Identification of CD Marker Expression and Neutrophil Surface Marker Changes in Health and Disease using High-throughput Screening Flow Cytometry

Neutrophil hyperactivation can contribute to tissue damage in inflammatory diseases. Although many cell-surface proteins are known to be expressed on neutrophils there is no comprehensive study of the surface-markers that can be used to phenotype neutrophils. Neutrophils subpopulations isolated (blood and oral rinses) from healthy and chronic-periodontitis patients were screened against a panel of 374 known Cluster of Differentiation (CD) antibodies to identify cell-surface markers specific to neutrophils. This screen identified CD11b+, CD16b+, and CD66c+ as markers that are always expressed on neutrophils. Cell-sorting with an antibody against CD11b/CD16b/CD66c allowed for the enrichment of mature neutrophils, yielding populations of up to 99%, confirming the validity of these markers when isolating neutrophils. These findings provide a simple method for isolating neutrophils from humans, and thereby establish a validated method that allows for the accurate identification of neutrophils. This knowledge will be crucial for identifying neutrophil subtypes associated with neutrophil-mediated inflammatory diseases.M.Sc