Expression of pentraxin 3 in equine lungs and neutrophils

Endotoxin-induced diseases cause significant mortality and morbidity in the horse, leading to enormous economic damage to the equine industry. Neutrophils play a critical role in initiating the immune response in the lung. Pattern recognition receptors (PRRs) are programmed to recognize microbial structures unique to pathogens and mount an immune response. Pentraxin 3 (PTX3) is a PRR that is produced at sites of inflammation by many cell types upon stimulation by pro-inflammatory cytokines and agonists, such as endotoxins [also known as lipopolysaccharides (LPS)]. Pentraxin 3 recognizes and binds to many pathogens, activates the complement cascade, and has a role in the clearance of apoptotic and necrotic cells. Recently, PTX3 has been reported to be localized in the specific granules in human and mouse neutrophils, but no reports exist on the in-situ localization of PTX3 in neutrophils and the lungs of horses. Therefore, the objective of this study was to localize the PTX3 protein in normal and LPS-exposed neutrophils and in normal equine lungs. Immunohistochemical data showed PTX3 staining in the bronchial epithelial cells and the vascular endothelium of normal lungs. Immunogold electron microscopy localized PTX3 in the nuclei, cytoplasm, and vesicular organelles of alveolar macrophages, endothelial cells, and pulmonary intravascular macrophages. Immunohistochemical staining for PTX3 in isolated horse neutrophils showed an altered staining pattern in neutrophils stimulated with LPS. These data suggest that neutrophils may be a mobile form of PTX3 that is readily shuttled to the site of inflammation, where it can be released to fine tune a host defense response.