Multiplex Biomarker Profiling in BALF

MMP9 (EMD MILLIPLEX MAP MMP Magnetic Bead Panels, EMD Millipore, Billerica, MA) and cytokines (IL-13, RANTES, CXCL2, TNF-α, MCP-1, IL-1β, IL-17, IL-10, CXCL1, G-CSF, IL-6 and human IL-8; Bio-Rad Cytokines Magnetic Bead Panels) were measured in BALF and cell culture supernatants using a bead assay (MILLIPLEX MAP MMP Magnetic Bead Panels, EMD Millipore, Billerica, MA) with the Bio-Rad Bio-Plex 200 system (Bio-Rad, Hercules, CA) or with commercially available ELISAs (CXCL5 and SCF; R&D Systems). BALF gelatinase activity was determined by gelatin zymography [13 (link)]. Densitometry was performed on MMP9 positive bands and represented densitometry units (DU) measured by pixel intensity of bands, using Bio-Rad Laboratories Image Lab software (version 4.0, build 16).