mouse anti human cd66b fitc  (SouthernBiotech)

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

Journal: Cancer Immunology, Immunotherapy

Article Title: HPV-associated head and neck cancer is characterized by distinct profiles of CD8 + T cells and myeloid-derived suppressor cells

doi: 10.1007/s00262-023-03571-8

Figure Lengend Snippet: PMN MDSC fractions and CD8 + T cell ratios differ in HPV + and HPV − patients and are associated with survival in the HPV + cohort. Analysis of 61 tumor tissue samples from patients with p16 + or p16 − oropharyngeal cancer (second cohort, see Table ) by multi-color immunofluorescence. The frequency and density of CD66b +/ LOX1 + PMN-MDSC ( A ) and frequency of Granzyme B + cytotoxic or Ki67 + proliferating ( B ) CD8 + T cells was determined by digital pathology using tissue studio software modules. ( C ) shows PMN-MDSC to effector T cell ratios. The prognostic value of the frequency of PMN-MDSC was analyzed in HPV + and HPV − patients using Kaplan–Meier curves and overall survival ( D ). Data are presented as median dot plots. The Mann–Whitney rank sum test and the Kruskal–Wallis ANOVA on ranks [post hoc: Dunn‘s method] were used for statistical analysis. Survival analysis is depicted in Kaplan–Meier curves. * indicates a p value < 0.05

Article Snippet: Subsequently, samples were incubated with goat anti-mouse IgG (H + L) Alexa Flour 405 and donkey anti-rabbit IgG (H + L) Alexa 546 (both, ThermoFisher scientific) for 45 min followed by 60-min mouse anti-human CD66b FITC (clone 80H3, Beckmann coulter, Krefeld, Germany) and CD8 Alexa Flour 647 (clone LT8, Bio-Rad, München, Germany).

Techniques: Immunofluorescence, Software, MANN-WHITNEY

Journal: Bioactive Materials

Article Title: A culture model to analyze the acute biomaterial-dependent reaction of human primary neutrophils in vitro ☆

doi: 10.1016/j.bioactmat.2022.05.036

Figure Lengend Snippet: Neutrophil purity assessment, activity and cell survival on different material. A) Flow cytometry histograms showing CD66b and CD16, and CD14 expression on neutrophils. B) Representative images of live/dead staining after 5 and 24 h showing living cells in green with dead cells in red. C) Metabolic activity of neutrophils measured via cell titer blue assay and expressed as fluorescence units over time for each material. D) LDH release by neutrophils cultured on different materials expressed as a percentage of the maximum amount of LDH release after cell lysis. E) The percentage of living cells after 5 h. F) The percentage of alive cells after 24 h. Each bar represents the mean of 5 donors + SD. Abbreviations: LDH, lactate dehydrogenase. a = statistically significant to THA, b = statistically significant to THA-col, c = statistically significant to col, d = statistically significant to GelMA, e = statistically significant to PVA, f = statistically significant to TCP, g = statistically significant to PCL.

Article Snippet: The cells were stained with anti-human CD 16-APC (10 μL/1 × 10 6 cells) (R&D Systems, Biotechne, USA), FITC mouse anti-human CD66b (20 μL/1 × 10 6 cells) (BD Biosciences, USA) and PE mouse anti-human CD14 (20 μL/1 × 10 6 cells) (BD Biosciences, USA) antibodies for 40 min. After washing cells with 0.5% (w/v) BSA-PBS and centrifugation (300 g for 5 min), cells were analyzed to measure the intensity of CD markers via BD FACSAria™ III cytometer (BD Life Sciences, USA).

Techniques: Activity Assay, Flow Cytometry, Expressing, Staining, Fluorescence, Cell Culture, Lysis