Nuclear Factor-κB in Middle Ear Epithelial Cells: A Methodological Study Using an ELISA

Abstract

Objective—To contribute to the elucidation of the intracellular mechanisms relating to otitis media with effusion (OME) by detecting and quantifying nuclear factor (NF)-κB in middle ear epithelial cells by means of an ELISA. Material and Methods—Using an ELISA technique, NF-κB was identified and quantified in the cytoplasm of cultured epithelial cells from the rabbit middle ear exposed to a well-known inflammatory stimulus: lipopolysaccharide (LPS). Results—NF-κB was measurable with the ELISA in a concentration range of 16–32 ng/μg total cell protein. LPS caused significant enhancement of NF-κB. Furthermore, the NF-κB concentration increased with the duration of LPS exposure. Western blot analysis confirmed the results obtained by ELISA. Conclusion—Further studies are required to determine the role of NF-κB in initiating and regulating the inflammatory response in the middle ear, including the significance of hyperoxia.

• cell cultures
• ELISA
• middle ear epithelium
• nuclear factor-κB

Ovesen T, Barrett TQ. Nuclear factor-κB in middle ear epithelial cells: a methodological study using an ELISA. Acta Otolaryngol 2003; 123: 306–309.

Ovesen T, Barrett TQ. Nuclear factor-κB in middle ear epithelial cells: a methodological study using an ELISA. Acta Otolaryngol 2003; 123: 306–309.