Immunohistochemical analysis of the activation of NF-?B and expression of associated cytokines and adhesion molecules in human models of allergic inflammation

✍ Scribed by Wilson, Susan J.; Leone, Barbara A.; Anderson, Donald; Manning, Anthony; Holgate, Stephen T.

• Publisher: John Wiley and Sons
• Year: 1999
• Tongue: English
• Weight: 407 KB
• Volume: 189
• Category: Article
• ISSN: 0022-3417
• DOI: 10.1002/(sici)1096-9896(199910)189:2<265::aid-path415>3.0.co;2-#

✦ Synopsis

To investigate the role of NF-B in regulating allergic inflammation, a monoclonal antibody directed to the activated form of NF-B has been developed and immunohistochemistry has been employed to study the pro-inflammatory transcriptive function of NF-B and the adhesion molecules and cytokines that it regulates. Human umbilical vein endothelial cells (HUVECs) exposed to physiological levels of TNF demonstrated dose-and time-dependent cytoplasmic and nuclear activation of NF-B, followed by up-regulation of ICAM-1. This was suppressed by the selective inhibitors of NF-B activation, calpain and gliotoxin. Using monoclonal antibodies directed to NF-B and associated cytokines and adhesion molecules, immunohistochemistry was applied to bronchial explants stimulated ex vivo with TNF , and to nasal polyp tissue, embedded in glycol methacrylate. Stimulation of the bronchial explants increased expression of NF-B, IL-8, and GM-CSF in the epithelium and endothelium and ICAM-1 in the endothelium. In nasal polyp, expression of NF-B was in the epithelium, the endothelium and in submucosal mast cells, eosinophils, T and B lymphocytes, and macrophages. Thus, immunohistochemistry can be used to determine the cellular provenance of NF-B and its activation status in single cell and complex tissue systems, in parallel with appropriate inflammatory markers.