Eotaxin: a Potential Target for Therapy in Chronic Rhinosinusitis Alisha N. West, MD Grand Rounds University of California, Los Angeles December 1, 2010

History Chronic Rhinosinusitis (CRS) is characterized by signs and symptoms of inflammation Stimulated by environmental pathogen Chronic epithelial stimulation  immune response with a subset of patients exhibiting eosinophilia Lack of effective mucociliary clearance Currently, paucity of effective therapies

History Chronic Rhinosinusitis is one of the leading causes of annual visits to the Otolaryngologist 66 million adults in the US reported sinus problems last year $5.8 billion dollars is spent annually on treatment for CRS Symptoms usually recur within three months of therapy

Hypothesis Eotaxin plays a role in CRS inflammation and could serve as a potential target for therapy

Background Eotaxin-1 and 2 implicated in inflammation seen in ABPA and asthma IL-13 and IL-4 up-regulate production of Eotaxin by epithelial cells Eotaxin is a chemotactic factor that attracts eosinophils to the site of inflammation Eosinophils release MBP which has been shown to cause epithelial cell damage in ABPA patients Toxic levels of MBP have been found in the mucus of CRS patients Prior to our current project, the role of eosinophils and eotaxin had not been examined in CRS

Materials and Methods UNC IRB approved protocols Sinonasal mucosa harvested during endoscopic sinus surgery60 subjects Tissue embedded in parrifin and H&E stain performed for EOS Cell culture protocol Cellular disaggregation Plated in serum-free media until confluent, dispersion in trypsin Re-plated on Milicells™ in air-liquid interface media at density of 250,000 cells/Millicell™ After confluent monolayeraspirate apical media Washed and re-fed basally for 21 days until ciliated and mucus production 4 cell cultures/specimen Apical challenge with Pseudomonas, Staph, and TSB Basal Media collected, washed and replaced daily 0-96 hours

Materials and Methods Luminex 22-plex battery of Cytokines IL-1beta IL-2 IL-4 IL-5 IL-6 IL-10 IL-12p70 TNF-alpha IFN-gamma G-CSF IP-10 GM-CSF IL-9 IL-13 KC MCP-1 MIP-1a RANTES IL-1a IL-7 IL-15 IL-17

Materials and Methods Alveolar Lavage has demonstrated eosinophilia in ABPA and asthma patients as well as animal models Endosinus lavage with evaluation of eosinophilia in CRS patients and normal volunteers Complete cell count and Eosinophil count performed with cytospin

Tissue Harvest

Results

H&E Stain 10x CRS Control

Eosinophilia CRS Control

Eosinophilia in Endosinus Mucosa

Eotaxin ELISA

Luminex Results

Conclusions There is significant Eosinophilia in CRS endosinus epithelium when compared to normal controls CRS epithelium produces a higher concentration of eotaxin at baseline and after an infectious challenge when compared to controls Eotaxin should be investigated as a potential target for therapy in CRS

References

Thank You