Detection of Chemokines in Nasal Secretion Using Filter Paper

Abstract

It is widely assumed that cytokines and chemokines play a significant role in the pathogenesis of inflammatory diseases such as allergic rhinitis. Much of the data supporting this concept has been generated under in vitro experimental conditions; a definitive proof of the involvement of these factors in inflammation would require the recovery of factors from inflammatory sites. Although washing and lavage of inflammatory sites is routinely done to detect factors at the site, relatively high concentrations of factors are only measurable by conventional methods. We attempted to detect chemokines in nasal secretion by filter paper and to determine possible mechanisms of chemokines in the development of allergic rhinitis. Small filter strips (7×30mm) were placed on nasal turbinates for 1min. After being air-dried strips were cut into small pieces and factors were extracted by rocking for 12h at 4°C. The eluate was collected and assayed for eotaxin and RANTES by ELISA. The eluate from normal control subjects contained detectable levels of both eotaxin and RANTES. These factor levels were significantly increased in patients with pollinosis. Levels of eotaxin, but not of RANTES, correlate closely with the presence of eosinophils in nasal mucosa. These results suggest that eotaxin is an important factor in nasal mucosal eosinophilia in pollinosis.