Journal: iScience
Article Title: Characterization of human CD34 + HSPC-derived neutrophils with limited myeloid-derived immunosuppressive cell activity
doi: 10.1016/j.isci.2025.113404
Figure Lengend Snippet: CD16 + cultured neutrophils were similar to PMNs in mobility, ROS production, phagocytosis, and microbial killing (A) Boxplots and violin plots showing the relative expression of genes and proteins involved in neutrophil effector functions . (B) CD11b/CD18-mediated adhesion assay of PMNs (blue) and SL-II CD16 + neutrophils (pink) to uncoated plastic plates ( n = 4). (C) Chemotactic potential of fluorescently labeled PMNs (blue) and SL-II CD16 + neutrophils (pink) based on movement through filters with a pore size of 3 micron ( n = 5). (D) NADPH oxidase assay to determine the production of extracellular peroxide after the addition of opsonized E. coli , zymosan, STZ, PMA, and PAF/fMLP of PMNs (blue) and SL-II CD16 + neutrophils (pink) ( n = 4 for SL-II CD16 + neutrophils and n = 8 for PMNs). (E) Phagocytosis of either unopsonized or opsonized zymosan by PMNs (blue) versus SL-II CD16 + neutrophils (pink) measured by flow cytometry ( n = 3 for SL-II CD16 + neutrophils and n = 4 for PMNs). (F) Representative images of phagocytosis of either unopsonized or opsonized zymosan labeled with FITC solution (green) by PMNs labeled with calcein red-orange (orange) versus SL-II CD16 + neutrophils labeled with calcein red-orange (orange) at the latest timepoint as visualized by imaging flow cytometric analysis. Scale bar was set at 10 μm ( n = 3). (G and H) Killing of opsonized E. coli and S. aureus, and unopsonized and opsonized C. albicans, respectively, shown for PMNs (blue) ( n = 3) versus SL-II CD16 + neutrophils (pink) ( n = 3). Killing was quantified as the inverse of colony-forming units (CFU) as a percentage relative to the CFU at the start of the assay. Negative values, signifying an increase in the number of colonies were considered to be 0. Data shown in (A–D, H) is represented as median and interquartile range, and (E) and (G) is represented as mean ± SD. p values were calculated using Mann-Whitney U tests and labeled as ∗ p < 0.05 and ∗∗ p < 0.01. n values represent the number of individual donor samples.
Article Snippet: Mouse Anti-Human CD66b Monoclonal antibody, FITC Conjugated (Clone 80H3) , Bio-Rad , Cat #MCA216F; RRID: AB_2077860.
Techniques: Cell Culture, Expressing, Cell Adhesion Assay, Labeling, Pore Size, Flow Cytometry, Imaging, MANN-WHITNEY
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