Histamine-Induced Actin Polymerization In Human Eosinophils: An Imaging Approach For Histamine H-4 Receptor

Image-based screening, a new and flexible tool in the drug discovery cascade, is amenable to many different targets. This article describes a particular use of the Cellomics ArrayScan in developing a functional screen for histamine H-4 receptor (H4R) antagonists that have potential utility in inflammatory diseases of the airways such as asthma, with H4R being expressed on a wide variety of immune cells including eosinophils. Exposure to histamine causes eosinophils to migrate from the bloodstream into the tissue where they contribute to inflammation. Migration is manifested through rearrangements of the actin cytoskeleton and phalloidin, a biological peptide, selectively binds F-actin over G-actin and can be used to detect these cytoskeletal changes mediating inflammatory function. A fluorescent conjugate of phalloidin was used to visualize histamine-induced actin polymerization in human cosinophils on the Cellomics ArrayScan. Inhibition of this phenomenon by commercially available histamine receptor antagonists was measured. The selective H4R antagonist JNJ7777120 inhibited histamine-induced actin polymerization in eosinophils most potently. This assay illustrates that this phenomenon is mediated through the H4R and that the image-based format has enhanced screening utility for identifying selective H4R antagonists over traditional flow cytometry methods. (C) 2007 International Society for Analytical Cytology.