Anti human cd14 pe cy7

Anti-human PDPN–PE (clone NZ-1.3, 12-9381-42, 1:100), anti-human CD45–PE-Cy7 (clone HI30, 25-0459-42, 1:100), anti-human CD31–biotin (clone WM59, 13-0319-82, 1:100) and anti-human or mouse ACTA2–eFluor 660 (clone 1A4, 50-9760-82, 1:500) were purchased from eBioscience. Anti-human CD31–PerCP (clone WM59, 303132, 1:50), anti-human CD235a–PE-Cy7 (clone HI264, 349112, 1:100), anti-human CD3–FITC (clone UCHT1, 300440, 1:100), anti-human CD14–PE-Cy7 (clone MSE2, 301813, 1:100), anti-human CD19–APC/Fire 750 (clone HIB19, 302258, 1:100), anti-human CD54 (ICAM1)–BV421 (clone HA58, 353132, 1:100) and anti-human CD34–FITC (clone 581, 343504, 1:100) were purchased from BioLegend. Anti-human CD45–APC-Cy7 (clone 2D1, 560178, 1:100), anti-human CD4–BUV395 (clone SK3, 563550, 1:200), anti-human CD8–BUV805 (clone SK1, 612889, 1:400) and anti-human CD25–BUV563 (clone 2A3, 612918, 1:400) were purchased from BD Biosciences.

Flow cytometric analysis of monocytes was performed as previously described (11 (link)). To this end, cells were stained with the following monoclonal antibodies (BioLegend, Uithoorn, The Netherlands) in PBS supplemented with 0.1% BSA and 0.01% NaN₃: anti-human CD14-PE/Cy7 (clone: HCD14), anti-human CD16-APC/Cy7 (clone: 3G8), anti-human CD163-PE (clone GHI/61), anti-human CD192 (CCR2)-PerCP/Cy5.5 (clone: K036C2), and anti-human CX3CR1-FITC (clone: 2A9-1). Data were acquired on a FACS Canto II device (BD Bioscience) and analyzed using FlowJo® software (Tree Star, Ashland, OR).

For immunophenotyping, nasal cells were dislodged from curettes by pipetting and stained with an antibody cocktail containing anti-human CD3 APC, anti-human CD14 PE-Cy7, anti-human CD66b PE, anti-human CD19 Brilliant Violet 510, and anti-human CD45 Alexa Fluor 700 (all BioLegend, UK). Samples were acquired on an LSR FORTESSA flow cytometer (BD Biosciences, UK) and analyzed using Flowjo v10.5.3 (BD Biosciences, USA).