Nuclear factor-kappaB in middle ear epithelial cells: a methodological study using an ELISA.
OBJECTIVE
To contribute to the elucidation of the intracellular mechanisms relating to otitis media with effusion (OME) by detecting and quantifying nuclear factor (NF)-kappaB in middle ear epithelial cells by means of an ELISA.


METHODS
Using an ELISA technique, NF-kappaB 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-kappaB was measurable with the ELISA in a concentration range of 16-32 ng/microg total cell protein. LPS caused significant enhancement of NF-kappaB. Furthermore, the NF-kappaB concentration increased with the duration of LPS exposure. Western blot analysis confirmed the results obtained by ELISA.


CONCLUSIONS
Further studies are required to determine the role of NF-kappaB in initiating and regulating the inflammatory response in the middle ear, including the significance of hyperoxia.
Ovesen
Therese
T
ENT Department, Aarhus University Hospital, Aarhus, Denmark. toves@akh.aaa.dk
Barrett
Thomas Q
TQ
D000818:Animals; D015153:Blotting, Western; D002478:Cells, Cultured; D004432:Ear, Middle / Q000166:cytology*; D004797:Enzyme-Linked Immunosorbent Assay; D004847:Epithelial Cells / Q000378:metabolism*; D016328:NF-kappa B / Q000032:analysis*; D011336:Probability; D011817:Rabbits; D012680:Sensitivity and Specificity
D016328:NF-kappa B