentage of CD3 + CD4 + cells expressing interferon-gamma (IFN-γ), tumor necrosis factor-alpha (TNF-α) and both cytokines; ( B ) Percentage of CD3 + CD8 + cells expressing IFN-γ, TNF-α and both cytokines; ( C ) Ratio of CD3 + CD4 + and CD3 + CD8 + cells calculated from the percentage of CD4 + and CD8 + T cells; ( D ) Percentage of CD3 + CD4 + CD25 + cells expressing Foxp3 from dogs fed control diet and experimental diet. Bars correspond to mean plus standard error of the mean. ** P < 0.01.

Article Snippet: The PBMC were washed with PBS and stained with anti-dog CD3 FITC-conjugate (Bio-Rad), anti-dog CD4 PE-Cy7-conjugate (eBioscience), and anti-dog CD25 Super Bright 436-conjugate (clone P4A10, 62-0250-42, eBioscience), protected from light, for 25 min at 4 °C.

Techniques: Flow Cytometry, Expressing, Control

Journal: Cell Reports Medicine

Article Title: Cross-organ hierarchy of HLA molecular mismatches in donor-specific antibody development in solid organ transplantations

doi: 10.1016/j.xcrm.2025.102153

Figure Lengend Snippet: Examination of eplet mismatch immunogenicity in mixed lymphocyte reactions The association between the eplet risk score (ERS) and the activation status of responder CD4 + T cells was analyzed. Representative scattergrams are shown in (A), and aggregated scatterplots are shown in (B). Each stimulator-responder pair was analyzed with 3–6 biological replicates. The effect of an anti-55PP antibody (Ab) or an anti-45EV Ab on the percentage of CFSE low CD4 + T cells in the MLR was assessed using a stimulator-responder pair (pair #4, see ). Representative scattergrams are shown in (C), and aggregated data are shown in (D) and (E). Each antibody concentration group was analyzed with 4–12 biological replicates. ∗ p < 0.05. p values were determined using the Wilcoxon rank-sum test, which compared the value at 0 ng/mL Ab with those at 1 × 10 2 and 1 × 10 3 ng/mL Ab. EMn, the number of eplet mismatches; FSC, forward scatter; stim, stimulator. In the scatterplot shown in (B), large dots represent the median value of each stimulator-responder pair, small dots represent values obtained from each individual experiment, vertical lines represent ranges, and the gray area represents 95% confidence interval (CI) of the regression line. In the bar charts shown in (D) and (E), the top of the bar represents the median value, whereas the points represent values from individual experiments.

Article Snippet: Anti-CD4 PE-Cy7 antibody (clone SK3) , Becton, Dickinson and Company , Cat#348789; RRID: AB_400379.

Techniques: Immunopeptidomics, Activation Assay, Concentration Assay

Journal: Frontiers in Veterinary Science

Article Title: Immune cell profile and metabolic preference following intramuscular lipopolysaccharide injection of highly inbred and advanced intercross genetic lines

doi: 10.3389/fvets.2025.1592021

Figure Lengend Snippet: Percentages of (A) Monocyte/Macrophage + , (B) total CD3 + , (C) CD3 + CD1.1 + , (D) CD3 + CD4 + , and (E) CD3 + CD8α + cells isolated from peripheral blood mononuclear cells of genetic line (Ghs, Line-8, Sp-21.1, and AIL-F) ± 1 mg/kg intramuscular LPS injection at baseline, 6 hpi, and 24 hpi. Data represents the mean ± SEM ( n = 10 birds/genetic line at baseline and n = 5/treatment of each genetic line at 6 hpi and 24 hpi). Note that the y-axis is larger in panels (B-E) . Different letter superscripts denote significant differences within a timepoint p ≤ 0.05.

Article Snippet: The flow cytometry panel included the following antibodies to detect innate immune cell and T lymphocyte populations: PE anti-chicken monocyte/macrophage (clone KUL01; mouse IgG1κ), Pacific BlueTM anti-chicken CD3 (clone CT-3; mouse IgG1κ), FITC anti-chicken CD1.1 (clone CB3; mouse IgG1κ), PE/CY7 anti-chicken CD4 (clone CT-4; mouse IgG1κ), Alexa Fluor ® 700 anti-chicken CD8α (clone CT-8; mouse IgG1κ; Southern Biotech, Birmingham, AL).

Techniques: Isolation, Injection